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We investigated the effect of an ABCE1-silencing gene on the proliferation, invasion, and migration of human thyroid carcinoma SW579 cells. We designed and synthesized targeted ABCE1-siRNA sequences and a negative control sequence (NC-siRNA), and transfected them into human thyroid cancer SW579 cells by electrotransfer to obtain ABCE1-SW579 and NC-siRNA-SW579 cells (siRNA is small interfering RNA). Through reverse transcription polymerase chain reaction and western blotting analysis, ABCE1 mRNA and protein expression levels in the electrotransferred cells were detected, and flow cytometry was used to detect cell cycle and apoptosis. The cell counting kit-8 (CCK-8) proliferation assay, the scratch healing assay, and the cell invasion assay were used to measure cell proliferation, migration, and invasion capabilities, respectively. Compared with NC-siRNA-SW579 and Ctrl-SW579 groups, ABCE1 mRNA and protein expression levels in the ABCE1-SW579 cells were significantly reduced. The growth rate of ABCE1-SW579 cells was significantly inhibited, the cell cycle was arrested in the G0/G1 phase, and the number of cells in the S phase was reduced. Compared with the Ctrl-SW579 group, the cell apoptosis rate in the ABCE1-SW579 group was significantly higher (P < 0.01), and proliferation, migration, and invasion were significantly reduced (P < 0.05). Expression of the ABCE1-silencing gene, transfected by electrotransfer, could inhibit the proliferation, invasion, and migration of thyroid cancer cells.

OBJECTIVE: The presence of obesity and increases in body mass are important risk factors for cardiovascular disease and diabetes. This study examined the effects of olanzapine, risperidone, and haloperidol on weight, body mass index (BMI), and development of obesity in a drug-naive population compared with a matched healthy control group.METHOD: Consecutive patients during the period from June through October 2006 with DSM-IV schizophrenia at our referral psychiatric hospital were recruited for an extensive prospective study that included anthropometric measures of weight, waist circumference, waist-hip ratio, and BMI. Subjects were randomly assigned to receive haloperidol, olanzapine, or risperidone and compared with a matched healthy control group. The prevalence of obesity, which was the main outcome measure, was assessed on the basis of 2 criteria: revised World Health Organization (WHO) definition for Asians and criteria of the International Diabetes Federation (IDF). Inclusions started in June 2006, and patients were followed for a period of 6 weeks.RESULTS: The analysis of 66 patients showed a prevalence of overweight (WHO criteria) at 22.7% and obesity at 31.8% (IDF criteria). The prevalence of obesity (IDF criteria) in our patients is over 30 times as high as that of the matched healthy control group (p < .001). Subjects in the olanzapine group had the greatest weight gain at 5.1 kg, followed by risperidone at 4.1 kg and haloperidol at 2.8 kg.CONCLUSIONS: Obesity is highly prevalent among patients treated with atypical antipsychotics for schizophrenia. Assessment and monitoring of obesity along with preventive and curative measures should be part of the clinical management of patients treated with antipsychotics.TRIAL REGISTRATION: ClinicalTrials.gov, NCT00534183, www.clinicaltrials.gov.

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases. Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema. As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor. In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix. The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid. Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion. It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used. The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

BACKGROUND AND OBJECTIVES: The objectives of this study were to investigate the effects of cinnamon supplementation on glycemic indices, serum lipids and adiponectin in patients with polycystic ovary syndrome (PCOS).METHODS AND STUDY DESIGN: This double-blind randomized controlled clinical trial was conducted on 84 overweight or obese PCOS patients. Subjects in cinnamon (n=42) and placebo (n=42) groups were given 3 cinnamon capsules (each one contained 500 mg cinnamon) or placebo daily for 8 weeks. Fasting blood samples, anthropometric measurements and dietary intake data were collected at the baseline and at the end of the trial. Data were analyzed by independent t test, paired t test and analysis of covariance.RESULTS: Cinnamon significantly decreased serum fasting blood glucose, insulin, homeostatic model assessment for insulin resistance, total cholesterol and low-density lipoprotein cholesterol and weight and increased high-density lipoprotein cholesterol compared with placebo (all p<0.05). Serum triglyceride and body mass index significantly decreased in the cinnamon group, in comparison with baseline values (p=0.001 and p=0.002, respectively). No significant changes were seen in serum adiponectin in either group.CONCLUSIONS: Short term supplementation of cinnamon had some favourable effects on metabolic risk factors of women with PCOS and may be useful in management of PCOS complications.

Central nervous system (CNS) has a completely separate immune system that communicates with the neurons by small molecules called cytokines. Cytokines are involved in many crucial processes in neuron including cell metabolism and neurotransmitter synthesis. It has been reported that cytokine imbalance is involved in the progression of many CNS diseases such as neuropsychiatric disorders (depression, schizophrenia, autism, and bipolar disorder) and neurodegenerative disorders (Parkinson's and Alzheimer's disease). Here, the effects of diclofenac, different antidepressants (sertraline, venlafaxine, and fluvoxamine), and vitamin B₆ (pyridoxine) on IL-10 and tumor necrosis factor-á (TNF-á) change with and without immune challenges with lipopolysaccharide (LPS) were investigated in in vitro culture of astrocytes from 2-day-old Swiss-Albino mice. Diclofenac and Sertraline significantly (p < 0.05) improves anti-inflammatory cytokine (IL-10) while suppress (p < 0.05) LPS-induced elevated level of pro-inflammatory mediators (TNF-á) in astrocyte culture. Pyridoxine was not able to reduce (p > 0.05) TNF-á in the astrocyte culture. Antidepressant (sertraline) showed positive effects (increased IL-10 and reduced TNF-á level) possibly through the suppression of Th1 lymphocytes and monocytes and stimulation of Th2 lymphocytes and monocytes/macrophages. NSAID (diclofenac) showed positive immune regulation effect possibly through the inhibition of cyclo-oxygenase enzyme. Based on these findings, it may conclude that, diclofenac and antidepressants (sertraline) may positively contribute in the cytokine production in astrocyte cell culture.

BACKGROUND: Glomerulomegaly, the abnormal enlargement of glomeruli, has been related to an increased risk of glomerulosclerosis, but the degree of enlargement that constitutes glomerulomegaly has not been defined.METHODS: The principal stereological methods for estimating glomerular volume are [1] the disector/Cavalieri method that is considered the 'gold standard' for measuring individual glomerular volume (IV(glom)) and [2] the disector/fractionator technique that estimates average glomerular volume (V(glom)) together with total glomerular number (N(glom)) for the entire kidney. The two methods produce different estimates with V(glom) consistently exceeding IV(glom). This study compares glomerular volumes obtained by the two methods in autopsy kidneys of 39 African American and 34 US white adult males, and correlates the values with N(glom), body mass index (BMI), hypertension, glomerulosclerosis and race, factors known or thought to influence glomerular volume.RESULTS: For the smallest glomeruli, V(glom) was 25% larger than IV(glom) with the difference increasing to over 50% for kidneys with the largest glomeruli. Both V(glom) and IV(glom) showed significant inverse correlations with N(glom) and significant direct correlations with BMI and hypertension. African Americans had larger IV(glom) and V(glom) than whites, but only IV(glom) was significant. The 90th percentile for IV(glom) was 6.81 ìm(3) ? 10(6) and 13.10 ìm(3) ? 10(6) for V(glom), but larger glomerular size did not separate hypertensive from non-hypertensive subjects nor did it show any significant relationship to glomerulosclerosis. While V(glom) overestimated glomerular size compared with IV(glom), both measurements demonstrated similar relationships to factors influencing glomerular volume.CONCLUSIONS: With neither method could glomerulomegaly, the abnormal enlargement of glomerular size predisposing to glomerulosclerosis, be determined.

The role of U. urealyticum in spontaneous and recurrent spontaneous abortion was studied in 633 women. Cervical colonization with U. urealyticum was found in 42.6% of 310 normal pregnant women, in 41.6% of 84 patients who underwent induced abortion, in 41.5% of 41 normal fertile patients, in 53.3% of 122 patients with spontaneous abortion and in 64.5% of 76 patients with recurrent spontaneous abortion. The cervical isolation rate was significantly higher in patients with spontaneous abortion (p less than 0.05) and recurrent spontaneous abortion (p less than 0.005) than in normal pregnant women. Endometrial colonization was more frequent in patients with recurrent spontaneous abortion (27.6%) than in normal fertile women (9.7%) (p less than 0.05). Moreover, in 6 patients with intact membranes and uncontrollable preterm labor resulting in fetal loss (all between the 20th and 28th week of gestation) U. urealyticum was isolated in 5 of them from the cervix, in 4 patients from the placenta and in 2 out of 4 from the amniotic fluid. Histological examination of the placenta showed signs of chorioamnionitis in 5 patients. From this study we conclude that although U. urealyticum is a common inhabitant of the lower genital tract, it may play a role in the etiology of spontaneous abortion and uncontrollable preterm labor.

Artificial neural network (ANN) analysis was used to predict the permeability of selected compounds through Caco-2 cell monolayers. Previously reported models, which were shown to be useful in the prediction of permeability values, use many structural parameters. More complex equations have also been proposed using both linear and non-linear relationships, including ANN analysis and various structural parameters. But proposed models still need to be developed using different neuron patterns for more precise predictions and a better understanding of which factors affect the permeation. To develop a simple and useful model or method for easy prediction is also a general need. Permeability coefficients (log kp) were obtained from various literature sources. Some structural parameters were calculated using computer programs. Multiple linear regression analysis (MLRA) was used to predict Caco-2 cell permeability for the set of 50 compounds (r2=0.403). A successful ANN model was developed, and the ANN produced log kp values that correlated well with the experimental ones (r2=0.952). The permeability of a compound, famotidine, which has not previously been studied, through the Caco-2 cell monolayer was investigated, and its permeability coefficient determined. It was then possible to compare the experimental data with that predicted using the trained ANN with previously determined Caco-2 cell permeability values and structural parameters of compounds. The model was also tested using literature values. The developed and described ANN model in this publication does not require any experimental parameters; it could potentially provide useful and precise prediction of permeability for new drugs or other penetrants.

In May 2015, a child with absence of most of the five fingers with scar formation after healing of a left hand burn wound hospitalized in our burn ward. According to the free online design program for making artificial limbs using three-dimensional printing technology on the internet, a utility artificial hand, most of which made of plastic parts, was designed for the child and printed by a three-dimensional printer. The child was instructed to wear and use the utility artificial hand, including driving the finger part of the utility artificial hand to make a grasping action by flexing the wrist joint. On the first day of using the utility artificial hand, the time the right hand and the utility artificial hand took to finish the Nine-Hole Peg Test (NHPT) was 24 and 325 s, respectively. After training, the child could grab some light and rough objects. After 3 months of follow-up, the child could use the utility artificial hand to cooperate with the upper limb of the healthy side to make the movements of picking up the basketball and keeping the balance of body on the bicycle. The time the right hand and the utility artificial hand took to finish NHPT was 21 and 193 s, respectively. The time the utility artificial hand took increased by 40.6% compared with the initial period. By assembling the three-dimensionally printed utility artificial hand, the partial appearance image of the child was restored, and some of the hand functions were compensated, which improved the self-care ability of the child in daily life and was beneficial to his physical and mental development.

Pentafraction (PF), a solution of biodegradable hydroxyethyl starch macromolecules with molecular weights of 10 to 100 x 10(4) daltons, has been shown to minimize tissue edema by sealing interendothelial clefts at the capillary level. The effect of PF on ischemia-reperfusion-induced compartment syndrome was studied. Ten rabbits underwent bilateral femoral artery occlusion following ligation of branches from the terminal aorta to the popliteal artery. After 7 hours of ischemia, reperfusion was established with heparinized polyethylene shunts. Experimental animals (n = 5) received PF and control animals (n = 5) received normal saline (NS) as an intravenous infusion (30 mL/kg) for 1 hour, beginning 10 minutes after shunt placement. During reperfusion, anterior compartment pressure was continuously monitored in the left lower extremity. To quantitate oxidative metabolism, triphenyltetrazolium chloride (TTC) reduction (micrograms of TTC per milligram of protein) of tibialis anterior muscle from the right lower extremity was measured at femoral artery occlusion, 7 hours of ischemia, and 2 hours of reperfusion. In the NS group, anterior compartment pressure significantly increased from the end of the ischemic interval, 10.8 +/- 4.14 to 36.4 +/- 9.9 mmHg and 44.6 +/- 15.4 mmHg, after 1 and 2 hours of reperfusion (p < 0.007) compared with the PF group, which did not change significantly, 10.6 +/- 2.6 to 11.4 +/- 12.9 mmHg and 7.4 +/- 2.8 mmHg, after 1 and 2 hours of reperfusion (p < 0.67).(ABSTRACT TRUNCATED AT 250 WORDS)

We have previously isolated rabbit liver microsomal metalloendopeptidase (MEP) as a candidate for the processing enzyme of vitamin K-dependent plasma proteins. A cDNA coding for MEP has revealed that it is structurally related to metalloendopeptidase-24.15, which catalyzes the proteolytic processing of several bioactive peptides. In this study we examined the tissue distribution and subcellular localization of MEP by light and electron microscopic immunohistochemical methods, in addition to Northern blot analysis. Chicken polyclonal antibodies were raised by using synthetic peptides AG1 (Met31-Asn46) and AG3 (Asp537-Gly551) derived from the sequence of MEP. Both anti-AG1 and anti-AG3 antibodies reacted specifically with MEP, as judged by Western blotting and immunohistochemical methods. Both antibodies gave an identical staining distribution, which was localized on the luminal cell surfaces and in the cytoplasm of the following organs: liver, brain, lungs, kidneys, esophagus, stomach, duodenum, pancreas, placenta, epididymis, uterus, ovary, and oviduct. Northern blot analysis revealed that the expression of MEP mRNA is similar to its immunohistochemical distribution except in the heart. These results suggest that MEP may participate more closely in a degradation role in peptide metabolism in various tissues than in a processing role of the proprotein, like metalloendopeptidase-24.15.

Elevated serum total homocysteine (tHcy) is an established risk factor for cardiovascular disease. Its role in dementia is still controversial, and no study has examined the role of midlife tHcy, or reports longer than 8 years of follow-up. We examined the relation between midlife tHcy and late-life dementia in women followed for 35 years. The Prospective Population Study of Women in Gothenburg began in 1968-1969, comprising a representative population of women aged 38-60 years. Four extensive follow-ups were conducted by 2003. Serum samples from 1968 to 1969 were analysed for tHcy in 1368 women. In total, 151 women developed dementia. The highest tHcy tertile was related to a hazard ratio of 1.7 (95% CI 1.1-2.6) for developing any dementia, 2.1 (95% CI 1.2-3.7, n=100) for AD and 2.4 (95% CI 1.3-4.7, n=68) for AD without cerebrovascular disease. Kaplan-Meier plots showed divergence with respect to dementia after 22 years of follow-up. In conclusion, high homocysteine in midlife is an independent risk factor for the development of late-life Alzheimer dementia in women.

