A comparison of adeB gene expression levels under conditions of induced resistance by different drugs in vitro in Acinetobacter baumannii.
The present study was designed to analyze the difference of the adeB gene expression levels under conditions of induced resistance and to verify the significance of the AdeABC efflux pump in the in vitro induction of Acinetobacter baumannii (A. baumannii). Strains were isolated from blood or drainage samples in the First Hospital of China Medical University. The minimum inhibitory concentration (MIC) was determined by a broth microdilution method. In addition, an in vitro induction of drug-resistance tests was confirmed with the three drugs. The expression level of the adeB was determined by quantitative polymerase chain reaction (qPCR), and there were 19, 17 and 25 strains isolated from this test that were susceptible to amikacin, netilmicin and imipenem, respectively. Furthermore, 11, 15 and 8 stably resistant strains generated against amikacin, netilmicin and imipenem, respectively, by in vitro induction. The MIC values of all the imipenem-sensitive and imipenem-resistant strains decreased by no more than 2-fold under carbonyl cyanide 3-chlorophenylhydrazone (CCCP)-containing conditions. The MIC values of 10 amikacin-resistant and 14 netilmicin-resistant strains decreased 4-fold or more under CCCP-containing conditions. Furthermore, qPCR revealed that none of the imipenem-sensitive or the imipenem-resistant strains expressed the adeB gene, but there were 10 amikacin-resistant and 5 netilmicin-resistant strains that expressed the adeB gene. In conclusion, the present study confirmed that in vitro induction of drug-resistance tests in sensitive A. baumannii strains could produce drug-resistance following long-term exposure to low doses of antibacterial drugs. In addition, amikacin and netilmicin are more prone to producing drug-resistance compared to imipenem in vitro. The results of the efflux phenotype indicate the possible application of CCCP in the inhibition of the efflux system in some drugs. The inconsistency between efflux phenotype and qPCR of adeB indicates that other mechanisms may also be included in the induction of drug-resistance that work with the active mechanism in order to increase drug resistance to common clinically-used antimicrobial drugs.


Intractable obstructive shock as a result of isolated cardiac metastases: a case report.
Cardiac metastases are uncommon and difficult to diagnose clinically; thus, they are most often found only at autopsy. Here we present a case of isolated right atrial cardiac metastasis found 7 weeks after the resection of the primary tumor, which was an adenocarcinoma of the lung. The patient presented with intractable obstructive shock, caused by a ball-valve effect of the atrial lesion that prevented forward blood flow from the right atrium. Computed tomography (CT) scans and echocardiograms failed to detect the lesion, and the patient died 2 weeks later. An autopsy revealed a large, isolated right atrial metastatic adenocarcinoma.


Medical Researchers' Ancillary Care Obligations: The Relationship-Based Approach.
In this article, I provide a new account of the basis of medical researchers' ancillary care obligations. Ancillary care in medical research, or medical care that research participants need but that is not required for the validity or safety of a study or to redress research injuries, is a topic that has drawn increasing attention in research ethics over the last ten years. My view, the relationship-based approach, improves on the main existing theory, Richardson and Belsky's 'partial-entrustment model', by avoiding its problematic restriction on the scope of health needs for which researchers could be obligated to provide ancillary care. Instead, it grounds ancillary care obligations in a wide range of morally relevant features of the researcher-participant relationship, including the level of engagement between researchers and participants, and weighs these factors against each other. I argue that the level of engagement, that is, the duration and intensity of interactions, between researchers and participants matters for ancillary care because of its connection to the meaningfulness of a relationship, and I suggest that other morally relevant features can be grounded in researchers' role obligations.


Communicating indeterminate HIV Western blot test results to clients: an observational study of three community testing sites.
This study examined HIV test counselors' beliefs and practices regarding communicating indeterminate HIV test results to clients. There are many reasons for an indeterminate HIV antibody/ Western blot assay including early HIV infection, infection with other contagious diseases, autoimmune diseases, and second or subsequent pregnancies in women. Field observations of three community HIV testing sites and semistructured interviews with 16 HIV test counselors were conducted from August 2002 through June 2003. Counselors defined an indeterminate test result in different ways. They also presented several different theories of causation, yet failed to mention two potential causes of an indeterminate result, association with large animals and infection with other (non-HIV-1) HIV strains. Counselors' understanding of the meaning of an indeterminate HIV test result varied a great deal. Some counselors believed that an indeterminate result truly was ambiguous while others believed it was indicative of being either HIV-positive or HIV-negative. Counselors' primary advice to clients who received an indeterminate result was to retest immediately without a waiting period. Counselors reported that communicating indeterminate test results to clients was a challenging task because clients who received such a result were confused, angry, and/or mistrustful. Results highlight the need for further research to provide reliable estimates of the extensiveness of indeterminate HIV test results and to further assess the nature of counselor and client responses to indeterminate test results.


Reactive arthritis induced by Vibrio parahaemolyticus.
We describe a case of reactive arthritis after Vibrio parahaemolyticus infection. V. parahaemolyticus is the principal bacteria involved in the causation of foodborne gastroenterocolitis outbreaks in Japan. Several bacteria have been recognized as being involved in the development of reactive arthritis and this is the first report of reactive arthritis induced by V. parahaemolyticus.