Since the seventies, the practice of drug smuggling in the form of body packing has increased in the Western world. The goal of our study was to present an algorithm for the safe management of intracorporal drug transport based on clinical experience and current evidence. The retrospective study, conducted over the past four years in our hospital prison, analyzes and discusses the diagnostic and therapeutic concepts. Thirty-four patients hospitalized 37 times in a 48-month period were included. In 28 patients drug packages were identified. Only two patients suffered from serious complications. The study demonstrates that following a specifically designed management algorithm based on clinical experience and principles of evidence-based medicine can optimize risk management, improve quality assurance and patient safety.

BACKGROUND AND AIM: Once thought to be uncommon in Asia, coeliac disease (CD) is now being increasingly recognized in Asia–Pacific region. In many Asian nations, CD is still considered to be either nonexistent or very rare. In recognition of such heterogeneity of knowledge and awareness, the World Gastroenterology Organization and the Asian Pacific Association of Gastroenterology commissioned a working party to address the key issues in emergence of CD in Asia.METHODS: A working group consisting of members from Asia–Pacific region, Europe, North America, and South America reviewed relevant existing literature with focus on those issues specific to Asia–Pacific region both in terms of what exists and what needs to be done.RESULTS: The working group identified the gaps in epidemiology, diagnosis, and management of CD in Asian–Pacific region and recommended the following: to establish prevalence of CD across region, increase in awareness about CD among physicians and patients, and recognition of atypical manifestations of CD. The challenges such as variability in performance of serological tests, lack of population-specific cut-offs values for a positive test, need for expert dietitians for proper counseling and supervision of patients, need for gluten-free infrastructure in food supply and creation of patient advocacy organizations were also emphasized.CONCLUSIONS: Although absolute number of patients with CD at present is not very large, this number is expected to increase over the next few years or decades. It is thus appropriate that medical community across the Asia–Pacific region define extent of problem and get prepared to handle impending epidemic of CD.

The authors studied growth peculiarities of the ascitic hepatoma 22A at the terminal stage of development. As shown by the autoradiography method, some of the cells could be at the G1-period or at the R1-period for 2 or even 4 days; the average duration of the G2-period constituted 16 hours. 55--60% of the cells were at the much prolonged G1- and R1-period, 7%--at the S-period, and 9%--at the G2-period. The rest 25--30% of cells apparently leave the mitotic cycle irreversibly since they take no part in proliferation after the division stimulation.

Human intestinal lamina propria T lymphocytes (LPL-T) physiologically exhibit minimal proliferation in response to antigen receptor stimulation in vitro. This is thought to occur as a consequence of regulatory influences which are exerted by the mucosal microenvironment. The present study is aimed at investigating whether proliferative responses of intestinal LPL-T to antigen receptor stimulation are altered in patients with inflammatory bowel disease. Accordingly, proliferative responses of LPL-T in patients with Crohn's disease and ulcerative colitis to stimulation with CD3 MoAb plus IL-2 were examined and compared with controls. In addition, T cell receptor (TCR) repertoires of LPL-T and peripheral blood T lymphocytes were determined by indirect immunofluorescence using a panel of 11 TCR V beta specific antibodies. In most patients with inflammatory bowel disease, LPL-T showed enhanced proliferation to antigen receptor stimulation compared with controls. Moreover, perhaps as a consequence, an enhanced frequency of in vivo preactivated T cells was seen as judged from an increased spontaneous proliferative response to low concentrations of exogenous IL-2. LPL-T and peripheral blood T lymphocytes exhibited similar percentages of TCR V beta gene usage both in controls and in patients. In summary, polyclonal activation of LPL-T due to impairment of local adjustment, i.e. insufficient down-regulation of TCR/CD3-dependent signalling processes, may contribute to the pathogenesis of inflammatory bowel disease.

Many robust tests have been proposed in the literature to compare two hazard rate functions, however, very few of them can be used in cases when there are multiple hazard rate functions to be compared. In this article, we propose an approach for detecting the difference among multiple hazard rate functions. Through a simulation study and a real-data application, we show that the new method is robust and powerful in many situations, compared with some commonly used tests.

The major glycoproteins of bovine gammaherpesvirus 4 (BoHV-4) are gB, gH, gM, gL, and gp180 with gB, gH, and gp180 being the most glycosylated. These glycoproteins participate in cell binding while some act as neutralization targets. Glycosylation of these envelope proteins may be involved in virion protection against neutralization by antibodies. In infected cattle, BoHV-4 induces an immune response characterized by low neutralizing antibody levels or an absence of such antibodies. Therefore, virus seroneutralization in vitro cannot always be easily demonstrated. The aim of this study was to evaluate the neutralizing capacity of 2 Argentine BoHV-4 strains and to associate those findings with the gene expression profiles of the major envelope glycoproteins. Expression of genes coding for the envelope glycoproteins occurred earlier in cells infected with isolate 10/154 than in cells infected with strain 07/435, demonstrating a distinct difference between the strains. Differences in serological response can be attributed to differences in the expression of antigenic proteins or to post-translational modifications that mask neutralizing epitopes. Strain 07/435 induced significantly high titers of neutralizing antibodies in several animal species in addition to bovines. The most relevant serological differences were observed in adult animals. This is the first comprehensive analysis of the expression kinetics of genes coding for BoHV-4 glycoproteins in 2 Argentine strains (genotypes 1 and 2). The results further elucidate the BoHV-4 life cycle and may also help determine the genetic variability of the strains circulating in Argentina.

Host status for Mexican fruit fly (Anastrepha ludens (Loew)) was examined under laboratory conditions in cage infested Eureka and Meyer lemons. Our approach was to allow females to oviposit on the two cultivars in separate laboratory cages with aluminum foil covering to restrict the areas where females had access to fruit surface. Fruit of each cultivar were placed in covered trays for incubations and at approximately weekly intervals, fruit were removed, dissected, and live and dead eggs and larvae tabulated in each tissue of the fruit. Infestation and survival were tabulated and analyzed for the effects of harvest date, fruit color and brix indices, postoviposition period, and cultivar. Infestation rate, determined by counts of total eggs and larvae was significantly higher in Meyer lemons. In both cultivars, females deposited eggs into both albedo and pulp tissue but not into flavedo. Both cultivars showed high resistance (> 90% mortality) to egg and first instars survival in albedo and pulp. Second and third instars surviving in the pulp had high survival rates (> 60%) in both cultivars in fruit dissected at weeks 2-4 after infestation. Total adults produced were slightly higher, and total second and third stage larvae were also higher for Meyer lemons. Numbers of adults and total second and third stage larvae increased in Eureka lemons in more mature fruit, but the higher numbers in Meyer lemons were not associated with fruit maturity, at time of infestation. Numbers of second and third stage larvae were significantly correlated with some fruit color indices in Eureka but not in Meyer lemons. Application of these results to quarantine risk analysis is discussed.

The effect of a proteinase inhibitor extracted from potatoes (POT II) which increases CCK release, on food intake was examined in 11 lean subjects. They received 1.5 g POT II in a high-protein soup vehicle (70 kcal), the soup vehicle alone, or a no-soup control five minutes before being presented with a lunchtime test meal, according to a double-blind, within-subjects design. Consuming the soup alone led to a nonsignificant 3% reduction in energy intake. The addition of 1.5 g POT II to the soup significantly reduced energy intake by a further 17.5%. Premeal ratings of motivation to eat and food preferences did not predict the reduction in energy intake by the proteinase inhibitor. These findings suggest that endogenous CCK may be important in the control of food intake and that proteinase inhibition may have therapeutic potential for reducing food intake.

Fertility rates of donkey semen in jennies are lower compared to mares. The aims of this study were to evaluate different sperm cryopreservation methods and insemination strategies to improve the fertility of donkey semen in jennies. Three experiments were performed: (1) the comparison of two freezing methods of donkey semen (conventional method and automated method); (2) the determination of a suitable insemination dose of fresh donkey semen for jennies and mares; and (3) the influence of the semen deposition site on fertility of jennies inseminated with frozen donkey semen. For experiment 1, no differences were observed in total motility, angular velocity, curvilinear velocity, straight-line velocity, and plasma membrane integrity between samples frozen with the conventional (Styrofoam box) and the automated method (TK 4000C). However, the automated method provided higher values of progressive motility and rapid cells in frozen-thawed samples in comparison with the conventional method (P < 0.05). For experiment 2, mares were bred using 500 ? 10(6) fresh sperm (M); and jennies using 1 ? 10(9) (J1) or 500 ? 10(6) fresh sperm (J5). Pregnancy rates in M, J1, and J5 were 93% (14/15), 73% (11/15), and 40% (6/15), respectively. When using different insemination doses, 500 ? 10(6) or 1 ? 10(9) sperm, no significant difference was observed in pregnancy rates of mares (M, 14/15) and jennies (J1, 11/15). Furthermore, there was no significant difference between the two insemination doses in jennies. However, with an insemination dose of 500 ? 10(6) fresh sperm, the pregnancy rates were significantly higher in mares (M, 14/15) than in jennies (J5, 6/15; P < 0.05). For experiment 3, the inseminations were carried out in the uterine body (UB) or in the uterine horn of jennies with frozen-thawed donkey semen. No pregnancies were achieved with inseminations performed in the UB (0/12). The pregnancy rate for uterine horn group was 28.26% (13/46) and thus significantly higher than the UB group (0%; 0/12; P < 0.05). In conclusion, the automated method showed higher values on progressive motility and rapid cells parameters compared to the conventional method and can be used as an alternative for freezing donkey semen. The increase in the number of sperm cells per insemination dose using fresh donkey semen improved the fertility rates in jennies. The deep horn inseminations using frozen-thawed donkey semen increased the pregnancy rate in jennies, and the multiple inseminations may be an option to improve the fertility rates of donkey semen in jennies.

In his 1756 text, Observations pratiques sur les maladies de l'ur?thre et sur plusiers faits convulsifs, Nicolas Andr? coined the term "tic douloureux." He believed that this pain originated from compression of facial sensory peripheral nerves. Using scientific observation and experimentation to confirm this hypothesis, he reproduced the tic pain and treated it by using careful efforts to remove adhesions from the nerve with a caustic solution of mercury water. Believing that recurrence of the pain was a result of early closure of the wound, with recompression of the nerve being the direct cause, Andr? prevented recompression by ensuring open wound drainage. Andr?'s surgical technique of using cauterizing stones ensured that there was minimal blood loss and little danger of rebleeding and recompression of the nerve by an accumulated blood clot. His case reports include lengthy follow-up periods that documented the benefits of his procedures, which were confirmed by testimonials from uninvolved colleagues. Although remembered for the two words, "tic douloureux," Nicolas Andr? has long been ignored for his prescient treatment and scientific analysis of a disease for which the modern standard of care has only been defined during the last generation.

Effort thrombosis of the axillary-subclavian vein (Paget-Schroetter syndrome) develops usually secondary to heavy arm exertion. An underlying chronic venous compressive anomaly at the thoracic outlet or intimal damage of the axillary vein following forceful hyperabduction, external rotation of the shoulder joint has been proposed to explain the pathophysiology of this thrombosis. This condition is usually not attributed to an underlying hypercoagulability such as deficiency of natural coagulation inhibitors. Here, the authors present a case with thrombosis of the axillary-subclavian vein following an effort, with factor V Leiden and prothrombin 20210A mutations. Both factor V Leiden and the genetic variant in the prothrombin gene have been shown to confer an increased risk for venous thrombosis. Although rare, effort thrombosis may develop in a patient with hereditary thrombophilia, so laboratory evaluation should include the common causes of thrombosis.

Endothelium exposed to fluid shear stress (FSS) undergoes cell shape change, alignment and microfilament network remodeling in the direction of flow by an unknown mechanism. In this study we explore the role of tyrosine kinase (TK) activity, intracellular calcium ([Ca2+]i), mechanosensitive channels and cytoskeleton in the mechanism of cell shape change and actin stress fiber induction in bovine aortic endothelium (BAE). We report that FSS induces beta-actin mRNA in a time- and magnitude-dependent fashion. Treatment with quin2-AM to chelate intracellular calcium release and herbimycin A to inhibit TK activity abolished BAE shape change and actin stress fiber induction by FSS, while inhibition of protein kinase C with chelerythrine had no effect. Altering intermediate filament structure with acrylamide did not affect alignment or F-actin induction by FSS. Examining the role of the BAE cytoskeleton revealed a critical role for microtubules (MT). MT disruption with nocodazole blocked both FSS-induced morphological change and actin stress fiber induction. In contrast, MT hyperpolymerization with taxol attenuated the cell shape change but did not prevent actin stress fiber induction under flow. Mechanosensitive channels were found not to be involved in the FSS-induced shape change. Blocking the shear-activated current (IK.S) with barium and the stretch-activated cation channels (ISA) with gadolinium had no effect on the shear-induced changes in morphology and cytoskeleton. In summary, FSS has a profound effect on endothelial shape and F-actin network by a mechanism which depends on TK activity, intracellular calcium, and an intact microtubule network, but is independent of protein kinase C, intermediate filaments and shear- and stretch-activated mechanosensitive channels.

Hair follicle morphogenesis, a complex process requiring interaction between epithelia-derived keratinocytes and the underlying mesenchyme, is an attractive model system to study organ development and tissue-specific signaling. Although hair follicle development is genetically tractable, fast and reproducible analysis of factors essential for this process remains a challenge. Here we describe a procedure to generate targeted overexpression or shRNA-mediated knockdown of factors using lentivirus in a tissue-specific manner. Using a modified version of a hair regeneration model, we can achieve robust gain- or loss-of-function analysis in primary mouse keratinocytes or dermal cells to facilitate study of epithelial-mesenchymal signaling pathways that lead to hair follicle morphogenesis. We describe how to isolate fresh primary mouse keratinocytes and dermal cells, which contain dermal papilla cells and their precursors, deliver lentivirus containing either shRNA or cDNA to one of the cell populations, and combine the cells to generate fully formed hair follicles on the backs of nude mice. This approach allows analysis of tissue-specific factors required to generate hair follicles within three weeks and provides a fast and convenient companion to existing genetic models.

Three different pathophysiological mechanisms are probably responsible for hereditary pseudohypoparathyroidism: 1) a defect at the prereceptor-level, 2) a defective membrane N-protein accounting for diminished second messenger production, and 3) a defect in the cytosolic response to the hormone. In a cooperative, study 24 patients (mean age, 13 yr; range, 3-23 yr, 8 girls, 16 boys) receiving vitamin D metabolites (5,000-80,000 U/day) were examined and compared to a control group of 36 normal children. Immunoreactive N-terminal PTH (N-PTH), mid-C-regional PTH (mid-C-PTH), intact PTH and bio-PTH, vitamin D metabolites, and serum calcium and phosphate, alkaline phosphatase activity, and the N-protein activity of erythrocyte membranes were measured in each subject. By clinical and biochemical criteria three groups were differentiated. Eight patients had the completely expressed features of Albright's Hereditary Osteodystrophy (AHO+), including brachydactyly and/or sc calcifications, and increased N-PTH, mid-C-PTH, and alkaline phosphatase activity. Bio-PTH, intact PTH, and N-protein were normal. Nine additional patients with complete (AHO+) had elevated levels of bio-PTH, N-PTH, and mid-C PTH, normal hydroxylation of vitamin D, but decreased N-protein activity. Seven patients with pseudohypoparathyroidism had no features of AHO (AHO-), no increase of urinary cAMP excretion after exogenous PTH, normal PTH peptide levels and N-protein activity, but elevated 25-hydroxyvitamin D and decreased 1,25-dihydroxyvitamin D concentrations. In conclusion, we identified three subpopulations of PsHP: group a had a dissociation of N-PTH and bio-PTH suggesting a defective N-PTH causing renal resistance, whereas their bones respond to PTH. Group b had defective N-protein causing generalized PTH resistance. Group c was characterized by high 25-hydroxyvitamin D and relatively low 1,25-dihydroxyvitamin D levels, thus providing evidence for a defect in the cytosolic interaction of the two different second messengers for PTH, cAMP, and calcium.

Energy and nutrient digestibility of solvent-extracted canola meal (CM) is limited in pigs by its relatively high fiber content. The seed hull, which greatly contributes to the fiber content of CM, is denser than the oil-free cotyledon. By utilizing streams of air, air classification partially separates these seed components on the basis of their different sizes and densities to produce a low-fiber, light-particle fraction and a high-fiber, heavy-particle fraction. Compared with parent CM, ADF and NDF were reduced by 31.9% and 29.5% in the light-particle fraction and were enriched by 16.5% and 9.0% in the heavy-particle fraction (DM basis), respectively. Particle size was 638, 18.9, and 76.1 µm for the parent CM and light- and heavy-particle fractions, respectively. To determine the nutrient digestibility of CM and their air-classified fractions, Brassica napus and B. juncea CM and their 2 air-classified fractions were evaluated in a 2 ? 3 factorial arrangement together with a basal diet and an N-free diet. The experiment was conducted as an 8 ? 8 Latin square in which diets contained 40% B. napus or B. juncea CM or their air-classified fractions and 60% basal diet. Digesta data from pigs fed the N-free diet served to subtract basal endogenous AA losses. Eight ileal-cannulated barrows (32 kg initial BW) were fed the 8 diets at 2.7 times maintenance DE for eight 11-d periods. At the end of each period, feces were collected for 48 h, and ileal digesta were collected for two 12-h periods. The DE and calculated NE values and the apparent total tract digestibility (ATTD) of GE were 6.3%, 10.0%, and 7.8% greater (P < 0.001) for B. juncea CM than for B. napus CM; 6.1%, 10.8%, and 5.3% greater (P < 0.001) for the light-particle fraction than for parent CM; and 5.4%, 7.2%, and 3.8% lower (P < 0.001) for the heavy-particle fraction than for parent CM, respectively. The standardized ileal digestibilities (SID) of His, Ile, Val, Asp, and Tyr were greater (P < 0.05) for B. juncea CM than for B. napus CM. The SID of CP and AA were greater (P < 0.01) in the light-particle fraction than in the heavy-particle fraction. The SID of Trp, Glu, Pro, and Tyr were greater (P < 0.05) in the light-particle fraction than in parent CM. In conclusion, B. juncea CM had greater energy and AA digestibility than B. napus CM because of reduced fiber content. Air classification of CM increased its energy and AA digestibility in the light-particle fraction for pigs because of the reduced dietary fiber content and decreased particle size.

BACKGROUND: Tests sensitive to presymptomatic changes in Alzheimer's disease could be valuable for clinical trials. Accelerated long-term forgetting-during which memory impairment becomes apparent over longer periods than usually assessed, despite normal performance on standard cognitive testing-has been identified in other temporal lobe disorders. We assessed whether accelerated long-term forgetting is a feature of presymptomatic autosomal dominant (familial) Alzheimer's disease, and whether there is an association between accelerated long-term forgetting and early subjective memory changes.METHODS: This was a cross-sectional study at the Dementia Research Centre, University College London (London, UK). Participants were recruited from a cohort of autosomal dominant Alzheimer's disease families already involved in research at University College London, and had to have a parent known to be affected by an autosomal dominant Alzheimer's disease mutation, and not report any current symptoms of cognitive decline. Accelerated long-term forgetting of three tasks (list, story, and figure recall) was assessed by comparing 7-day recall with initial learning and 30-min recall. 7-day recognition was also assessed. Subjective memory was assessed using the Everyday Memory Questionnaire. The primary outcome measure for each task was the proportion of material retained at 30 min that was recalled 7 days later (ie, 7-day recall divided by 30-min recall). We used linear regression to compare accelerated long-term forgetting scores between mutation carriers and non-carriers (adjusting for age, IQ, and test set) and, for mutation carriers, to assess whether there was an association between accelerated long-term forgetting and estimated years to symptom onset (EYO). Spearman's correlation was used to examine the association between accelerated long-term forgetting and subjective memory scores.FINDINGS: Between Feb 17, 2015 and March 30, 2016, we recruited 35 people. 21 participants were mutation carriers (mean EYO 7·2 years, SD 4·5). Across the three tasks, we detected no differences between carriers and non-carriers for initial learning or 30-min recall. The proportion of material recalled at 7 days was lower in carriers than non-carriers for list (estimated difference in mean for list recall -30·94 percentage points, 95% CI -45·16 to -16·73; p=0·0002), story (-20·10, -33·28 to -6·91; p=0·0048), and figure (-15·41, -26·88 to -3·93; p=0·012) recall. Accelerated long-term forgetting was greater in carriers nearer to their estimated age at onset (p?0·01 for all three tests). Mutation carriers' 7-day recognition memory was also lower across all tasks (list [mean difference -5·80, 95% CI -9·96 to -2·47; p<0·01], story [-6·84, -10·94 to -3·37; p<0·01], and figure [-17·61, -27·68 to -7·72; p<0·01] recognition). Subjective memory scores were poorer in asymptomatic carriers compared with non-carriers (adjusted difference in means 7·88, 95% CI 1·36 to 14·41; p=0·016), and we found a correlation between accelerated long-term forgetting and subjective memory in mutation carriers.INTERPRETATION: Accelerated long-term forgetting is an early presymptomatic feature of autosomal dominant Alzheimer's disease, which appears to pre-date other amnestic deficits and might underpin subjective memory complaints in Alzheimer's disease. Accelerated long-term forgetting testing might be useful in presymptomatic Alzheimer's disease trials.FUNDING: MRC, NIHR, Alzheimer's Research UK, Dementias Platform UK, Dunhill Medical Trust, ERUK, Great Western Research, Health Foundation, Patrick Berthoud Trust.

We noticed that disease-related amyloidogenic proteins and especially cellular prion proteins have the highest proportion of incompletely desolvated backbone H bonds among soluble proteins. Such bonds are vulnerable to water attack and thus represent structural weaknesses. We have measured the adsorption of proteins onto phospholipid bilayers and found a strong correlation between the extent of underwrapping of backbone H bonds in the native structure of a protein and its extent of deposition on the bilayer: the less the H bond wrapping, the higher the propensity for protein-bilayer binding. These observations support the proposition that soluble proteins with amyloidogenic propensity and membrane proteins share a pervasive building motif: the underwrapped H bonds. Whereas in membrane proteins, this motif does not signal a structural vulnerability, in soluble proteins, it is responsible for their reactivity.

Acoustic Radiation Force Impulse (ARFI) imaging is a method for characterizing local variations in tissue mechanical properties. In this method, a single ultrasonic transducer array is used to both apply temporally short localized radiation forces within tissue and to track the resulting displacements through time. Images of tissue displacement immediately after force cessation, maximum tissue displacement, the time it takes for the tissue to reach its maximum displacement, and the recovery time constant of the tissue are generated from the ARFI data sets. The information in each of these images demonstrates good agreement with matched B-mode images. The study presented here was designed to evaluate the relationship between changes in these ARFI parameters with known tissue mechanical properties in vivo. Utilizing a modified Siemens Elegra scanner with a 75L40 transducer array, ARFI images of vastus medialis muscle were generated in three of the authors under four levels of activation (0, 5.7, 14.5, and 23.3 N-m). Four ARFI datasets were acquired for each loading condition. The observed trends were that displacement magnitude, the time it took for the tissue to reach its maximum displacement, and recovery time constant decreased with increasing load (i.e., increasing muscle stiffness). Significant differences were observed between load levels and subjects for all parameters (p<0.01). The results indicate that ARFI imaging may be capable of quantifying tissue stiffness in real-time measurements, although further investigation is required.

This study was an attempt to observe the effects of temperature on adsorption and one-step growth of the virus N-1 infecting the nitrogen-fixing cyanobacterium Nostoc muscorum. Adsorption rate was found to maximum at 40 degrees C whereas no adsorption occurred at 10 degrees C. The Q10 value was about 2.03 and the energy of activation, Ea was 16.3 kcal/mole for the adsorption process. The development cycle of the virus was temperature sensitive. With increase in temperature, a gradual increase in inhibition of virus yield i.e. 8.33% at 30 degrees C, 35.3% at 35 degrees C and complete inhibition at 40 degrees C was observed. Out of 7 h latent period, the early 4 h were temperature sensitive and heat treatment had a reversible inhibitory effect on virus development. The temperature treatment did not affect the rise period but burst-size was reduced.

Black mustard, Brassica nigra, is highly variable in both constitutive resistance and induction response following damage by herbivores. A focal population from Ithaca, New York, was used to test the following two predictions of optimal defense theory: (1) that allocation to resistance will reduce plant performance in the absence of herbivores; and (2) that induction response will be negatively correlated with constitutive resistance. The experiment consisted of a half-sib mating design with 47 paternal families and four dams per sire, fully crossed with a damage treatment consisting of 25% leaf removal by the cabbage white butterfly, Pieris rapae, when plants had four leaves. Leaf trichome density, sinigrin concentration, and glucobrassicin concentration were 38%, 19%, and 16% higher, respectively, for the seventh leaf of damaged plants. Paternal families did not vary significantly in their induction response. Narrow-sense heritabilities were h(S)(2) = 0.51, 0.76, and 0.50 for constitutive leaf trichome density, sinigrin concentration, and glucobrassicin concentration, respectively. Positive genetic correlations were found between glucobrassicin concentration and days to first flower, suggesting a genetic cost of resistance. Induction responses were negatively correlated with constitutive allocation for leaf trichome density and sinigrin concentration. The results were therefore consistent with optimal defense theory, offering modest evidence for both predictions.

White Leghorn chickens were selected for 36 generations for high (HAS) or low (LAS) antibody response to SRBC 5 d after an intravenous challenge. The aim of this study was to investigate possible changes in reproductive soundness resulting from that selection. Age and BW at onset of lay (first egg), along with weight of the first egg, were recorded on 45 hens from each line. Intensity of lay was measured as the number of ovulations within a 15-d period over 15 sequential intervals (total 225 d). Three cycles of fertility also were assessed, coinciding with early, middle, and late production stages. For fertility of males and females within a line to be independently evaluated, roosters and hens were mated by artificial insemination to an unrelated control line of White Plymouth Rocks. Twenty roosters from each antibody line were considered, as well as the 45 hens. Pooled semen from the control line was used for mating the hens from the antibody lines. Hens from the LAS line commenced lay at a younger age (11.67±3.53 d; P<0.001), lighter BW (-169.46±40.20 g; P<0.001), and with greater intensity (2.68±0.25%; P=0.001) than those from the HAS line. Any differences in intensity thereafter were trivial between lines (P=0.42), with intensity decreasing sharply toward the end of the 7-mo production period in both lines. Length of fertility differed between hens of the antibody lines during the first cycle (3.35±0.85 d; P=0.002) and between roosters during the first (3.58±1.06 d; P=0.02) and second (3.38±1.07 d; P=0.03) cycles, with chickens from the LAS line having the longer length of fertility in both sexes. A correlated response in reproductive soundness to divergent selection for antibody response was observed. This may in part be due to differences in resource allocations, with particular impact on duration of fertility.

BACKGROUND: Cocaine and methamphetamine (METH) are two commonly abused drugs that have behavioral-stimulant properties. These stimulant effects are partially mediated by the dopaminergic system. Recent evidence has suggested that the histamine H(3) receptor (H(3)R) may modulate the release of dopamine induced by METH. The aim of the present study was to examine the role of H(3)R in the behavioral-stimulant effects of cocaine and METH in mice and monkeys.METHODS: Nonhabituated, experimentally na?ve mice (n = 5-6) were pretreated with the H(3)R agonist imetit 30 min before METH or cocaine, and activity was measured for 90 min. The behavioral-stimulant effects of METH and cocaine were also studied in squirrel monkeys (n = 3) under a fixed-interval schedule of stimulus termination. Monkeys were pretreated with imetit 30 min before the peak behavioral-stimulant doses of METH or cocaine derived from individual subjects.RESULTS: Pretreatment with imetit did not affect basal activity in mice. Imetit significantly attenuated the behavioral-stimulant effects of METH, but not cocaine. In monkeys, no dose of imetit tested significantly altered the behavioral-stimulant effects of METH or cocaine.CONCLUSION: These results suggest a role of H(3)R in the behavioral-stimulant effects of METH, but not cocaine, in mice and no role in monkeys.

Epstein-Barr-virus (EBV)-transformed lymphoblastoid B-cell lines were generated from peripheral blood lymphocytes of 55 patients with systemic lupus erythematosus (SLE) and 44 healthy relatives. All donors have previously been extensively characterized with regard to clinical, serologic, and genetic parameters. Here, peripheral blood lymphocytes and lines were characterized for cell surface antigens. Furthermore, autoantibody production and proliferation rate of the cell lines were monitored. A significant difference between patients and relatives was the lower proliferation rate of EBV-transformed cell lines of the SLE patients. All SLE cell lines are available for interested researches and can be obtained from the European Cell Bank, Salisbury, UK.

The ectodomain of influenza A matrix protein 2 (M2e) is a candidate for a universal influenza A vaccine. We used recombinant Hepatitis B core antigen to produce virus-like particles presenting M2e (M2e-VLPs). We produced the VLPs with and without entrapped nucleic acids and compared their immunogenicity and protective efficacy. Immunization of BALB/c mice with M2e-VLPs containing nucleic acids induced a stronger, Th1-biased antibody response compared to particles lacking nucleic acids. The former also induced a stronger M2e-specific CD4(+) T cell response, as determined by ELISPOT. Mice vaccinated with alum-adjuvanted M2e-VLPs containing the nucleic acid-binding domain were better protected against influenza A virus challenge than mice vaccinated with similar particles lacking this domain, as deduced from the loss in body weight following challenge with X47 (H3N2) or PR/8 virus. Challenge of mice that had been immunized with M2e-VLPs with or without nucleic acids displayed significantly lower mortality, morbidity and lung virus titers than control-immunized groups. We conclude that nucleic acids present in M2e-VLPs correlate with improved immune protection.