Meta-analysis diagnostic accuracy of SNP-based pathogenicity detection tools: a case of UTG1A1 gene mutations.
Crigler-Najjar syndrome (CNS) type I and type II are usually inherited as autosomal recessive conditions that result from mutations in the UGT1A1 gene. The main objective of the present review is to summarize results of all available evidence on the accuracy of SNP-based pathogenicity detection tools compared to published clinical result for the prediction of in nsSNPs that leads to disease using prediction performance method. A comprehensive search was performed to find all mutations related to CNS. Database searches included dbSNP, SNPdbe, HGMD, Swissvar, ensemble, and OMIM. All the mutation related to CNS was extracted. The pathogenicity prediction was done using SNP-based pathogenicity detection tools include SIFT, PHD-SNP, PolyPhen2, fathmm, Provean, and Mutpred. Overall, 59 different SNPs related to missense mutations in the UGT1A1 gene, were reviewed. Comparing the diagnostic OR, PolyPhen2 and Mutpred have the highest detection 4.983 (95% CI: 1.24 - 20.02) in both, following by SIFT (diagnostic OR: 3.25, 95% CI: 1.07 - 9.83). The highest MCC of SNP-based pathogenicity detection tools, was belong to SIFT (34.19%) followed by Provean, PolyPhen2, and Mutpred (29.99%, 29.89%, and 29.89%, respectively). Hence the highest SNP-based pathogenicity detection tools ACC, was fit to SIFT (62.71%) followed by PolyPhen2, and Mutpred (61.02%, in both). Our results suggest that some of the well-established SNP-based pathogenicity detection tools can appropriately reflect the role of a disease-associated SNP in both local and global structures.


Experience and Reporting of Postnatal Depression Across Cultures: A Comparison Using Anchoring Vignettes of Mothers in the UK and India.
Postnatal mental health is often assessed using self-assessment questionnaires in epidemiological research. Differences in response style, influenced by language, culture and experience, may mean that the same response may not have the same meaning in different settings. These differences need to be identified and accounted for in cross-cultural comparisons. We describe the development and application of anchoring vignettes to investigate the cross-cultural functioning of the Edinburgh Postnatal Depression Scale (EPDS) in urban community samples in India (n = 828) and the UK (n = 549), alongside a UK calibration sample (n = 226). Participants completed the EPDS and anchoring vignettes when their children were 12-24 months old. In an unadjusted item-response theory model, UK mothers reported higher depressive symptoms than Indian mothers (d = .48, 95% CI: .358, .599). Following adjustment for differences in response style, these positions were reversed (d = -.25, 95% CI: -.391, -.103). Response styles vary between India and the UK, indicating a need to take these differences into account when making cross-cultural comparisons. Anchoring vignettes offer a valid and feasible method for global data harmonisation.


Prevalence of agenesis of frontal sinus in human skulls with metopism.
The frontal bone is an anatomical structure of the skull separated by the metopic suture in the childhood. The scientific literature indicates that metopic suture consolidates with closure in the early stages of life. Metopism is the term used to describe a metopic suture that persists up to the adulthood. Persistent metopic suture is associated potentially with the agenesis of the frontal sinus. To investigate the prevalence of absent frontal sinuses in dry skulls with metopism. The present study was performed after the approval of the local Committee of Ethics in Research. The sample consisted of dry skulls (n=245), aging between 17 and 50 years old, of the Forensic Medical Institute of Goiânia, Brazil. The skulls underwent anthropological exam in the search for metopism. Radiographic exam was performed in the skulls with metopism to verify the presence or absence of the frontal sinus. The radiographic assessment was performed with a Mobile DaRt Evolution device (Shimadzu, Kyoto, Japan) with protocol set in 64 kV and 16 mA). From the 245 dry skulls, 17 presented metopism. The length of the metopic suture in the skulls, considering the distances between nasio and bregma craniometric landmarks, ranged between 114 mm and 137 mm. Radiographic exams were performed on 16 skulls (one skull was not analysed radiographically because of extensive destruction). Only one skull (6.25%) had the frontal sinus absent. Besides the agenesis, the present study also found four (12.5%) skulls with aplasia and eight (25.0%) hyperplasia of the frontal sinus in dry skulls with metopism. The present study found a low prevalence rate of the agenesis of frontal sinuses in dry skulls with metopism.


Evaluating the effects of a risk-adapted screening program for familial colorectal cancer in individuals between 25 and 50 years of age: study protocol for the prospective population-based intervention study FARKOR.
Colorectal cancer (CRC) is the second most common malignant disease and the second most common cause of cancer death in Germany. Official CRC screening starts at age 50. As there is evidence that individuals with a family history of CRC have an increased risk of developing CRC before age 50, there are recommendations to start screening for this group earlier. This study aims to evaluate the clinical and economic effects of a risk-adapted screening program for CRC in individuals between 25 and 50 years of age with potentially increased familial CRC risk. FARKOR (Familiäres Risiko für das Kolorektale Karzinom) is a population-based prospective intervention study. All members of cooperating statutory health insurance companies between 25 and 50 years of age living in a model region in Germany (federal state of Bavaria, 3.5 million inhabitants in this age group) can participate in the program between October 2018 and March 2020. Recruitment takes place through physicians and through a public campaign. Additionally, insurances contact recently diagnosed CRC patients in order to encourage their relatives to participate in the program. Physicians assess a participant's familial history of CRC using a short questionnaire. All participants with a family history of CRC are invited to a shared decision making process to decide on further screening options consisting of either undergoing an immunological test for fecal occult blood or colonoscopy. Comprehensive data collection based on self-reported lifestyle information, medical documentation and health administrative databases accompanies the screening program. Longterm benefits, harms and the cost-effectiveness of the risk-adapted CRC screening program will be assessed by decision analytic modeling. The data collected in this study will add important pieces of information that are still missing in the evaluation of the effects and the cost-effectiveness of a risk-adapted CRC screening strategy for individuals under 50 years of age. German Clinical Trials Register, DRKS-IDDRKS00015097.


Methionine restriction for improving progeria: another autophagy-inducing anti-aging strategy?
Methionine restriction, i.e., a partial depletion of the essential sulfur amino acid methionine from nutrition, extends lifespan in model organisms including yeast, nematodes, mice and rats. Recent results indicate that this strategy also prolongs health span and longevity in 2 short-lived strains of mice (with the LmnaG609G/G609G or zmpste24-/- genotypes) that represent animal models of Hutchinson-Gilford progeria syndrome (HGPS). The beneficial effects of methionine restriction on HGPS could be linked to reduced inflammation, and improved DNA stability, as well as the normalization of lipid and bile acid metabolism. Previous work has established that behavioral, nutritional, pharmacological and genetic manipulations that extend longevity in model organisms are only efficient if they induce increased autophagic flux. Methionine restriction extends lifespan in Saccharomyces cerevisiae in an Atg5- and Atg7-dependent fashion, supporting the notion that methionine restriction may indeed mediate its antiaging effects through the induction of macroautophagy/autophagy as well. Based on these findings, we speculate that autophagy might constitute an actionable therapeutic target to treat progeroid syndromes.


In vitro effects of 4-chloro-3-ethylphenol in skeletal muscle preparations from malignant hyperthermia susceptible and normal swine.
The in vitro contracture test with halothane and caffeine is the current gold standard for diagnosis of malignant hyperthermia. This test has a sensitivity of 99.0% but a specificity of only 93.6%. Therefore, an alternative drug is desirable which distinguishes between malignant hyperthermia-susceptible and malignant hyperthermia-normal subjects with a higher specificity and sensitivity. 4-chloro-3-ethylphenol has recently been shown to trigger Ca2+-induced Ca2+-release in skeletal muscle terminal cisternae and to increase the myoplasmic free Ca2+ concentration in skeletal muscle fibres. The purpose of this study was to investigate the ability of 4-chloro-3-ethylphenol to distinguish between malignant hyperthermia-susceptible and malignant hyperthermia-normal porcine muscle specimen in the in vitro contracture test. Ten malignant hyperthermia-susceptible and 14 malignant hyperthermia-normal swine were anaesthetized and muscle biopsies were taken. For the in vitro contracture test muscle specimens were exposed to cumulative concentrations of 4-chloro-3-ethylphenol (12.5 to 200 micromol L(-1)). 4-chloro-3-ethylphenol produced contractures in a concentration-dependent manner in the malignant hyperthermia-susceptible muscle bundles. In contrast, cumulative 4-chloro-3-ethylphenol did not generate contractures in malignant hyperthermia-normal specimens. Contractures were significantly greater (P < 0.05) in the malignant hyperthermia-susceptible compared to the malignant hyperthermia-normal preparations in all 4-chloro-3-ethylphenol concentration steps from 50 micromol L(-1) to 200 micromol L(-1). There was no overlap between the two groups above a concentration of 75 micromol L(-1) in cumulative 4-chloro-3-ethylphenol in vitro contracture tests. It remains to be verified whether an in vitro contracture test with 4-chloro-3-ethylphenol can also discriminate between malignant hyperthermia-susceptible and malignant hyperthermia-normal humans. Since no prior tested agent revealed a clear differentiation in contracture development without overlap, the 4-chloro-3-ethylphenol test might be a promising new approach to the diagnosis of malignant hyperthermia.


A transparent two-dimensional in situ beam-position and profile monitor for synchrotron X-ray beamlines.
A compact, inexpensive and easy-to-construct two-dimensional in situ beam-position and profile monitor for synchrotron X-ray beamlines is presented. The device is based on the collection of spatially resolved scattered radiation from a polyimide foil. The X-ray beam passes through a foil placed in the path of the beam, which absorbs no more than 3% of the beam at 12 keV. The scattered radiation is collected at an angle of 90 degrees through a collimator located below the foil onto a CCD sensor. The device was tested on bending-magnet beamline BM26 at the ESRF synchrotron radiation source and has a positional sensitivity better than 10 microm with a large working range of 25 mm x 25 mm. Although the device is optimized for use in the range 10-12 keV, it can easily be modified for use with higher-energy beams by using a suitably chosen scattering foil.


Australian health policy on access to medical care for refugees and asylum seekers.
Since the tightening of Australian policy for protection visa applicants began in the 1990s, access to health care has been increasingly restricted to asylum seekers on a range of different visa types. This paper summarises those legislative changes and discusses their implications for health policy relating to refugees and asylum seekers in Australia. Of particular concern are asylum seekers on Bridging Visas with no work rights and no access to Medicare. The paper examines several key questions: What is the current state of play, in terms of health screening and medical care policies, for asylum seekers and refugees? Relatedly, how has current policy changed from that of the past? How does Australia compare with other countries in relation to health policy for asylum seekers and refugees? These questions are addressed with the aim of providing a clear description of the current situation concerning Australian health policy on access to medical care for asylum seekers and refugees. Issues concerning lack of access to appropriate health care and related services are raised, ethical and practical issues are explored, and current policy gaps are investigated.


FIRST REPORTS OF LIGULA INTESTINALIS AND A SCHISTOCEPHALUS SP. INFECTING SMALL-BODIED FISH IN NEW BRUNSWICK, CANADA.
Morphological characteristics and DNA sequencing were used to identify plerocercoids of a Schistocephalus sp. infecting slimy sculpin (Cottus cognatus) from northern New Brunswick and plerocercoids of Ligula intestinalis infecting blacknose dace (Rhinichthys atratulus) in Fundy National Park (FNP, New Brunswick). To our knowledge, no previous publications documented either cestode from New Brunswick, Canada. Blacknose dace represent a new host record for L. intestinalis. Identifications were made based on the presence or absence of segmentation and sequencing partial nicotinamide adenine dinucleotide dehydrogenase subunit 1 (ND1; mitochondrial DNA) and/or partial cytochrome c oxidase subunit 1 (COI; mitochondrial DNA). Plerocercoids from blacknose dace in FNP were identified as Ligula intestinalis based on >99% nucleotide identity with COI for this species in the NCBI GenBank database. Plerocercoids in slimy sculpin from northern New Brunswick were identified as a Schistocephalus sp. based on high nucleotide identity with congenerics in the NCBI GenBank database. The absence of GenBank entries with sufficient high percent identity to our specimens, and potential species hybrids in this genus, prevents species-level identification of Schistocephalus sp. plerocercoids currently. The absence of previous documentation of these cestodes might reflect recent environmental change promoting the transmission of these parasites that can modulate host fish behavior, induce sterility of host fishes, and contribute to epizootics.