Importance: Patients with cancer who die soon after starting chemotherapy incur costs of treatment without the benefits. Accurately predicting mortality risk before administering chemotherapy is important, but few patient data-driven tools exist.Objective: To create and validate a machine learning model that predicts mortality in a general oncology cohort starting new chemotherapy, using only data available before the first day of treatment.Design, Setting, and Participants: This retrospective cohort study of patients at a large academic cancer center from January 1, 2004, through December 31, 2014, determined date of death by linkage to Social Security data. The model was derived using data from 2004 through 2011, and performance was measured on nonoverlapping data from 2012 through 2014. The analysis was conducted from June 1 through August 1, 2017. Participants included 26 946 patients starting 51 774 new chemotherapy regimens.Main Outcomes and Measures: Thirty-day mortality from the first day of a new chemotherapy regimen. Secondary outcomes included model discrimination by predicted mortality risk decile among patients receiving palliative chemotherapy, and 180-day mortality from the first day of a new chemotherapy regimen.Results: Among the 26 946 patients included in the analysis, mean age was 58.7 years (95% CI, 58.5-58.9 years); 61.1% were female (95% CI, 60.4%-61.9%); and 86.9% were white (95% CI, 86.4%-87.4%). Thirty-day mortality from chemotherapy start was 2.1% (95% CI, 1.9%-2.4%). Among the 9114 patients in the validation set, the most common primary cancers were breast (21.1%; 95% CI, 20.2%-21.9%), colorectal (19.3%; 95% CI, 18.5%-20.2%), and lung (18.0%; 95% CI, 17.2%-18.8%). Model predictions were accurate for all patients (area under the curve [AUC], 0.940; 95% CI, 0.930-0.951). Predictions for patients starting palliative chemotherapy (46.6% of regimens; 95% CI, 45.8%-47.3%), for whom prognosis is particularly important, remained highly accurate (AUC, 0.924; 95% CI, 0.910-0.939). To illustrate model discrimination, patients were ranked initiating palliative chemotherapy by model-predicted mortality risk, and observed mortality was calculated by risk decile. Thirty-day mortality in the highest-risk decile was 22.6% (95% CI, 19.6%-25.6%); in the lowest-risk decile, no patients died. Predictions remained accurate across all primary cancers, stages, and chemotherapies, even for clinical trial regimens that first appeared in years after the model was trained (AUC, 0.942; 95% CI, 0.882-1.000). The same model also performed well for prediction of 180-day mortality (AUC for all patients, 0.870 [95% CI, 0.862-0.877]; highest- vs lowest-risk decile mortality, 74.8% [95% CI, 72.7%-77.0%] vs 0.2% [95% CI, 0.01%-0.4%]). Predictions were more accurate than estimates from randomized clinical trials of individual chemotherapies or the Surveillance, Epidemiology, and End Results data set.Conclusions and Relevance: A machine learning algorithm using electronic health record data accurately predicted short-term mortality among patients starting chemotherapy. Further research is necessary to determine the generalizability and feasibility of applying this algorithm in clinical settings.

Hind-limb motor function in adult female Lewis rats with experimental autoimmune encephalomyelitis (EAE) was investigated using an objective behavioral measurement of motor ability. Rats were pretrained to avoid falling from the external surface of a power-driven running wheel. The performance of EAE-group rats on the wheel was then compared with that of saline and adjuvant controls immediately prior to the onset of clinical signs of EAE, and within 3 days of apparent recovery from EAE. Results indicate no apparent hind-limb motor deficit in the absence of overt clinical signs of EAE, despite histological evidence of severe inflammatory lesions persisting in the central nervous system (CNS) at the time of the post-recovery test. The remarkably transient nature of motor impairment is discussed within the context of a continuing search for the underlying cause(s) of clinical signs of EAE.

BACKGROUND: Viral load (VL) scale-up efforts have largely focused on laboratory systems, with less attention on facility-level strengthening of staff who facilitate VL testing. To address this gap we implemented a quality improvement (QI) program at 13 health facilities in central and southern Malawi.METHODS: QI program tools focused on patient and provider VL knowledge and clarification of site-level roles and responsibilities, including the designation of a VL 'focal person' to oversee all VL activities. T-tests were used to compare differences in VL testing before (November 2016-April 2017) and after (May 2017-November 2017) the intervention.RESULTS: The mean number of VL tests performed significantly increased after implementation of the QI program. Overall there was a 164% increase in the mean number of routine VL tests performed per month (p<0.001). Increased VL testing was sustained during the 6 months of follow-up.CONCLUSIONS: A simple QI program focused on improving VL knowledge among patients and providers, and clarifying staff roles at a facility level increased VL testing over a 6-month period. Further investigation is needed on whether this program can be scaled in different settings across sub-Saharan Africa and on the duration of follow-up required for sustained improvements in VL testing.

We describe an unusual pathway of human immunodeficiency virus type 1 reverse transcriptase resistance during therapy with tenofovir-emtricitabine, characterized initially by the mutations K70E and M184V and later by K70G and M184V, with the two mutations coexisting on the same viral genome. Phenotypic resistance to lamivudine, emtricitabine, abacavir, didanosine, and tenofovir was observed, whereas susceptibility to zidovudine and stavudine was preserved.

OBJECTIVE: quality of life issues are central to maternal health and well-being. Within the context of a study examining postnatal quality of life, we set out to translate into Brazilian Portuguese the Mother-Generated Index and ensure its cross-cultural adaption for use in a Brazilian context.DESIGN: the Mother-Generated Index, a subjective quality of life tool, underwent a validated process of translation and cultural adaptation: synthesis of two independently translated versions, back-translation and review by an expert committee was followed by testing of the preliminary tool with 30 mothers.SETTINGS: community-based study in city in north-eastern Brazil.PARTICIPANTS: 30 postpartum mothers, interviewed approximately 30 days after birth.FINDINGS: while the mothers understood the concept of identifying and scoring quality of life aspects, many did not grasp the concept behind the use of 'spending points' to produce a relative ranking of these aspects. We resolved this by giving the mothers 'spending beans' instead; beans are a regional food staple. This use of a physical 'currency' solved the problem.DISCUSSION: this modified approach was ratified by the committee of experts and used with success on a further sample of 91 mothers. The whole process aimed to ensure semantic equivalence of the translated tool, and following this process we concluded that face validity of the Brazilian Portuguese Mother-Generated Index was good. While considerable resources are required to ensure a robust process of translation and adaptation, this is necessary if valid and reliable tools are to be produced.IMPLICATIONS: We conclude that the Mother-Generated Index is a valid tool to measure quality of life among Brazilian postpartum mothers, as it allows a simple and understandable way of assessing the various dimensions involved in their quality of life. Moreover, the Mother-Generated Index can provide healthcare professionals the opportunity to become aware of all significant aspects of a woman's life after childbirth.

During a 8-month period, Helicobacter pylori infection has been studied in 140 patients who presented upper gastrointestinal symptoms. Helicobacter pylori was searched in antral biopsy specimens using standard bacteriological methods (Gram-staining, urea-test, culture) and histological staining techniques (hematoxylin-eosin-saffron, undifferentiated Giemsa). The global prevalence of H. pylori infection was 59%. The prevalence rates did not seem to differ with age and sex but H. pylori infection was significantly more frequent in patients with an active duodenal ulceration (30 of 41) compared with those with a normal endoscopy (21 of 47) (p < 0.02).

In the 1980s, individual thyroid doses and uncertainties were estimated for members of a cohort of children identified in 1965 in Utah and Nevada who had potentially been exposed to fallout from the Nevada Test Site. That reconstruction represented the first comprehensive assessment of doses received by the cohort and was the first large effort to assess the uncertainty of dose on an individual person basis. The data on dose and thyroid disease prevalence during different periods were subsequently used in an analysis to determine risks of radiogenic thyroid disease. This cohort has received periodic medical follow-up to observe changes in disease frequency and to reassess the previously reported radiation-related risks, most recently after a Congressional mandate in 1998. In a recent effort to restore the databases and computer codes used to estimate doses in the 1980s, various deficiencies were found in the estimated doses due to improperly operating computer codes, corruption of secondary data files, and lack of quality control procedures. From 2001 through 2004, the dosimetry system was restored and corrected and all doses were recalculated. In addition, two parameter values were updated. While the mean of all doses has not changed significantly, many individual doses have changed by more than an order of magnitude.

The isolation of pancreatic islets from collagenase-digested Wistar rat pancreas was shown by the sedimentation method at a unit gravity using Percoll solution with a density of 1.041 g/ml. The density of digested exocrine tissues was in the range of 1.013-1.041 g/ml, while that of purely isolated islets was in the narrow range of 1.066-1.075 g/ml. More than a hundred islets were obtained freely from each rat pancreas without any gross contamination of digested exocrine tissues. A significant increase was observed in glucose-stimulated insulin release from islet isolated with Percoll in the same pattern as that without Percoll. In addition to the well preserved morphology and function of pancreatic islets isolated by Percoll, the simplicity of the technique strongly commends the usefullness of this method.

In this paper, we examine the potential of electromyographic (EMG) and acceleration measurements in discriminating patients with Parkinson's disease (PD) from healthy persons. Two types of muscle contractions are examined: static contractions of biceps brachii muscles and elbow extension movements. Twelve features are extracted from static and ten features from extension measurements. These features describe signal morphology and nonlinear characteristics, power spreading in EMG wavelet scalograms and spectral coherence. Principal component approach is applied separately for static and extension trial to reduce the number of features before discrimination. The discrimination between subjects is done in a two-dimensional space by applying cluster analysis to the best discriminating principal components. The discrimination power of the used method was estimated with EMG and acceleration data measured from 56 patients with PD and 59 healthy controls. In the cluster analysis, three clusters were formed: one cluster with most (85%) of the healthy persons and two clusters with 80% of patients. Patients were divided into two clusters based on their type of motor disability (problems during movement and/or static contraction). Discrimination results show that EMG and acceleration measurements are potential for discriminating patients with PD from healthy persons. Furthermore, they have potential in the objective clinical assessment of PD.

BACKGROUND: Pro-vitamin A-biofortified maize snacks with added leafy vegetable may have a potential as nutritious and health-promoting products, especially in addressing vitamin A deficiency, which is prevalent in developing regions. The objective of the study was to determine the effects of adding amaranth leaf powder on the physical, antioxidant properties and pro-vitamin A content of extruded pro-vitamin A-biofortified maize snacks. Extruded snacks were processed using four pro-vitamin A-biofortified maize varieties that were composited with amaranth leaf powder at 0%, 1% and 3% (w/w) substitution levels.RESULTS: At higher amaranth concentration, the expansion ratio of the snacks decreased, while their hardness increased by as much as 93%. The physical quality of the snacks may therefore need improvement. As amaranth was increased, the phenolic content and antioxidant activity of the snacks increased as well as the pro-vitamin A content.CONCLUSION: Pro-vitamin A-biofortified maize with added amaranth has a potential for use in nutritious and healthy extruded snacks. There are limited studies reporting on processing pro-vitamin A maize with complementary plant foods, which is common with white maize in southern Africa; thus the current study serves as a baseline.

Until recently, cave bears were believed to have only inhabited Europe. However, recent morphological evidence suggests that cave bears' geographic range extended as far east as Transbaikalia, Eastern Siberia. These Asian cave bears were morphologically distinct from European cave bears. However, how they related to European lineages remains unclear, stressing the need to assess the phylogenetic and phylogeographic relationship between Asian cave bears and their European relatives. In this work, we address this issue using a 227 base-pair fragment of the mitochondrial control region obtained from nine fossil bone samples from eight sites from the Urals, Caucasus, Altai Mountains, Ukraine and Yana River region in Eastern Siberia. Results of the phylogenetic analyses indicate that (i) the cave bear from the Yana River is most closely related to cave bears from the Caucasus region; (ii) the Caucasus/Yana group of bears is genetically very distinct from both European cave bears and brown bears, suggesting that these bears could represent an independent species; and (iii) the Western European cave bear lineage reached at least temporarily to the Altai Mountains, 7000 km east of their known centre of distribution. These results suggest that the diversity of cave bears was greater than previously believed, and that they could survive in a much wider range of ecological conditions than previously assumed. They also agree with recent studies on other extinct and extant species, such as wolves, hyenas and steppe bison, which have also revealed higher genetic and ecological diversity in Pleistocene populations than previously known.

We studied host factors that could predispose women to develop recurrent vulvovaginal candidiasis (RVVC), including glycemia, insulin resistance, chronic stress, antioxidant capacity, overall immune status, local inflammation and vaginal microbiota. The presence of yeasts in vaginal culture was screened in 277 women, with or without signs and symptoms of VVC and RVVC. The presence of an inflammatory process and microbiota were analyzed through vaginal bacterioscopy and cervical-vaginal cytology, respectively. Fasting-blood samples were collected by standard venipuncture for biochemical analyses. Flow cytometry was employed to obtain the T helper/T cytotoxic lymphocyte ratio, and insulin resistance was assessed by the HOMA index (HI). Yeasts were isolated from 71 (26%) women: 23 (32.4%) with a positive culture but without symptoms (COL), 22 (31%) in an acute episode (VVC), and 26 (36.6%) with RVVC. C. albicans was the main yeast isolated in all clinical profiles. The control group (negative culture) comprised 206 women. Diabetes mellitus and insulin resistance were more associated with the positive-culture groups (COL, VVC and RVVC) than with negative ones. The RVVC group showed lower mean levels of cortisol than the control group and lower antioxidant capacity than all other groups. The T Helper/T cytotoxic lymphocyte ratio was similar in all groups. The RVVC group showed a similar level of vaginal inflammation to the control group, and lower than in the COL and VVC groups. Only the CVV group showed a reduction in vaginal lactobacillus microbiota. Our data suggest that both chronic stress (decreased early-morning cortisol levels) and reduced antioxidant capacity can be host predisposing factors to RVVC.

The incidence, clinical manifestations, morbidity, and mortality of all adult hypernatremic patients hospitalized during a 6-month period were studied. The impact of age on this parameter was evaluated, and the velocity of correction that produced best clinical results was established. Records of all patients who were admitted or developed hypernatremia (Na+ > 150 mEq/L) were reviewed. Demographic characteristics of age, gender, associated diagnosis, length of stay, source of admission, treatment, and outcome were recorded. Of 3209 hospitalizations, 111 patients were hypernatremic (3.46%). Sixty-five were admitted with hypernatremia, and 45 developed hypernatremia while hospitalized. Fifteen had recurrent episodes of hypernatremia. Forty-nine had associated hypokalemia and six had hypercalcemia. The etiology was multifactorial and varied with age. Correction of the hypernatremia within 4 days produced significant improvement in mental status. The overall mortality was 48.6%, and age did not favorably influence mortality. Hypernatremia is a common disorder of elderly hospitalized patients, associated with high mortality and morbidity. Other electrolyte disorders that impair the kidney concentrating ability frequently are observed. The etiology is multiple, and febrile illness due to bacterial infections is the most common cause among the elderly. Prompt treatment of infections and increased water intake in this group of patients could prevent its development. Correction over a 72-hour period significantly improved recovery of mental functions.

The complex correlation structure of a collection of orthologous DNA sequences is uniquely captured by the "ancestral recombination graph" (ARG), a complete record of coalescence and recombination events in the history of the sample. However, existing methods for ARG inference are computationally intensive, highly approximate, or limited to small numbers of sequences, and, as a consequence, explicit ARG inference is rarely used in applied population genomics. Here, we introduce a new algorithm for ARG inference that is efficient enough to apply to dozens of complete mammalian genomes. The key idea of our approach is to sample an ARG of [Formula: see text] chromosomes conditional on an ARG of [Formula: see text] chromosomes, an operation we call "threading." Using techniques based on hidden Markov models, we can perform this threading operation exactly, up to the assumptions of the sequentially Markov coalescent and a discretization of time. An extension allows for threading of subtrees instead of individual sequences. Repeated application of these threading operations results in highly efficient Markov chain Monte Carlo samplers for ARGs. We have implemented these methods in a computer program called ARGweaver. Experiments with simulated data indicate that ARGweaver converges rapidly to the posterior distribution over ARGs and is effective in recovering various features of the ARG for dozens of sequences generated under realistic parameters for human populations. In applications of ARGweaver to 54 human genome sequences from Complete Genomics, we find clear signatures of natural selection, including regions of unusually ancient ancestry associated with balancing selection and reductions in allele age in sites under directional selection. The patterns we observe near protein-coding genes are consistent with a primary influence from background selection rather than hitchhiking, although we cannot rule out a contribution from recurrent selective sweeps.