The development and integrity of equine pre-antral follicles cultured in vitro with follicle-stimulating hormone (FSH) supplementation.
This study investigated the effects of different concentrations of FSH (10, 50, 100 and 200 ng/ml) in supplemented MEM+ on the development of equine pre-antral follicles that were cultured in vitro for 2 or 6 days. The ovaries (n = 5) from mares in seasonal anoestrus were collected from a local abattoir. Ten ovarian tissue fragments of approximately 3 × 3 × 1 mm were obtained from each animal. The fragments were cultured in situ for 2 days (D2) or 6 days (D6) in MEM+ or MEM+ supplemented with FSH at four different concentrations, establishing the following 11 groups: control (D0); MEM + (D2); MEM + (D6); MEM + 10 ng/ml of FSH (D2); MEM + 10 ng/ml of FSH (D6); MEM + 50 ng/ml of FSH (D2); MEM + 50 ng/ml of FSH (D6); MEM + 100 ng/ml of FSH (D2); MEM + 100 ng/ml of FSH (D6); MEM + 200 ng/ml of FSH (D2); and MEM + 200 ng/ml of FSH (D6). Follicles were observed in only 9.65% (388 of 4,018) of the histological sections. Of the 861 follicles evaluated, 488 were in the primordial stage, and 373 were in various developmental stages; 59.7% were morphologically normal. Regarding the integrity of the pre-antral follicles, the groups with 100 ng/ml FSH of 2-days culture as well as 50, 100 and 200 ng/ml FSH of 6-days culture provided the best results. In conclusion, the in vitro culture of abattoir-derived equine ovarian fragments presented better morphological integrity when supplemented with FSH for 6 days, in comparison with the MEM culture group. However, no clear effects were observed with FSH regarding the promotion of activation from a primordial to a developing follicle.


Application of fungal chitosan incorporated with pomegranate peel extract as edible coating for microbiological, chemical and sensorial quality enhancement of Nile tilapia fillets.
Fish are generous sources for providing man with his essential nutritional requirements, but the extreme susceptibility to quality deterioration hinders their optimal usage and storage. Natural derivatives are always the perfect alternatives for food preservation. The application of fungal chitosan (Ch), from Aspergillus niger, and pomegranate peel extract (PPE), in coating films for Nile tilapia (Oreochromis niloticus) fillets preservation and maintaining their microbiological, chemical and sensorial quality during cooled storage at 4°C for 30days, was investigated. Fish fillet were coated with Ch (2%) and combined Ch+PPE, at PPE percentages of 0.5, 1.0, 1.5 and 2.0%. Fillets coating resulted in sharp decrease of the entire microbial counts during storage; the increased concentrations from PPE strengthened coating film antimicrobial activity. Additionally, fillets coating could retard the chemical spoilage parameters increasing, i.e. nitrogen volatile base (TVB-N), peroxide value (PV) and reactive substances of thiobarbituric acid (TBARS), during storage period. The sensory evaluation indicated higher preferences for the odor, texture, color and overall quality of coated samples. Fish fillets coating with Ch and Ch+PPE could be recommended for shelf life extension and maintaining the microbiological, chemical and sensorial quality through the application of safe preservatives from natural origins.


An investigation of factors related to intoxicated driving behaviors among youth.
This study assessed the prevalence of driving under the influence of alcohol and marijuana among a sample of 18 and 21 year olds and examined the across-time relationships between intoxicated driving and consumption, risk-taking/impulsive orientation, negative intrapersonal state, stress and use of alcohol and other drugs to cope with problems. Self-report data were collected from 556 men and women, ages 18 and 21, at two points in time. The data indicated that at least a minimum level of drinking and driving, as well as smoking marijuana and driving, is engaged in at least once for the majority of youth. Correlations between eight driving behaviors and consumption variables indicated that frequency of substance use was strongly related to frequency of driving while intoxicated (DWI). Regression analyses revealed that coping use of substances was the strongest predictor of driving under the influence. A path model examining the effect of stress, negative states and risk-taking orientations (T1) on driving under the influence as mediated through coping use (T2) was tested. Results showed that risk-taking orientation was the strongest predictor of DWI, both directly and indirectly (as mediated through coping use). Findings suggest that impaired driving may be part of a global syndrome of risk-taking behavior and is an activity engaged in most often by those who frequently use alcohol and other drugs to cope with problems.