BACKGROUND: The presence of osmotic gradients in the development of cerebral edema and the effectiveness of osmotherapy are well recognized. A modification of ventriculostomy catheters described in this article provides a method of osmotherapy that is not currently available. The reductive ventricular osmotherapy (RVOT) catheter removes free water from ventricular cerebrospinal fluid (CSF) by incorporating hollow fibers that remove water vapor, thereby providing osmotherapy without increasing osmotic load.OBJECTIVE: To increase osmolarity in the ventricular CSF through use of RVOT in vivo.METHODS: Twelve Yorkshire swine with contusional injury were randomized to external ventricular drainage (EVD) or RVOT for 12 hours. MR imaging was obtained. Serum, CSF, and brain ultrafiltrate were analyzed. Histology was compared using Fluor-Jade B and hematoxylin and eosin (H & E) stains.RESULTS: With RVOT, CSF osmolality increased from 292 ± 2.7 to 345 ± 8.0 mOsmol/kg (mean ± SE, P = 0.0006), and the apparent diffusion coefficient (ADC) in the injury region increased from 0.735 ± 0.047 to 1.135 ± .063 (P = 0.004) over 24 hours. With EVD controls, CSF osmolarity and ADC were not significantly changed. Histologically, all RVOT pigs showed no evidence of neuronal degeneration (Grade 1/4) compared to moderate degeneration (Grade 2.6 ± .4/4) seen in EVD treated animals (P = 0.02). The difference in intracranial pressure (ICP) by area under the curve approached significance at P = .065 by Mann Whitney test.CONCLUSION: RVOT can increase CSF osmolarity in vivo after experimental traumatic brain injury (TBI). In anticipated clinical use, only a slight increase in CSF osmolarity may be required to reduce cerebral edema.

PURPOSE: To report a case of local transmission of invasive lobular carcinoma from a donor to a recipient in a keratolimbal allograft after cessation of systemic immunosuppressive therapy.METHODS: This is a case report including the clinicopathologic findings. Sections of the donor breast tumor and recipient conjunctival lesions were stained with hematoxylin and eosin. Immunohistochemical studies were performed using pancytokeratin, CK7, CK20, CAM 5.2, CD138, TTF1, estrogen receptor, progesterone receptor, GATA-3, GCDFP-15, and mammaglobin. Polymerase chain reaction-based DNA profiling of tumor cells was performed.RESULTS: Histopathologic examination revealed an infiltrate of atypical cells with large hyperchromatic nuclei consistent with carcinoma. Immunohistochemical analysis showed pancytokeratin, CK7, CAM 5.2, GATA-3, and estrogen receptor positivity and progesterone receptor absence, consistent with the previously determined phenotype of the donor's breast carcinoma. Results of polymerase chain reaction analysis were also consistent with the donor's tumor. After reduced dosing of tacrolimus and mycophenolate mofetil, 2 limbal tumors occurred in the recipient. The immunosuppressive treatment had been stopped completely before the appearance of the third lesion. The recipient had no history of malignancy, and she had routine screenings for breast cancer.CONCLUSIONS: We report a case of donor-derived breast carcinoma in a keratolimbal allograft recipient. The grafted tissue harbored donor-derived tumor cells for more than 4 years after surgery even after systemic immunosuppression was discontinued. Although no similar reports of tumor transfer could be found in the literature, this case suggests the need for increased stringency in donor selection and heightened surveillance for such tumor transmission.

Due to a continuous process of dismantling within the university system, the number of freelance anthropologists has increased. In 2011 a group of experienced anthropologists founded the "Workgroup Freelance Osteoanthropologists (AFOA)". Its members developed a codex and guidelines in order to ensure a high quality for serious anthropological work. Furthermore workshops take place to discuss methods and current issues. Short reports given in this article present a small selection of projects conducted by some members of the AFOA.

OBJECTIVES: The purpose of this study was to determine whether a heart failure (HF) management strategy using continuous intracardiac pressure monitoring could decrease HF morbidity.BACKGROUND: Patients with HF may experience frequent decompensations that require hospitalization despite intensive treatment and follow-up.METHODS: The COMPASS-HF (Chronicle Offers Management to Patients with Advanced Signs and Symptoms of Heart Failure) study was a prospective, multicenter, randomized, single-blind, parallel-controlled trial of 274 New York Heart Association functional class III or IV HF patients who received an implantable continuous hemodynamic monitor. Patients were randomized to a Chronicle (Medtronic Inc., Minneapolis, Minnesota) (n = 134) or control (n = 140) group. All patients received optimal medical therapy, but the hemodynamic information from the monitor was used to guide patient management only in the Chronicle group. Primary end points included freedom from system-related complications, freedom from pressure-sensor failure, and reduction in the rate of HF-related events (hospitalizations and emergency or urgent care visits requiring intravenous therapy).RESULTS: The 2 safety end points were met with no pressure-sensor failures and system-related complications in only 8% of the 277 patients who underwent implantation (all but 4 complications were successfully resolved). The primary efficacy end point was not met because the Chronicle group had a nonsignificant 21% lower rate of all HF-related events compared with the control group (p = 0.33). A retrospective analysis of the time to first HF hospitalization showed a 36% reduction (p = 0.03) in the relative risk of a HF-related hospitalization in the Chronicle group.CONCLUSIONS: The implantable continuous hemodynamic monitor-guided care did not significantly reduce total HF-related events compared with optimal medical management. Additional trials will be necessary to establish the clinical benefit of implantable continuous hemodynamic monitor-guided care in patients with advanced HF.

A new generation of Atraumatic (blunt tipped) needles now exists in addition to the traditional Quincke (cutting tip) needles for LP. We wished to identify the optimal size and type of needle that should be used for this technique. Requirements include rapid and accurate transduction of CSF pressure and adequate flow rate for CSF collection purposes while minimizing headache and other neurologic sequelae. Different types of available Atraumatic and Quincke needles were tested in two models that simulated normal and raised CSF pressures. The flow rates and the time required for an accurate transduction of CSF pressure onto a standard spinal manometer through each needle type was measured. Overall, Atraumatic-tipped needles compared favorably with similarly sized Quincke needles tested. The 20-gauge needles displayed suitable flow and pressure transduction characteristics. Some of the 22-gauge Atraumatic needles rapidly measured CSF pressure, but their flow rates were only suitable for small volume CSF collection. There is extensive literature to support that neurologic sequelae are reduced using Atraumatic needles. Diagnostic LP can be easily and accurately performed by using a large Atraumatic needle with the potential for considerable reduction in post-LP headache and related neurologic sequelae.

OBJECTIVE: As of 2017, the pathobiology of gastric cancer (GC) is far from fully understood; consequently, new methods of basic and advanced research have been proposed and tested. The presence (GL1) vs absence (GL0) of malignant cells exfoliated in gastric lavage (GL) of GC patients was formerly evaluated with diagnostic intent but not for staging or prognostic assessment. We investigated this hitherto unreported application of cytopathology.METHODS: GL was preoperatively and prospectively collected from 80 GC patients and cytologically analysed. The results were compared with the classic clinicopathological features of GC and related to survival. The prognostic value of GL1 was assessed through univariate and multivariate analyses.RESULTS: GL1 was detected in 36 samples (45%) and correlated with advanced tumour depth (T3-T4), lymphatic metastasis (N+), distant metastasis (M1) and lymphovascular invasion (LVI1; P=.0317, .0024, .003 and .0028, respectively). Overall survival (OS) was significantly shorter for GL1 (23 months) vs GL0 patients (42 months; P=.005) and GL1 vs GL0 T1 subjects (12.6 vs 47.8 months, P=.0029). Univariate analysis revealed that GL1, N+, M1, LVI1 and advanced stage were significantly associated with OS. Multivariate analysis assessed GL1 as the only independent prognostic factor for worse OS and progression-free survival (P=.0013 and .0107).CONCLUSIONS: In the present study, GL1 was correlated with advanced disease, aggressive tumour behaviour and poor prognosis. Although additional studies are needed to confirm these findings, the GL0/GL1 classification can be applied to GC patients to achieve higher accuracy in staging, prognostic stratification and treatment selection.

Peripheral nerve injury due to trauma, disease, and certain toxins sometimes produces abnormal (neuropathic) pain syndromes that are chronic and refractory to standard analgesics. Knowledge of the mechanisms that produce neuropathic pain and the ability to search for new drugs to control it have been greatly advanced by the introduction of rat models of post-traumatic painful peripheral neuropathy. There are currently three models of neuropathic pain in the rat that are widely used. The procedures to create these models and the behavioral assays used to quantify the resulting neuropathic pain symptoms are described in this unit: the chronic constriction injury (CCI) model, the partial sciatic ligation (PSL) model, and the spinal nerve ligation (SNL) model. Four kinds of abnormal pain sensations are commonly measured to assess the outcome: heat-hyperalgesia, mechano-hyperalgesia, mechano-allodynia, and cold-allodynia.

Elevated superoxide formation in cardiac extracts of apolipoprotein E-knockout (apoE-KO) mice has been reported. In addition, we previously reported that hypoxia increased oxidative stress in the aortas of apoE-KO mice, although we did not examine the effect of hypoxia on the heart. The aim of this study was to investigate the effect of chronic hypoxia on the left ventricular (LV) remodeling in apoE-KO mice treated with or without an angiotensin II receptor blocker. Male apoE-KO mice (n=83) and wild-type mice (n=34) at 15 weeks of age were kept under hypoxic conditions (oxygen, 10.0+/-0.5%) and treated with olmesartan (3 mg/kg/day) or vehicle for 3 weeks. Although LV pressure was not changed, hypoxia caused hypertrophy of cardiomyocytes and increased interstitial fibrosis in the LV myocardium. Furthermore, nuclear factor-kappaB (NF-kappaB) and matrix metalloproteinase (MMP)-9 activities were increased in apoE-KO mice exposed to chronic hypoxia. Olmesartan effectively suppressed the 4-hydroxy-2-nonenal protein expression and NF-kappaB and MMP-9 activities, and preserved the fine structure of the LV myocardium without affecting the LV pressure. In conclusion, olmesartan reduced oxidative stress, and attenuated the hypoxia-induced LV remodeling, in part through the inhibition of NF-kappaB and MMP-9 activities, in apoE-KO mice.

BACKGROUND: Oxidative stress and inflammation can be altered by dietary factors in various species. However, little data are available in true carnivorous species such as domestic cats. As numerous anti-inflammatory and anti-oxidative additives become available and might be of use in cats with chronic low-grade inflammatory diseases, the current study aimed to develop a model of diet-induced inflammation by use of two opposite diets. It was hypothesized that a high fat diet enhanced in n-6 PUFA and with lower concentrations of antioxidants would evoke inflammation and oxidative stress in domestic cats.RESULTS: Sixteen healthy adult cats were allocated to two groups. One group received a moderate fat diet, containing pork lard and salmon oil (AA:(EPA + DHA) ratio 0.19) (MFn-3), while the other group was fed a high fat diet, containing pork lard and chicken fat (AA:(EPA + DHA) ratio 2.06) (HFn-6) for 12 weeks. Prior to and 2, 4, 6, 8, 10 and 12 weeks after starting the testing period, blood samples were collected. Erythrocytic fatty acid profile showed clear alterations in accordance to the dietary fatty acid profile. Serum thiobarbituric acid reactive substances was higher when fed MFn-3 compared to the HFn-6, suggesting augmented oxidative stress. This was associated with a reduced serum vitamin E status, as serum á-tocopherol concentrations were lower with MFn-3, even with higher dietary levels of vitamin E. Serum cytokine and serum amyloid A concentrations were not influenced by diet.CONCLUSION: These results point towards a resistance of cats to develop dietary fat-induced inflammation, but also suggest a high susceptibility to oxidative stress when fed a fish oil-supplemented diet even with moderate fat level and additional vitamin E.

There is little data regarding the diagnostic ability of neutrophil-lymphocyte ratio (NLR) in acute coronary syndrome. Therefore, we aimed to assess the predictive value of NLR in the discrimination of troponin-positive patients. We enrolled 244 patients (mean age 61.3 ± 11.7 years, 70.5% male) who presented to the emergency service with acute chest pain. Patients were divided into 2 groups based upon the troponin positivity in the 12- to 24-hour follow-up. The admission NLR was significantly higher in the troponin-positive group (5.49 ± 4.01 vs 2.40 ± 1.36, P < .001). A cutoff point of 2.80 for NLR measured on admission had 79% sensitivity and 73% specificity in predicting follow-up troponin positivity. After multivariate analysis, admission NLR and glucose levels remained significant predictors of follow-up troponin positivity. In conclusion, NLR may be an early diagnostic marker in emergency service for discriminating patients who will have a positive troponin test in the follow-up.

Paediatric patients with the syndrome of an inappropriate antidiuretic hormone secretion (SIADH), as a manifestation of inflammatory demyelinating disorders of the central nervous system, have been rarely described until now, in only a few cases of neuromyelitis optica spectrum disorders (NMOSDs). We present a case of relapsing SIADH associated with NMOSD, in an anti-aquaporin-4 antibody positive 14-year-old girl, who is, to our best knowledge, the first reported paediatric patient with relapsing SIADH and NMOSD. Additionally, our case further supports the notion that paediatric encephalomyelitis associated with SIADH should suggest the diagnosis of NMOSD.

The experiment reported here tests the Lexical Orthography Hypothesis, that is, the notion that the output of the lexical phonology is the most promising phonological depth for an exhaustive representation of tone by means of diacritics in the orthography of atone language. We conducted a controlled classroom experiment with 97 secondary school pupils learning written Kabiye, a Gur language of northern Togo. After testing their baseline skills in writing the standard orthography, the pupils participated in an eleven-hour transition course spread over three weeks in four parallel groups: DEEP (an experimental orthography representing the input of the lexical phonology), LEXICAL (representing the output of the lexical phonology), PHONEMIC (representing a level between the output of the lexical phonology and the output of the post-lexical phonology), and a control group. On the final day of the experiment, we tested their acquired skills in a dictation exercise. The results show that the LEXICAL group outperforms the other groups in three of the error types associated with adding diacritics, although they performed less well on the error type associated with writing long vowels. This initial evidence supporting the Lexical Orthography Hypothesis needs confirmation with reading and writing experiments on a variety of other tone languages.

Failure to maintain organelle genome stability has been linked to numerous phenotypes, including variegation and cytosolic male sterility (CMS) in plants, as well as cancer and neurodegenerative diseases in mammals. Here we describe a next-generation sequencing approach that precisely maps and characterizes organelle DNA rearrangements in a single genome-wide experiment. In addition to displaying global portraits of genomic instability, it surprisingly unveiled an abundance of short-range rearrangements in Arabidopsis thaliana and human organelles. Among these, short-range U-turn-like inversions reach 25% of total rearrangements in wild-type Arabidopsis plastids and 60% in human mitochondria. Furthermore, we show that replication stress correlates with the accumulation of this type of rearrangement, suggesting that U-turn-like rearrangements could be the outcome of a replication-dependent mechanism. We also show that U-turn-like rearrangements are mostly generated using microhomologies and are repressed in plastids by Whirly proteins WHY1 and WHY3. A synergistic interaction is also observed between the genes for the plastid DNA recombinase RECA1 and those encoding plastid Whirly proteins, and the triple mutant why1why3reca1 accumulates almost 60 times the WT levels of U-turn-like rearrangements. We thus propose that the process leading to U-turn-like rearrangements may constitute a RecA-independent mechanism to restart stalled forks. Our results reveal that short-range rearrangements, and especially U-turn-like rearrangements, are a major factor of genomic instability in organelles, and this raises the question of whether they could have been underestimated in diseases associated with mitochondrial dysfunction.

BACKGROUND: In a population-based multicenter case-control study of diet, life-style, and gastric cancer a large series of adults, aged 30-75 years (mean 58.9 years), were randomly sampled from the general population in 3 areas of Central-Northern Italy.AIM OF THE STUDY: To evaluate the relationship between plasma levels of antioxidant vitamins and cholesterol, and sociodemographic characteristics, life-style factors, and dietary intake of selected nutrients in a sample of the Italian population.METHODS: A fasting blood sample was available for 945 subjects (553 men, 392 women). The plasma concentrations of ascorbic acid, carotene, retinol, alpha-tocopherol, and cholesterol were determined by a centralized laboratory. All participants answered to a detailed questionnaire collecting information on sociodemographic and anthropometric characteristics, smoking, alcohol drinking, and dietary habits. Covariance analysis models, with post hoc Dunnett tests, including terms for age, sex, study center, and period of blood drawing, were used for selected multiple-way comparisons of mean values of plasma nutrients.RESULTS: Mean plasma values of retinol were higher among men while women had higher levels of plasma carotene, ascorbic acid, alpha-tocopherol, and cholesterol. Plasma carotene levels showed an inverse association with body mass index, alcohol consumption, and smoking and a positive association with social class. Carotene concentrations were higher in plasma samples obtained in spring/summer, while ascorbic acid levels were higher in autumn/winter. Partial correlation co-efficients between plasma vitamin levels showed a strong correlation between carotene and ascorbic acid (0.69 in men; 0.74 in women), between carotene and alpha-tocopherol (0.44; 0.37), and between alpha-tocopherol and ascorbic acid (0.45; 0.41). Plasma alpha-tocopherol and retinol correlated with plasma cholesterol. On the other hand, plasma carotene and ascorbic acid were correlated with their estimated dietary intakes, while the intakes of other nutrients, as expected, correlated rather poorly with the respective plasma concentrations.CONCLUSIONS: Socio-economic factors, life-style, and specific nutrient intake, in addition to gender, are related to nutrient plasma levels in Italian adults and may provide specific suggestions for the prevention of chronic diseases.

Two epidemiological studies were conducted from August 1997 to May 1998: a case-control study to identify herd level risk factors for antibodies to Aujeszky's disease virus (ADV) in sows in the state of Yucatan, Mexico and a cross-sectional study to determine the prevalence of antibodies against ADV in fattening pigs. In the case-control study, data on herd management and biosecurity were obtained from all the 27 ADV known field-virus-seropositive farms (cases) and 62 randomly selected seronegative farms (controls) by questionnaire. Breeding animals of these seropositive farms had received a gE-deletion vaccine. In the cross-sectional study, 26 farrow-to-finish farms of the 27 seropositive farms were used and blood samples taken from 60 fattening pigs per herd (15 pigs for each stage of production). Serum samples were analyzed by the screening-ELISA and gE-ELISA tests. In the case-control study, three of the 15 risk factors were significant. Odds ratios for distance to the nearest farm (< or = 2.5km), not sampling for the detection of ADV and herds with origin of breeding animals within the state were 9.5, 18.1 and 8.7. In the cross-sectional study, 11 (42.3%) of the 26 sampled farms were seropositive to vaccine antibodies. None of the piglets were positive to antibodies against field virus risk--suggesting that the strategy of vaccinating only the breeding animals reduced the ADV infection of the piglets.

An important property of Cox regression model is that the estimation of regression parameters using the partial likelihood procedure does not depend on its baseline survival function. We call such a procedure baseline-free. Using marginal likelihood, we show that an baseline-free procedure can be derived for a class of general transformation models under interval censoring framework. The baseline-free procedure results a simplified and stable computation algorithm for some complicated and important semiparametric models, such as frailty models and heteroscedastic hazard/rank regression models, where the estimation procedures so far available involve estimation of the infinite dimensional baseline function. A detailed computational algorithm using Markov Chain Monte Carlo stochastic approximation is presented. The proposed procedure is demonstrated through extensive simulation studies, showing the validity of asymptotic consistency and normality. We also illustrate the procedure with a real data set from a study of breast cancer. A heuristic argument showing that the score function is a mean zero martingale is provided.

OBJECTIVES: Tanezumab, a monoclonal antibody, inhibits nerve growth factor and reduces chronic pain. This randomised, double-blind, controlled multicentre study was conducted to evaluate the efficacy and safety of tanezumab added to oral diclofenac sustained release (DSR) in patients with hip or knee osteoarthritis (OA) pain.METHODS: Patients (N=604) with moderate to severe knee or hip OA tolerating stable DSR were randomised and treated with DSR 75 mg twice daily combined with intravenous tanezumab 10, 5 or 2.5 mg or placebo at weeks 0, 8 and 16. Co-primary efficacy endpoints (Western Ontario and McMaster Universities OA Index (WOMAC) Pain and Physical Function subscales and patient's global assessment of OA) were assessed at week 16.RESULTS: All co-primary endpoints were significantly improved for all tanezumab+DSR groups versus placebo+DSR (p?0.039). The incidence of adverse events of abnormal peripheral sensation was lower than in previous tanezumab trials. No new safety signals emerged. Overall incidence of adverse events was higher with tanezumab+DSR (45.2%-49.7%) than with placebo+DSR (34.9%); serious adverse event rates were similar across treatments (5.3%-7.6%). Osteonecrosis was reported in six of 452 patients with tanezumab+DSR (1.3%), but an external adjudication committee did not confirm osteonecrosis in any patient.CONCLUSIONS: Addition of tanezumab to DSR resulted in significant improvements in pain, function and global assessments in patients with OA. Although no new safety signals were observed, the higher incidence of adverse events in the tanezumab+diclofenac group suggests that combination therapy is unfavourable. Further investigations of tanezumab monotherapy for OA pain treatment are required.CLINICAL TRIAL REGISTRATION NUMBER: NCT00864097.

Carbon monoxide (CO), an activator of soluble guanylate cyclase (SGC) and generated enzymatically by heme oxygenases (HO), is considered to function as an intra- and intercellular neuromodulator or neurotransmitter in the central and peripheral nervous systems. HO-2 is the constitutive isoform of HO and is more prevalent in nervous tissues than in the other peripheral tissues. Because previous studies have demonstrated different distributions of HO-2 in the retina depending on the species of animals, the aim of this study was to identify which cell types of the monkey retina express HO-2. The expression of HO-2 protein was examined in monkey retina by Western blot analysis. Immunoblottings from monkey homogenates revealed a single clear protein band with a molecular mass of 36 kDa that is corresponding to rat HO-2. Immunoreactivity of HO-2 was found in the perikarya of ganglion cells. Density of immunoreactive ganglion cells was higher in the central area of retina than in the peripheral retina, and somata of larger ganglion cells were stained more densely than smaller ones. In electron microscopy, immunoreactivity of HO-2 was localized on the membrane of the endoplasmic reticulum and the nuclear outer membrane of the ganglion cells. By contrast, inner plexiform layer, inner nuclear layer and outer nuclear layer were devoid of HO-2 immunoreactivity. cGMP were strongly localized in all of ganglion cells. Some cells contributed to the relatively faint cGMP staining were seen in the inner nuclear layer. In combination of HO-2 and cGMP immunocytochemistry, the overlap of co-localization of HO-2 and cGMP would suggest that HO-2 in the ganglion cells would serve as a source for CO generation and CO could serve as a gaseous signaling molecule modulator of neural activity in the retina of monkey.

An 80,000 base-pair region from the gene locus encoding the variable regions of the human immunoglobulins of the kappa type (V kappa genes) was cloned and analysed. The region comprises five V kappa sequences of subgroup I and one interspersed V kappa pseudogene of subgroup II. The six genes and pseudogenes are arranged at different distances but in the same orientation. The organization of the cluster can be explained by a series of amplification steps; the existence of a V kappa II pseudogene in a V kappa I gene cluster may have been the result of a transposition event; a final duplication step led to a second closely related copy of the cluster. From sequence data for altogether 16,000 base-pairs it appears that gene conversion-like events and subsequent selection contribute to both homogeneity and diversity of the V kappa repertoire.

Perinatal undernutrition affects the hippocampus, a brain region crucial for learning and memory. However, far less is known about the changes of dendritic spine density and morphology related to hippocampal synaptic plasticity. As dendritic spines are dynamic structures essential for synaptic plasticity and serve as the primary post-synaptic location of the excitatory neurotransmission that underlies learning and memory, the aim of the present study was to investigate whether the perinatal undernutrition affected hippocampal synaptic plasticity accompanied by the change of dendritic spines in anesthetized rats. An input-output curve was first determined using the measurements of field excitatory postsynaptic potential (fEPSP) slope in response to a series of stimulation intensities. Long-term potentiation (LTP) induced by high-frequency stimulation was recorded in the Schaffer collateral-CA1 pathway. Post-tetanic potentiation derived from the fEPSP slope was also measured immediately after LTP induction. Quantitative data of dendritic spines from hippocampal CA1 pyramidal cells were obtained using Golgi staining. The results showed that 50% perinatal food restriction (FR50) impaired the magnitude of LTP of the fEPSP slope in the Schaffer collateral-CA1 pathway. Additionally, FR50 reduced overall spine densities in both basal dendrites and apical dendrites of hippocampal CA1 pyramidal cells. Moreover, FR50 reduced type densities of thin and mushroom spines in apical dendrites, whereas a reduction in the type of mushroom spines was only observed in the basal dendrites of hippocampal CA1 pyramidal cells. These findings suggested that perinatal undernutrition decreased excitatory synaptic input and further affected the processing of information in a given network by selectively reducing the number of special dendritic spines. Thus, these changes in the density and the types of dendritic spines in CA1 pyramidal neurons may partly explain the impaired hippocampal synaptic plasticity as well as learning and memory disturbances commonly observed during undernourished rats.

Transmission electron microscopy is the gold standard for examination of photoreceptor outer segment morphology and photoreceptor outer segment abnormalities in transgenic animal models of retinal disease. Small vertebrates such as zebrafish and Xenopus laevis tadpoles have been used to generate retinal disease models and to study outer segment processes such as protein trafficking, and their breeding capabilities facilitate experiments involving large numbers of animals and conditions. However, electron microscopy processing and analysis of these very small eyes can be challenging. Here we present a methodology that facilitates processing of X. laevis tadpole eyes for electron microscopy by introducing an intermediate cryosectioning step. This method reproducibly provides a well-oriented tissue block that can be sectioned with minimal effort by a non-expert, and also allows retroactive analysis of samples collected on slides for light microscopy.

The Gulf-Coast tick, Amblyomma maculatum, possesses an elaborate set of selenoproteins, which prevent the deleterious effects from oxidative stress that would otherwise occur during feeding. In the current work, we examined the role of selenoprotein K (SelK) and selenoprotein M (SelM) in feeding A. maculatum by bioinformatics, transcriptional gene expression, RNA interference and antioxidant assays. The transcriptional expression of SelK did not vary significantly in salivary glands or midguts throughout the bloodmeal. However, there was a 58-fold increase in transcript levels of SelM in tick midguts. Ticks injected with selK-dsRNA or selM-dsRNA did not reveal any observable differences in egg viability but oviposition was reduced. Surprisingly, salivary antioxidant activity was higher in selenoprotein knockouts compared with controls, which is probably the result of compensatory transcriptional expression of genes involved in combating reactive oxygen species. In fact, quantitative real-time PCR data suggest that the transcriptional expression of catalase increased in ticks injected with selM-double-stranded RNA. Additionally, the transcriptional expression of selN decreased ?90% in both SelK/SelM knockdowns. These data indicate that SelK and SelM are salivary antioxidants but are not essential for tick survival or reproduction and are compensated by other antioxidant systems.

Chronic disseminated intravascular coagulation (DIC) is a well-known complication of aortic aneurysm. A 63-year-old man with bleeding tendency and a large palpable abdominal aortic aneurysm (AAA) was diagnosed as having fibrinolysis dominant DIC by the excessive activation of both coagulation and fibrinolysis (plasmin -alpha2 plasmin inhibitor complex concentration is usually >4 microg/ml). Although several treatments were tried, DIC could not be controlled until the patient was given combined therapy of danaparoid (1,250 U/12 h, bolus IV) and tranexamic acid (0.5 g x 3/day, oral administration). This therapy may be beneficial when control for bleeding is required without restricting the ambulatory movement of patients by continuous drip.

Methicillin-resistant Staphylococcus aureus is a major problem in the world, causing hospital acquired infections and the infections/pathogenesis in community. Lysostaphin is a novel therapeutic molecule to kill the multidrug-resistant S. aureus. Mature lysostaphin is a single polypeptide (approximately 27 kDa) chain metalloprotease glycylglycine endopeptidase, capable of specifically hydrolyzing penta-glycine crosslinks present in the peptidoglycan of the S. aureus cell wall. The mature lysostaphin gene of Staphylococcus simulans has been cloned and overexpressed in the cytoplasm of E. coli with amino terminal hexa-histidine as a fusion partner under the transcriptional control of bacteriophage T7 phi 10 promoter/lac operator and ribosome binding site. The transformed E. coli BL21 (lambdaDE3) cells produced catalytically active soluble (His)6-lysostaphin fusion protein in the cytoplasm representing approximately 20% of the total cellular proteins. The fusion protein was purified to homogeneity using a single chromatographic step of IMAC on Ni-NTA agarose. The present cloning, expression, and purification procedure of recombinant lysostaphin from a non-pathogenic organism E. coli enables preparation of large quantity of r-lysostaphin for structure function studies and evaluation of its clinical potential in therapy and prophylaxis of staphylococcal infections.