[Genotype and phenotype correlation of phenylalanine hydroxylase deficiency among patients from Henan].
To delineate the mutation spectrum of phenylalanine hydroxylase (PAH) gene among patients affected with phenylalanine hydroxylase deficiency (PAHD) in Henan Province of China, and to explore the correlation between the genotype and the phenotype. A total of 155 affected children were recruited. Potential mutation of the PAH gene were analyzed by direct sequencing. The genotype-phenotype correlation was analyzed by matching the expected and observed phenotypes. Over 72 mutations and 108 genotypes have been identified. There were 7 homozygous mutations, including 1 case with EX6-96A>G/EX6-96A>G, 1 with R241C/R241C, 1 with R413P/R413P, and 4 with R243Q/R243Q. Among these, 6 patients have presented classic PKU phenotypes, except for a R241C/R241C genotype which has led to mild PKU. In 104 patients carrying compound PAH mutations, 52 were classic, 34 were mild and 39 had mild HPA. Patients who were heterozygous for EX6-96A>G/R241C, R243Q/A434D, EX6-96A>G/R413P and EX6-96A>G/ R241C were found with both the classic PKU and mild PKU phenotypes. Common mutations associated with mild HPA have included R53H, R243Q, V399V and H107R. The common mutations associated with mild PKU included R243Q, R241C, EX6-96A>G, and IVS4-1G>A. The prevalent mutations in classic PKU were R243Q, EX6-96A>G and V399V. The consistency between prediction of the biochemical genotype and observed phenotype was 77.78%, especially in classic PKU, the consistency was up to 82.14%. Significant correlations were disclosed between pretreatment levels of phenylalanine and AV sum (r=-0.6729, P < 0.01). The mutation spectrum of PAH gene in Henan seems to differ from that of other regions. Independent assortment of mutant alleles may result in a complex genotype-phenotype correlation, but the genotypes of PAHD patients have correlated with the phenotype.


Coexistence of thrombotic thrombocytopenic purpura and immune thrombocytopenic purpura in an Asian woman: a case report.
A 33-year-old Chinese woman with a history of immune thrombocytopenic purpura presented with heavy menstrual bleeding. She was found to have thrombocytopenia, plasma ADAMTS13 activity of 0%, and positivity for the plasma ADAMTS13 inhibitor. She was diagnosed with the coexistence of thrombotic thrombocytopenic purpura and immune thrombocytopenic purpura. The patient was treated by plasmapheresis, a glucocorticoid, and rituximab. Her platelet level returned to normal, and she was discharged 28 days after admission. The number of plasmapheresis sessions and the timing of rituximab administration may be the key aspects of management of patients with thrombotic thrombocytopenic purpura who have underlying immune dysfunction caused by diseases such as immune thrombocytopenic purpura.


Hypoglycemia complicating dehydration due to acute gastroenteritis.
To identify clinical variables predictive of hypoglycemia in dehydrated children with acute gastroenteritis (AGE), clinical variables were recorded for dehydrated children younger than 5 years old with AGE before measuring serum glucose. One hundred and eighty-four children were enrolled; 62 with, and 122 without, hypoglycemia. Multivariate analysis identified variables associated with hypoglycemia. Female gender, neurologic symptoms of hypoglycemia, and a greater amount of vomiting vs. diarrhea were significantly associated with hypoglycemia. Clinical variables do not have adequate sensitivity and specificity to accurately predict which children with AGE have hypoglycemia. Glucose determination should be considered for these patients.


Assessment of gelatinases (MMP-2 and MMP-9) by gelatin zymography.
Gelatin zymography is a simple yet powerful method to detect proteolytic enzymes capable of degrading gelatin from various biological sources. It is particularly useful for the assessment of two key members of the matrix metalloproteinase family, MMP-2 (gelatinase A) and MMP-9 (gelatinase B), due to their potent gelatin-degrading activity. This polyacrylamide gel electrophoresis-based method can provide a reliable assessment of the type of gelatinase, relative amount, and activation status (latent, compared with active enzyme forms) in cultured cells, tissues, and biological fluids. The method can be used to investigate factors that regulate gelatinase expression and modulate zymogen activation in experimental systems. The system provides information on the pattern of gelatinase expression and activation in human cancer tissues and how this relates to cancer progression. Interpretation of the data obtained in gelatin zymography requires a thorough understanding of the principles and pitfalls of the technique; this is particularly important when evaluating enzyme levels and the presence of active gelatinase species. If properly used, gelatin zymography is an excellent tool for the study of gelatinases in biological systems.


New developments in probing and targeting protein acylation in malaria, leishmaniasis and African sleeping sickness.
Infections by protozoan parasites, such as Plasmodium falciparum or Leishmania donovani, have a significant health, social and economic impact and threaten billions of people living in tropical and sub-tropical regions of developing countries worldwide. The increasing range of parasite strains resistant to frontline therapeutics makes the identification of novel drug targets and the development of corresponding inhibitors vital. Post-translational modifications (PTMs) are important modulators of biology and inhibition of protein lipidation has emerged as a promising therapeutic strategy for treatment of parasitic diseases. In this review we summarize the latest insights into protein lipidation in protozoan parasites. We discuss how recent chemical proteomic approaches have delivered the first global overviews of protein lipidation in these organisms, contributing to our understanding of the role of this PTM in critical metabolic and cellular functions. Additionally, we highlight the development of new small molecule inhibitors to target parasite acyl transferases.


A look behind closed doors: interaction of persistent viruses with dendritic cells.
Persistent infections with HIV, hepatitis B virus and hepatitis C virus are major causes of morbidity and mortality worldwide. As sentinels of the immune system, dendritic cells (DCs) are crucial for the generation of protective antiviral immunity. Recent advances in our understanding of the role of DCs during infection with these viruses provide insights into the mechanisms used by these viruses to exploit DC function and evade innate and adaptive immunity. In this Review we highlight the current knowledge about the interaction between DCs and these viruses and the underlying mechanisms that might influence the outcome of viral infections.