The contribution of phospholipase D (PLD) to the production of phosphatidic acid (PA) and diglyceride (DG) by C5a-stimulated human neutrophils has been studied. Membrane-associated 1-O-alkyl-phosphatidylcholine (alkyl-PC) was double labeled with 3H and 32P by incubating neutrophils with [3H]alkyl-lysoPC and alkyl-[32P]lysoPC. Upon stimulation with recombinant C5a, these labeled neutrophils produce 1-O-alkyl-phosphatidic acid (alkyl-PA) and, in the presence of ethanol, 1-O-alkyl-phosphatidyl-ethanol (alkyl-PEt), containing both 3H and 32P. Formation of radiolabeled alkyl-PEt parallels that of radiolabeled alkyl-PA and requires both extracellular Ca2+ and cytochalasin B. Furthermore, the 3H/32P ratios of alkyl-PA and alkyl-PEt formed during stimulation are very similar to that of th substrate alkyl-PC. These results demonstrate that, in C5a-stimulated neutrophils, alkyl-PA and alkyl-PEt are formed from alkyl-PC almost exclusively by PLD-catalyzed hydrolysis and transphosphatidylation, respectively. Upon C5a stimulation, neutrophils labeled with 3H and 32P also produce 1-O-[3H]alkyl-diglyceride [( 3H]alkyl-DG) and [32P]orthophosphate [( 32P]PO4), but not [32P]phosphocholine. [3H]Alkyl-DG and [32P]PO4 are formed in parallel, although temporally lagging behind alkyl-PA. Propranolol, a PA phosphohydrolase (PPH) inhibitor, decreases the formation of both [3H]alkyl-DG and [32P]PO4, although increasing alkyl-PA accumulation. These data support the conclusion that alkyl-DG is formed from alkyl-PC by the combined activities of PLD and PPH and not by phospholipase C (PLC). Furthermore, by using [3H]acyl-PC-labeled neutrophils, it is demonstrated that, like alkyl-PC, 1-acyl-PC is also degraded sequentially by PLD and PPH to 1-acyl-DG. Propranolol does not inhibit phosphoinositide-specific PLC and yet it causes almost complete inhibition of the total DG mass accumulation in C5a-stimulated neutrophils. We conclude that, in cytochalasin B-treated neutrophils stimulated with C5a, PLD-catalyzed hydrolysis of PC determines the levels of both PA and DG with potentially important ramifications for neutrophil-mediated defense functions.

BACKGROUND: Monoclonal immunoglobulins (paraproteins) are produced by B lymphocytes in lymphoproliferative disorders. A single monoclonal immunoglobulin is homogeneous in terms of its structure, and it can occur in human serum at high concentration and cause significant interference in laboratory assays.CASE: We present a case of an 84-year-old man who was admitted to the hospital for progression of dyspnea. Basic laboratory tests showed a serum concentration of conjugated bilirubin, measured using the diazo spectrophotometric method, which was much higher than that of total bilirubin. The cause of the discrepancy was attributed to analytical interference by monoclonal immunoglobulins, which helped establish a diagnosis of lymphoproliferative disorder.CONCLUSION: Monoclonal immunoglobulins are relatively rare in serum but are an important cause of analytical interference. Monoclonal immunoglobulins should always be considered a source of interference when unexpectedly high, low, or contradictory data are encountered, and appropriate confirmatory tests (electrophoresis, imunofixation) should be performed in such circumstances. Failure to do so can result in errors in diagnosis and inadequate treatment. Conversely, when samples contain abnormal and especially high monoclonal immunoglobulin levels, the biochemical data should be carefully examined for any discrepancies, such as paraprotein interference, and the results should be taken into consideration in patient management.Key words: paraproteins - lymphoproliferative disorders - bilirubin - interferenceThe authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 22. 1. 2016Accepted: 1. 12. 2016.

Effects of climate change on productivity of agricultural crops in relation to diseases that attack them are difficult to predict because they are complex and nonlinear. To investigate these crop-disease-climate interactions, UKCIP02 scenarios predicting UK temperature and rainfall under high- and low-CO(2) emission scenarios for the 2020s and 2050s were combined with a crop-simulation model predicting yield of fungicide-treated winter oilseed rape and with a weather-based regression model predicting severity of phoma stem canker epidemics. The combination of climate scenarios and crop model predicted that climate change will increase yield of fungicide-treated oilseed rape crops in Scotland by up to 0.5 t ha(-1) (15%). In contrast, in southern England the combination of climate scenarios, crop, disease and yield loss models predicted that climate change will increase yield losses from phoma stem canker epidemics to up to 50 per cent (1.5 t ha(-1)) and greatly decrease yield of untreated winter oilseed rape. The size of losses is predicted to be greater for winter oilseed rape cultivars that are susceptible than for those that are resistant to the phoma stem canker pathogen Leptosphaeria maculans. Such predictions illustrate the unexpected, contrasting impacts of aspects of climate change on crop-disease interactions in agricultural systems in different regions.

The gastrointestinal lumen is directly exposed to dietary contaminants, including patulin, a mycotoxin produced by moulds. Patulin is known to increase permeability across intestinal Caco-2 monolayers. This study aimed to determine the effect of patulin on permeability, ion transport and morphology in isolated rat colonic mucosae. Mucosal sheets were mounted in Ussing chambers and voltage clamped. Apical addition of patulin (100-500 ìM) rapidly reduced transepithelial electrical resistance (TEER) and increased permeability to [(14)C] mannitol (2.9-fold). Patulin also inhibited carbachol-induced electrogenic chloride secretion and histological evidence of mucosal damage was observed. To examine potential mechanisms of action of patulin on colonic epithelial cells, high-content analysis of Caco-2 cells was performed and this novel, quantitative fluorescence-based approach confirmed its cytotoxic effects. With regard to time course, the cytotoxicity determined by high content analysis took longer than the almost immediate reduction of electrical resistance in isolated mucosal sheets. These data indicate patulin is not only cytotoxic to enterocytes but also has the capacity to directly alter permeability and ion transport in intact intestinal mucosae. These data corroborate and extend findings in intestinal cell culture monolayers, and further suggest that safety limits on consumption of patulin may be warranted.

On August 25, 1974 the Taft-Hartley Amendments (Public Law 93-360) took effect and mandated that voluntary not-for-profit hospitals enter into the process of collective bargaining. The belated entrance of these hospitals into the process stems from a change in the Taft-Hartley Act of 1947 where the law specifically exempted voluntary hospitals from the National Labor Relations Act (NLRA) of 1935. In deleting this exemption, Congress extended labor relations protection to employees of voluntary hospitals and all other healthcare institutions except those under government and public ownership. As far as most hospital chief executive officers were concerned, labor relations would never be the same again.

The rationale of anti-androgen treatment for hirsutism is explained. The results are presented of treatment with cyproterone acetate of idiopathic hirsutism or hirsutism associated with polycystic ovarian disease. Ten out of 14 patients showed a good response after six months treatment. The response was dramatic in two patients.

Efficacy of the ureteric stricture (UC) treatment, using energy of the Holmium (in 55% patients) and the diode (in 45%) lasers was analyzed. In 37 (77%) patient spositive results were achieved already after one session, in 4 (8.3%) — one more session was performed in terms up to 3 mo after the first one, in 3 (6.3%) — two sessions were per' formed. Positive results were achieved in total in 91.6% patients. Application of laser energy in treatment of UC is effective and not dependent from the energy kind and con' stitutes a perspective method of the patient's endoscopic treatment.

Nacre achieves excellent mechanical properties with a relatively simple hierarchical structure. Analyses suggest that a significant gain in toughness is realized with a modest reduction in strength, with increasing levels of hierarchy. This study probes the role of different hierarchical length scales in governing the strength and modulus of nacre using a combination of bulk compression tests, microindentation and nanoindentation tests. The variability in the measured properties is assessed through Weibull analyses. The transition from elastic deformation is characterized using spherical indentation tests at the micro and nano scales together with a Herztian analysis. The modulus of the organic phase at different scales was deduced using indentation data and appropriate micromechanical models. The results show a minimal influence of length scales on elastic-plastic transitions, suggesting that initiation of plasticity occurs through a common biomineral sliding mechanism across length scales. However the ultimate strengths follow the trends of models for hierarchical materials, with the strength reducing by a factor of ~2 with each increase in level of hierarchy. The modulus of the organic phase was higher at the lowest scale, in contrast to an earlier study, indicating that confinement significantly modifies the effective properties of the organic.

G-protein-coupled receptor kinase 2 (GRK2) is a central regulator of G-protein-coupled receptor signaling. We report that Mdm2, an E3-ubiquitin ligase involved in the control of cell growth and apoptosis, plays a key role in GRK2 degradation. Mdm2 and GRK2 association is enhanced by beta(2)-adrenergic receptor stimulation and beta-arrestin. Increased Mdm2 expression accelerates GRK2 proteolysis and promotes kinase ubiquitination at defined residues, whereas GRK2 turnover is markedly impaired in Mdm2-deficient cells. Moreover, we find that activation of the PI3K/Akt pathway by insulin-like growth factor-1 alters Mdm2-mediated GRK2 degradation, leading to enhanced GRK2 stability and increased kinase levels. These data put forward a novel mechanism for controlling GRK2 expression in physiological and pathological conditions.

BACKGROUND: Most traditional Chinese herbal formulas consist of at least four herbs. Four-Agents-Decoction (Si Wu Tang) is a documented eight hundred year old formula containing four herbs and has been widely used to relieve menstrual discomfort in Taiwan. However, no specific effect had been systematically evaluated. We applied Western methodology to assess its effectiveness and safety for primary dysmenorrhoea and to evaluate the compliance and feasibility for a future trial.METHODOLOGY/PRINCIPAL FINDINGS: A randomised, double-blind, placebo-controlled, pilot clinical trial was conducted in an ad hoc clinic setting at a teaching hospital in Taipei, Taiwan. Seventy-eight primary dysmenorrheic young women were enrolled after 326 women with self-reported menstrual discomfort in the Taipei metropolitan area of Taiwan were screened by a questionnaire and subsequently diagnosed by two gynaecologists concurrently with pelvic ultrasonography. A dosage of 15 odorless capsules daily for five days starting from the onset of bleeding or pain was administered. Participants were followed with two to four cycles for an initial washout interval, one to two baseline cycles, three to four treatment cycles, and three follow-up cycles. Study outcome was pain intensity measured by using unmarked horizontal visual analog pain scale in an online daily diary submitted directly by the participants for 5 days starting from the onset of bleeding or pain of each menstrual cycle. Overall-pain was the average pain intensity among days in pain and peak-pain was the maximal single-day pain intensity. At the end of treatment, both the overall-pain and peak-pain decreased in the Four-Agents-Decoction (Si Wu Tang) group and increased in the placebo group; however, the differences between the two groups were not statistically significant. The trends persisted to follow-up phase. Statistically significant differences in both peak-pain and overall-pain appeared in the first follow-up cycle, at which the reduced peak-pain in the Four-Agents-Decoction (Si Wu Tang) group did not differ significantly by treatment length. However, the reduced peak-pain did differ profoundly among women treated for four menstrual cycles (2.69 (2.06) cm, mean (standard deviation), for the 20 women with Four-Agents-Decoction and 4.68 (3.16) for the 22 women with placebo, p = .020.) There was no difference in adverse symptoms between the Four-Agents-Decoction (Si Wu Tang) and placebo groups.CONCLUSION/SIGNIFICANCE: Four-Agents-Decoction (Si Wu Tang) therapy in this pilot post-market clinical trial, while meeting the standards of conventional medicine, showed no statistically significant difference in reducing menstrual pain intensity of primary dysmenorrhoea at the end of treatment. Its use, with our dosage regimen and treatment length, was not associated with adverse reactions. The finding of statistically significant pain-reducing effect in the first follow-up cycle was unexpected and warrants further study. A larger similar trial among primary dysmenorrheic young women with longer treatment phase and multiple batched study products can determine the definitive efficacy of this historically documented formula.TRIAL REGISTRATION: Controlled-Trials.com ISRCTN23374750.

Considering the poor medical conditions in some regions of China, this paper attempts to develop a simple and easy way to extract and process the bone features of blurry medical images and improve the diagnosis accuracy of osteoporosis as much as possible. After reviewing the previous studies on osteoporosis, especially those focusing on texture analysis, a convexity optimization model was proposed based on intra-class dispersion, which combines texture features and shape features. Experimental results show that the proposed model boasts a larger application scope than Lasso, a popular feature selection method that only supports generalized linear models. The research findings ensure the accuracy of osteoporosis diagnosis and enjoy good potentials for clinical application.

The objective of this study was to compare the concentrations of inflammatory cytokines in uterine flush and serum from healthy postpartum dairy cows and cows with clinical or subclinical endometritis. Clinical endometritis was diagnosed by observation of vaginal discharges (>50% pus) and subclinical endometritis was diagnosed by evaluation of uterine cytology (neutrophils >18%) at 4 weeks postpartum. Uterine flush was obtained from 48 cows at 4, 6, and 8 weeks postpartum for evaluation of tumor necrosis factor-á (TNF-á), interleukin (IL)-1â, IL-6, IL-8, and IL-10 concentrations. Serum samples were obtained from 34 cows just after calving and at 1, 2, 4, 6, and 8 weeks postpartum for evaluation of TNF-á, IL-1â, and IL-6 concentrations. Concentrations of TNF-á, IL-6, and IL-10 were greater (P < 0.05) in cows with clinical endometritis than in cows with subclinical endometritis and healthy controls, whereas concentrations of IL-8 in both cows with clinical and subclinical endometritis were greater (P < 0.005) than in controls. Overall, IL-6 and IL-10 concentrations decreased during the postpartum period. IL-1â concentrations in cows with clinical endometritis decreased (P < 0.0005) during the postpartum, whereas concentrations in cows with subclinical endometritis and controls did not change significantly with time; at 4 weeks postpartum, concentrations were greater (P < 0.0001) in cows with clinical endometritis. There were no significant effects of group, sampling time, or interaction on serum cytokine concentrations. In conclusion, cows with endometritis have greater inflammatory cytokine concentrations in uterine flush than healthy cows, but no differences were observed in serum.

BACKGROUND: Dietary salt is related to blood pressure (BP), and cardiovascular disease and increased sodium intakes have been shown to impair vascular function. The effect of salt on endothelial function postprandially is unknown.OBJECTIVE: The aim was to investigate the postprandial effect of dietary salt on endothelial function as measured by flow-mediated dilatation (FMD) and peripheral arterial tonometry in healthy subjects.DESIGN: Sixteen healthy, normotensive subjects received a meal with added salt (HSM; 65 mmol Na) and a control low-salt meal (LSM; 5 mmol Na) on 2 separate occasions in a randomized order. Endothelial function was measured while fasting and postprandially at 30, 60, 90, and 120 min by using FMD and reactive hyperemia peripheral arterial tonometry. BP was also measured.RESULTS: Baseline FMD, reactive hyperemia index (RHI), and BP values were similar across interventions. Overall FMD was reduced 2 h postprandially. FMD was significantly more impaired after the HSM than after the LSM at 30 min [HSM (mean ± SD): 3.39 ± 2.44%; LSM: 6.05 ± 3.21%; P < 0.01] and at 60 min (HSM: 2.20 ± 2.77%; LSM: 4.64 ± 2.48%; P < 0.01). No significant differences in BP or RHI were observed between meals.CONCLUSIONS: An HSM, which reflects the typical amount of salt consumed in a commonly eaten meal, can significantly suppress brachial artery FMD within 30 min. These results suggest that high salt intakes have acute adverse effects on vascular dilatation in the postprandial state. This trial was registered at www.anzctr.org.au/trial_view.aspx?ID=335115 as ACTRN12610000124033.