Sources of social support related to single-parent functioning.
Factors related to successful functioning of single parents have not been identified. In this study, relationships among sources of social support and criterion measures of functioning were examined. To isolate the role of social support, a population was selected that was relatively homogeneous on variables considered important in coping with single parenthood: economic status, educational or vocational opportunities, access to day-care, and length of time as a single parent. A check list and structured interview were administered to 30 unmarried women to determine their sources of social support. Ratings of parenting adequacy and of the preschool child's emotional development and behavior by three professional staff members at the day-care centers were used as criterion measures of functioning. Based on the staff ratings, the adequately functioning group was distinquished from the inadequate one. Significant correlations were found between these staff ratings and one social support variable: support from talking with network the members. Key sources of social support were identified through discriminant analyses.


Simultaneous Changes in Sleep, qEEG, Physiology, Behaviour and Neurochemistry in Rats Exposed to Repeated Social Defeat Stress.
Depression is a heterogeneous disorder characterized by alterations at psychological, behavioural, physiological, neurophysiological, and neurochemical levels. Social stress is a prevalent stress in man, and the repeated social defeat stress model in rats has been proposed as being the rodent equivalent to loss of control, which in subordinate animals produces alterations that resemble several of the cardinal symptoms found in depressed patients. Here, rats followed a resident-intruder protocol for 4 consecutive days during which behavioural, physiological, and electroencephalographic (EEG) parameters were simultaneously monitored in subordinate rats. On day 5, prefrontal dopamine (DA) and hippocampal serotonin (5-HT) as well as corticosterone were measured in submissive rats that had visual, acoustic, and olfactory (but no physical) contact with a dominant, resident conspecific rat. Socially defeated rats demonstrated increases in ultrasonic vocalizations (20-25 KHz), freezing, submissive defensive behaviour, inactivity, and haemodynamic response, while decreases were found in repetitive grooming behaviour and body weight. Additionally, alterations in the sleep-wake architecture were associated with reduced active waking, enhanced light sleep, and increased frequency of transitions from light sleep to quiet wakefulness, indicating sleep instability. Moreover, the attenuation of EEG power over the frequency range of 4.2-30 Hz, associated with a sharp transient increase in delta oscillations, appeared to reflect increased brain activity and metabolism in subordinate animals. These EEG changes were synchronous with a marked increase in body temperature and a decrease in locomotor activity. Furthermore, psychosocial stress consistently increased 5-HT, DA, and corticosterone levels. The increased levels of cortical DA and hippocampal 5-HT during social threat may reflect a coping mechanism to promote alertness and psychological adaptation to provocative and threatening stimuli. These neurophysiological changes are hypothesized to be the consequence of dynamics in monoamine systems, which could be useful markers for disease progression in the aetiology of depression.


The antidiuretic action of 1-deamino-8-D-arginine vasopressin (DDAVP) in man.
1. After administration of a new vasopressin analogue (DDAVP), a marked and prolonged antidiuresis occurred in 10 patients with pituitary diabetes insipidus. 2. The antidiuretic effects of single intravenous doses of 0.04--24 mug DDAVP and single intranasal doses of 5--320 mug DDAVP were investigated. Time curves of the antidiuretic responses expressed in changes of urine osmolality (Uosm) and free water clearance per 100 ml GFR (CH2O X 100/GFR) are described. 3. Maximal "peak" response was obtained after an intravenous dose of 1 mug within the first 12 hrs (Uosm was 7--800 mOsm/KgH2O). Further increase of dosage resulted only in prolongation of duration of action (up to 48 hrs) and peak ("plateau") effect (up to 24 hrs). 4. There was a linear relationship between the log dose and log osmolality of urine collected in the second 12 hours after administration of single intravenous and intranasal doses of DDAVP. 5. Comparison of the effects of 1 mug lysine-vasopressin and 1 mug DDAVP revealed only slight differences in peak effects, but extreme differences in duration of action. 6. It is concluded that in the evaluation of a synthetic vasopressin analogue the maximal antidiuretic ability and the prolongation of action have to be analysed separately.


Malignant tumours of the maxillary complex: an 18-year review.
Over an 18-year period, 147 patients with malignant tumours involving the maxillary complex were treated in a combined head and neck clinic, of whom 50 underwent surgery. There were 33 males and 17 females, the mean age was 57 years (range 11-87 years). The most common clinical presentations were either painful facial swelling, infraorbital anaesthesia, palatal ulceration or nasal obstruction. Preoperative investigations included EUA, biopsy and either CT or MRI scans. Of the 50 tumours, 62% were squamous cell carcinomas. Surgery consisted of either partial or total maxillectomy including craniofacial resection in nine patients. Reconstruction was by either split skin grafting or by free tissue transfer when the cranial contents had been exposed. Adjuvant radiotherapy was given in 82% of the patients. The mean follow-up was 5 years and 59% of patients are alive and disease free. The 5-year local control rate was 67%; it was greatly influenced by histological evidence of nerve invasion, local recurrence being the major cause of death. It was concluded that adequate surgical clearance, followed by planned postoperative radiotherapy, is the most effective treatment for malignant disease of the maxillary complex.