AIM: To examine the internal and external responsiveness of the Pressure Ulcer Scale for Healing (PUSH) tool for assessing the healing progress in acute and chronic wounds.BACKGROUND: It is important to establish the responsiveness of instruments used in conducting wound care assessments to ensure that they are able to capture changes in wound healing accurately over time.DESIGN: Prospective longitudinal observational study.METHOD: The key study instrument was the PUSH tool. Internal responsiveness was assessed using paired t-testing and effect size statistics. External responsiveness was assessed using multiple linear regression. All new patients with at least one eligible acute or chronic wound, enrolled in the Nurse and Allied Health Clinic-Wound Care programme between 1 December 2012 - 31 March 2013 were included for analysis (N = 541).RESULTS: Overall, the PUSH tool was able to detect statistically significant changes in wound healing between baseline and discharge. The effect size statistics were large. The internal responsiveness of the PUSH tool was confirmed in patients with a variety of different wound types including venous ulcers, pressure ulcers, neuropathic ulcers, burns and scalds, skin tears, surgical wounds and traumatic wounds. After controlling for age, gender and wound type, subjects in the 'wound improved but not healed' group had a smaller change in PUSH scores than those in the 'wound healed' group. Subjects in the 'wound static or worsened' group had the smallest change in PUSH scores. The external responsiveness was confirmed.CONCLUSION: The internal and external responsiveness of the PUSH tool confirmed that it can be used to track the healing progress of both acute and chronic wounds.

Insurance medicine is becoming more and more important. Currently, there are few countries in Europe where insurance medicine is recognised as an independent discipline; the Netherlands is one example. Since 2007 the "Specialist in Insurance Medicine and Medico-legal Expertise" is recognised in Belgium. This article will give an overview of the residency of Flemish physicians. By enactment, this consists of a theoretical and a practical section. This way of education should open broad possibilities in private and social insurance medicine, but also in the research sector.

OBJECTIVES: Soft tissue wounds with exposed bone often require extended healing times and can be associated with severe complications. We describe the ability of artificial dermis with autogenic adipose-derived stem cells (ADSCs) to promote the healing of wounds with exposed bone in a rat model.METHODS: Adipose tissues harvested from the bilateral inguinal regions of Wistar rats were used as ADSCs. Rats were randomly divided into control and ADSC groups to investigate the efficacy of ADSC transplantation for wound healing (n = 20 per group). Soft tissue defects were created on the heads of the rats and were covered with artificial dermis with or without the seeded ADSCs. Specimens from these rats were evaluated using digital image analysis, histology, immunohistochemistry, cell labeling, and real-time reverse-transcription polymerase chain reaction (real-time RT-PCR).RESULTS: The average global wound area was significantly smaller in the ADSC group than in the control group on days 3, 7, and 14 after surgery (p<0.05). After 14 days, the blood vessel density in the wound increased by 1.6-fold in the ADSC group compared with that in the control group (p<0.01). Real-time RT-PCR results showed higher Fgfb and Vegf expression levels at all time points, and higher Tgfb1 and Tgfb3 expression levels until 14 days after surgery in the ADSC group than in the control group (p<0.05).CONCLUSIONS: In wounds with exposed bone, autogenic ADSCs can promote vascularization and wound healing. Use of this cell source has multiple benefits, including convenient clinical application and lack of ethical concerns.

Alpha(IIb)beta(3) and alpha(v)beta(3) belong to the beta(3) integrin subfamily. Although the beta(3) subunit is a key regulator for the biosynthesis of beta(3) integrins, it remains obscure whether missense mutations in beta(3) may induce the same defects in both alpha(IIb)beta(3) and alpha(v)beta(3). In this study, it is revealed that thrombasthenic platelets with a His280Pro mutation in beta(3), which is prevalent in Japanese patients with Glanzmann thrombasthenia, did contain significant amounts of alpha(v)beta(3) (about 50% of control) using sensitive enzyme-linked immunosorbent assay. Expression studies showed that the His280Probeta(3) mutation impaired alpha(IIb)beta(3) expression but not alpha(v)beta(3) expression in 293 cells. To extend these findings, the effects of several beta(3) missense mutations leading to an impaired alpha(IIb)beta(3) expression on alpha(v)beta(3) function as well as expression was examined: Leu117Trp, Ser162Leu, Arg216Gln, Cys374Tyr, and a newly created Arg216Gln/Leu292Ser mutation. Leu117Trp and Cys374Tyr beta(3) mutations did impair alpha(v)beta(3) expression, while Ser162Leu, Arg216Gln, and Arg216Gln/Leu292Ser mutations did not. With regard to ligand binding function, Ser162Leu mutation induced especially distinct effects between 2 beta(3) integrins: it markedly impaired ligand binding to alpha(IIb)beta(3) but not to alpha(v)beta(3) at all. These data clearly demonstrate that the biosynthesis and the ligand binding function of alpha(IIb)beta(3) and those of alpha(v)beta(3) are regulated in part by different mechanisms. Present data would be a clue to elucidate the regulatory mechanism of expression and function of beta(3) integrins.

BACKGROUND: Previous studies have shown that the chronic open-angle glaucomas form a heterogeneous spectrum of diseases which have in common an open anterior chamber angle and glaucomatous optic nerve damage. Purpose of this study was to evaluate whether the appearance of the optic disc differs among the various types of primary open-angle glaucoma.METHODS: Color stereo optic disc photographs of 683 patients with primary open-angle glaucoma (POAG), and 481 normal eyes were morphometrically evaluated.RESULTS: Morphologic characteristics of the glaucoma types were as follows: Highly myopic POAG: secondary macrodiscs with abnormal shape; shallow, flat, concentric disc cupping; low frequency of disc hemorrhages; large parapapillary atrophy or myopic crescent; medium to low intraocular pressure. Juvenile-onset POAG: Optic disc of normal size and shape; deep and steep disc cupping; low frequency of broad rim notches or large disc hemorrhages; small parapapillary atrophy; high minimal and maximal intraocular pressure measurements. Age-related atrophic POAG: Optic disc of normal size and shape; shallow, flat and concentric disc cupping; medium to low frequency of disc hemorrhages; large parapapillary atrophy; medium to low intraocular pressure. Eyes with normal-pressure glaucoma: Optic disc of normal size and shape; deep and steep cupping; relatively small parapapillary atrophy; high frequency of disc hemorrhages and rim notches.CONCLUSIONS: These characteristics in the appearance of the optic disc may be helpful for clinical diagnosis and therapy and may give pathogenetic hints.

Analysis of 146,521 autopsy protocols in Moscow in 2001-2005 showed that the incidence pulmonary thromboembolism (PTE) as a direct cause of death was 3.8-7%. PTE was not diagnosed during life in 20-28% of the patients who had died at hospitals and in 68-72% of those who had died at home. PTE was 2-3 times more common in females than in males, the age of the deceased patients was old and senile. The sources of PTE were thrombi of the leg deep veins (93.2%), small pelvic veins (3.4%), and right heart chambers (1.1%). Thrombophilic states were one of the main causes of the poor course and outcome of PTE. Based on the studies performed, an algorithm was developed for the diagnosis and treatment, which was successfully used at the N.N. Burdenko Main Military Clinical Hospital. The introduction of the results could reduce the incidence venous thrombosis from 39% to 17%, that of cava-filter thromboses from 24% to 3.7%, and recurrent TELA from 15% to 5.6%.

PURPOSE: Technical quality assurance in diagnostic ultrasound is based on nationally standardized protocols which are mandatory for outpatient care in Germany. Quality assurance includes technical requirements for sonographic instruments based on limiting values for technical parameters and requirements for image quality and documentation. Certification by regional Associations of Statutory Health Insurance Physicians (Kassen?rztliche Vereinigungen - KV) requires acceptance tests to prove compliance with all required standards.MATERIALS AND METHODS: Between April 2009 und December 2011a total of 7457 acceptance tests were performed by the KV Westfalen-Lippe, which represents nearly 13 000 physicians in northwestern parts of Germany. Results of these acceptance tests are evaluated with respect to display mode or Doppler technique, anatomic region and year of manufacture of the instruments.RESULTS: In 2469 acceptance tests (33 %), objections were raised, mainly due to noncompliance with technical parameters. The frequency of objections strongly increases with the age of sonographic instruments and exceeds 90 % for instruments manufactured before 1995. In acceptance tests where compliance with technical parameters has been approved, insufficient documentation was reported in 6 % and inadequate image quality in 8 % of the tests. Frequency and cause of objections depend on display mode and anatomic region.CONCLUSION: The study shows that sonographic instruments of higher age often do not meet the required standards, and it can be expected that updates in technical equipment will result in improvements in diagnostic performance and clinical outcome.

BACKGROUND: Cystoid macular edema (CME) in AIDS patients with inactive cytomegalovirus (CMV) retinitis is an uncommon but potentially sight-threatening complication. The pathogenesis of CME in these patients is unclear. This study tries to identify possible risk factors by analyzing the charts of five patients.METHODS: Ten eyes of 5 patients that finally developed CME were followed for an average of 18 months. The initial retinal lesions, their response to antiviral treatment, the development of CME, and the patients' immune status were prospectively monitored.RESULTS: CMV retinitis was diagnosed at a median CD4+ count of 3 cells/mm3 (range 0-11). All eyes responded to the initial systemic anti-viral treatment. At the onset of CME, CMV retinitis was controlled by antiviral maintenance therapy in all patients [ganciclovir (n = 2), cidofovir (n = 2), foscarnet (n = 1)]. The median time between diagnosis of CMV retinitis and onset of CME was 11.5 months (range 5-24). Development of CME was associated with significant visual loss: acuity ranged from 0.05 to 0.7 when CME was first noticed, compared to 0.8-1.25 at diagnosis of CMV retinitis. Duration of inflammation, size or zone of retinal necrosis did not favor the development of CME, neither did the antiviral therapy. A weak correlation of CME development and immune status (expressed as increase of CD4+ cells) was found. Due to systemic corticosteroids CME resolved.CONCLUSIONS: CME is a new visual threat to AIDS-patients with CMV retinitis whose immune status improved under the latest combined antiretroviral therapy. Therapy with oral corticosteroids may positively influence this condition.

Allozyme analyses have suggested that Neotropical orchid bee (Euglossini) pollinators are vulnerable because of putative high frequencies of diploid males, a result of loss of sex allele diversity in small hymenopteran populations with single locus complementary sex determination. Our analysis of 1010 males from 27 species of euglossine bees sampled across the Neotropics at 2–11 polymorphic microsatellite loci revealed only five diploid males at an overall frequency of 0.005 (95% CIs 0.002–0.010); errors through genetic nondetection of diploid males were likely small. In contrast to allozyme-based studies, we detected very weak or insignificant population genetic structure, even for a pair of populations >500 km apart, possibly accounting for low diploid male frequencies. Technical flaws in previous allozyme-based analyses have probably led to considerable overestimation of diploid male production in orchid bees. Other factors may have a more immediate impact on population persistence than the genetic load imposed by diploid males on these important Neotropical pollinators.

GP5 and M, the major membrane proteins of porcine reproductive and respiratory syndrome virus (PRRSV), are the driving force for virus budding and a target for antibodies. We studied co-translational processing of GP5 from an European PRRSV-1 strain. Using mass spectrometry, we show that in virus particles of a Lelystad variant, the signal peptide of GP5 was absent due to cleavage between glycine-34 and asparagine-35. This cleavage site removes an epitope for a neutralizing monoclonal antibody, but leaves intact another epitope recognized by neutralizing pig sera. Upon ectopic expression of this GP5 in cells, signal peptide cleavage was however inefficient. Complete cleavage occurred when cysteine-24 was changed to proline or an unused glycosylation site involving asparagine-35 was mutated. Insertion of proline at position 24 also caused carbohydrate attachment to asparagine-35. Glycosylation sites introduced downstream of residue 35 were used, but did not inhibit signal peptide processing. Co-expression of the M protein rescued this processing defect in GP5, suggesting a novel function of M towards GP5. We speculate that a complex interplay of the co-translational modifications of GP5 affect the N-terminal structure of the mature proteins and hence its antigenicity.

The objective of the present study was to enhance solubility and bioavailability of itraconazole by a combined use of membrane emulsification and spray drying solidification technique. A shirasu-porous-glass (SPG) membrane with a mean pore size of 2.5 ìm was used to produce monodispersed microemulsions of itraconazole consisting of methylene chloride as the dispersed phase, a mixture of Transcutol HP and Span 20 as a stabilizer, and dextran as solid carrier dissolved in water as the continuous phase. The dispersed phase permeated through the SPG membrane into the continuous phase at an agitator speed of 150 rpm, a feed pressure of 15 kPa and a continuous phase temperature of 25°C and the resultant emulsion was solidified using spray-drying technique. Solid state characterizations of the solid emulsion showed that the crystal state of itraconazole in solid emulsion was converted from crystalline to amorphous form. The solid emulsion of itraconazole displayed a significant increase in the dissolution rate than that of pure itraconazole. Furthermore, the solid emulsion after oral administration gave about eight-fold higher AUC and about ten-fold higher C(max) in rats than pure itraconazole powder (p<0.05), indicating this formulation greatly improved the oral bioavailability of drug in rats. Thus, these results demonstrated that the SPG membrane emulsification system combined with spray-drying technique could be used as a promising technique to develop solid formulation of itraconazole with enhanced solubility and bioavailability.

Morchella (morel) includes prized edible and medical mushrooms in the world. Since 2012, commercial cultivation of morels in the field has developed rapidly in China. However, coupled with the rapid expansion of morel cultivation, diseases have been become serious threats to morel production. White mold is one of the most serious diseases on cultivated morels. This study aimed to confirm this pathogen by following Koch's postulates, and to identify it using molecular evidence. Our results indicated that healthy Morchella fruiting bodies inoculated with Paecilomyces sp. isolates produced typical white mold symptoms, and the internal transcribed spacer sequences of the Paecilomyces sp. were 99% similar to that recovered from an epitype of Paecilomyces penicillatus. Therefore, P. penicillatus was considered to be the causative agent of white mold. White mold occurred from the initial harvest to the storage and preservation process, and it produced white mold-like symptoms on the caps and stripes of Morchella. This is the first time that white mold has been reported on cultivated Morchella.

We used polyacrylamide gel electrophoresis to examine the regulation and adenylylation states of glutamine synthetases (GSs) from Escherichia coli (GS(E)) and Klebsiella aerogenes (GS(K)). In gels containing sodium dodecyl sulfate (SDS), we found that GS(K) had a mobility which differed significantly from that of GS(E). In addition, for both GS(K) and GS(E), adenylylated subunits (GS(K)-adenosine 5'-monophosphate [AMP] and GS(E)-AMP) had lesser mobilities in SDS gels than did the corresponding non-adenylylated subunits. The order of mobilities was GS(K)-AMP < GS(K) < GS(E)-AMP < GS(E). We were able to detect these mobility differences with purified and partially purified preparations of GS, crude cell extracts, and whole cell lysates. SDS gel electrophoresis thus provided a means of estimating the adenylylation state and the quantity of GS present independent of enzymatic activity measurements and of determining the strain origin. Using SDS gels, we showed that: (i) the constitutively produced GS in strains carrying the glnA4 allele was mostly adenylylated, (ii) the GS-like polypeptide produced by strains carrying the glnA51 allele was indistinguishable from wild-type GS(K), and (iii) strains carrying the glnA10 allele contained no polypeptide having the mobility of GS(K) or GS(K)-AMP. Using native polyacrylamide gels, we detected the increased amount of dodecameric GS present in cells grown under nitrogen limitation compared with cells grown under conditions of nitrogen excess. In native gels there was neither a significant difference in the mobilities of adenylylated and non-adenylylated GSs nor a GS-like protein in cells carrying the glnA10 allele.