[Arthroscopic palliative resection of the rotator cuff in patients with unreconstructible lesions].
The aim of this study is to present a simple rotator cuff lesion classification that provides guidelines as to their treatment, and to evaluate the results of palliative arthroscopic resection of rotator cuff residues known as unreconstructible lesions. In addition, our therapeutic approaches were ascertained in view of their applicability to the types of lesions studied. In a five-year period (January 1, 2000 to December 31, 2004), a total of 181 arthroscopic procedures were performed on the shoulder joints of patients diagnosed with impingement or rotator cuff syndromes. In 130 cases, a tear or irritation of the rotator cuff was recorded. Rotator cuff lesions were categorized on the basis of our modification of the Gschwend classification. In 15 of the patients, in whom unreconstructible lesions were detected, arthroscopic palliative resection of rotator cuff residues was performed. The average age of these patients was 65 years, and they were followed up for 6 to 60 months. All surgery was carried out in a "beach-chair" position, either under general anesthesia or with an interscalene brachial plexus block. The arthroscope was inserted through the "soft-spot". Continuous irrigation was provided with an arthroscopic pump. In the first place, the glenohumeral joint was explored, and resection of rotator cuff residues was performed via ventral and lateral ports. The procedure was completed by subacromial decompression and partial resection of the acromion. The results were evaluated by the Constant Functional Score, as modified by us. Clinical examination was supplemented with subjective information from questionnaires provided by the patients. In a total of 130 shoulder joints with rotator cuff tears examined by arthroscopy, type I lesions were found in 90, and these were treated by arthroscopic subacromial decompression. Twenty-five type II and type III lesions underwent open rotator cuff repair and 15 type IV and type V lesions were treated by palliative arthroscopic resection of residual rotator cuff lesions, using the Apoil method. These fifteen patients were followed up for 6 to 60 months and their outcomes were evaluated. No excellent results were achieved (Constant Score, 80-100 points), but this is implicit in the nature of a palliative operation. Good (65-79 points) and satisfactory (51-64 points) results were recorded in 11 (73.3 %) and four (26.7 %) patients, respectively. No poor results were found. The average improvement in Constant scores was 21 points. A total of 130 rotator cuff lesions diagnosed arthroscopically were categorized on the basis of a modified classification system. We will continue to treat type I lesions by arthroscopic subacromial decompression, which has provided good results, as reported in our previous study. We consider the arthroscopic repair of rotator cuff tears to be an optimal procedure for type II lesions; for type III lesions we will keep using open repair surgery. The most complex problem is presented by type IV lesions. While palliative arthroscopic resection of the rotator cuff is one option, muscle transfer has also shown satisfactory outcomes, as has partial reconstruction. The use of either allografts or cadaver grafts did not give good results. Type V lesions, in our opinion, are unambiguously indicated for palliative arthroscopic resection of the rotator cuff. Their treatment by the Apoil method and detailed evaluation of the outcomes are described here; the results of this study are in agreement with those reported in the relevant international literature. Good and satisfactory results were achieved by palliative arthroscopic resection of the rotator cuff, in combination with subacromial decompression, in patients with unreconstructible lesions. The average improvement in the Constant Functional Score was 21 points. This suggests that the method can be recommended for wider use in the future. However, exact diagnosis and correct indication, i.e., type V lesion, are essential.


The military physician assistant.
The origin of the physician assistant began with medically trained military personnel. Now, 25 years later, the military has over 1,100 physician assistants working in all aspects of health care delivery. This article examines the historical perspective and discusses current and emerging issues for the utilization of military physician assistants.


[Cochlear implants in obliterated or ossified cochleas].
Our last cases of cochlear implants in obliterated or ossified cochleas are presented, and we had described the surgical technique for double-array or compressed system. In our opinion the best surgical method is the combined approach (endoaural with aticotomy view and posterior timpanotomy) for increased promontory view. This technique provides the advantage that it always preserve the posterior wall of the external ear canal, and the array is positioned easily.


Structure, Spectra, and DFT Simulation of Nickel Benzazolate Complexes with Tris(2-aminoethyl)amine Ligand.
Benzazolate complexes of Ni(II), [Ni(pbz)(tren)]ClO4 (pbz = 2-(2'-hydroxyphenyl)-benzimidazole (pbm), 1, 2-(2'-hydroxyphenyl)-benzoxazole (pbx), 2, 2-(2'-hydroxyphenyl)-benzothiazole (pbt), 3; tren = tris(2-aminoethyl)amine), are prepared by self-assembly reaction and structurally characterized. Theoretical DFT simulations are carried out to reproduce the features of their crystal structures and their spectroscopic and photophysic properties. The three complexes are moderately luminescent at room temperature both in acetonitrile solution and in the solid state. The simulations indicate that the absorption spectrum is dominated by two well-defined transitions, and the electronic density concentrates in three MOs around the benzazole ligands. The Stokes shifts of the emission spectra of complexes 1-3 are determined by optimizing the electronic excited state.


Exploring visual plasticity: dietary carotenoids can change color vision in guppies (Poecilia reticulata).
Differences in color vision can play a key role in an organism's ability to perceive and interact with the environment across a broad range of taxa. Recently, species have been shown to vary in color vision across populations as a result of differences in regulatory sequence and/or plasticity of opsin gene expression. For decades, biologists have been intrigued by among-population variation in color-based mate preferences of female Trinidadian guppies. We proposed that some of this variation results from variation in color vision caused by plasticity in opsin expression. Specifically, we asked about the role of dietary carotenoid availability, because carotenoids (1) are the precursors for vitamin A, which is essential for the creation of photopigments and (2) have been linked to variation in female mate choice. We raised guppies on different carotenoid-level diets and measured opsin expression. Guppies raised on high-carotenoid diets expressed higher levels of long wavelength sensitive opsin (LWS) opsins than those raised on lower levels of carotenoids. These results suggest that dietary effects on opsin expression represent a previously unaccounted for mechanism by which ecological differences across populations could lead to mate choice differences.


Erastin inhibits porcine epidemic diarrhea virus replication in Vero cells.
Porcine epidemic diarrhea virus (PEDV), an intestinal pathogenic coronavirus, has caused significant economic losses to the swine industry worldwide. At present, there are several treatment methods, but there is still a lack of clinically effective targeted drugs, new antiviral mechanisms and drugs need to be explored. In this study, we established a model of erastin versus ferrostatin-1 treatment of Vero cells, and then detected virus proliferation and gene expression by RT-qPCR through PEDV infection experiments. We demonstrated for the first time that erastin significantly inhibited the replication of PEDV upon entry into cells; Vero treated with erastin significantly regulated the expression of three genes, NRF2, ACSL4 and GPX4, notably erastin regulated the expression of these three genes negatively correlated with the expression induced by PEDV virus infection. Since NRF2, ACSL4 and GPX4 are classical Ferroptosis genes, this study speculates that erastin may inhibit the replication of PEDV in Vero cells in part through the regulation of ferroptosis pathway, and erastin may be a potential drug for the treatment of PEDV infection.


Brief report: development of a prescription medication information webliography for consumers.
Websites offering drug information vary in coverage and quality, and most health care consumers are poorly equipped to assess the quality of internet medication information. To establish a webliography of recommended prescription medication information websites for health care consumers and providers. Drug information websites were systematically identified based on recommendations from health professionals and text-word searches of MEDLINE and Google. The resulting sample of websites was evaluated in a 2-step process. Candidate websites were first screened using inclusion/exclusion criteria representing minimum information requirements. Websites that passed the inclusion/exclusion criteria were then rated on 16 quality criteria using a 5-point scale by 3 trained judges. Website ratings were averaged, then multiplied by the corresponding importance weight of each criterion and summed to generate a total score. Websites with the highest total scores were included in the webliography. Ten websites were selected for inclusion in the webliography. The 3 highest-scoring websites were Anthem Blue Cross and Blue Shield (http://home.anthemhealth.com/topic/drugcenter), U.S. National Library of Medicine (https://www.nlm.nih.gov/medlineplus/druginformation.html), and Healthvision (http://www.yourhealthinformation.com/library/healthguide/en-us/drugguide/default.htm). Medication information websites vary widely in quality and content. The online webliography is a valuable and easily accessed tool that can be recommended by health care professionals to patients who request referral to reliable websites.


Analysis of specific binding involved in genomic packaging of the double-stranded-RNA bacteriophage phi6.
The genomes of bacteriophage phi6 and its relatives are packaged through a mechanism that involves the recognition and translocation of the three different plus-strand transcripts of the segmented double-stranded-RNA genomes into preformed polyhedral structures called procapsids or inner cores. The packaging requires the hydrolysis of nucleoside triphosphates and takes place in the order segment S-segment M, segment L. Packaging is dependent upon unique sequences of about 200 nucleotides near the 5' ends of plus-strand transcripts of the three genomic segments. It appears that P1 is the determinant of the RNA binding sites. Directed mutation of P1 was used to locate regions that are important for genomic packaging. Specific binding of RNA to the exterior of the procapsid was dependent upon ATP, and a region that showed a high level of cross-linking to phage-specific RNA was located. Antibodies to peptide sequences were prepared, and their abilities to bind to the exterior of procapsids were determined. Sites sensitive to trypsin and to factor Xa were determined as well.


Green Solvent Ethanol-Based Inks for Industrially Applicable Deposition of High-Quality Perovskite Films for Optoelectronic Device Applications.
Incontrovertibly there is an increasing demand for the development of benign inks suitable for fabrication of high-performing perovskite-based thin film functional layers. Nevertheless, most reported perovskite precursors rely on the use of highly toxic solvents such as acetonitrile, 2-methoxyethanol, dimethylformamide, and many others. Hence, there is a strong imperative for the development of novel and greener inks, which will facilitate smoother commercialization of technologies based on functional perovskite films. Therefore, four perovskite precursors are studied, some of which consist of up to 90% ethanol. All inks are developed to fulfill the requirements of a high-throughput deposition compatible with roll-to-roll techniques at room temperature, assisted by an air knife for instant solvent removal. Two of the inks are particularly suitable for the fabrication of high-quality and densely packed multi-crystalline (CH3 NH3 )PbI3 layers, as confirmed by numerous nanoscale spectroscopic and material characterization techniques. Additionally, large-area photoluminescence (PL) imaging is demonstrated to improve the quality of the deposited perovskite films, with a route to enhance deposition uniformity when upscaling for manufacture. The genuine potential of the developed greener perovskite inks is demonstrated with the fabrication of solar cells with power conversion efficiencies above 19.5%.


Regulation of cytoskeletal dynamics by redox signaling and oxidative stress: implications for neuronal development and trafficking.
A proper balance between chemical reduction and oxidation (known as redox balance) is essential for normal cellular physiology. Deregulation in the production of oxidative species leads to DNA damage, lipid peroxidation and aberrant post-translational modification of proteins, which in most cases induces injury, cell death and disease. However, physiological concentrations of oxidative species are necessary to support important cell functions, such as chemotaxis, hormone synthesis, immune response, cytoskeletal remodeling, Ca(2+) homeostasis and others. Recent evidence suggests that redox balance regulates actin and microtubule dynamics in both physiological and pathological contexts. Microtubules and actin microfilaments contain certain amino acid residues that are susceptible to oxidation, which reduces the ability of microtubules to polymerize and causes severing of actin microfilaments in neuronal and non-neuronal cells. In contrast, inhibited production of reactive oxygen species (ROS; e.g., due to NOXs) leads to aberrant actin polymerization, decreases neurite outgrowth and affects the normal development and polarization of neurons. In this review, we summarize emerging evidence suggesting that both general and specific enzymatic sources of redox species exert diverse effects on cytoskeletal dynamics. Considering the intimate relationship between cytoskeletal dynamics and trafficking, we also discuss the potential effects of redox balance on intracellular transport via regulation of the components of the microtubule and actin cytoskeleton as well as cytoskeleton-associated proteins, which may directly impact localization of proteins and vesicles across the soma, dendrites and axon of neurons.


Quantitative analysis of the distribution and content of trace metal elements in liquid samples with a three-dimensional confocal x-ray technique.
Three-dimensional (3D) confocal x-ray fluorescence analysis technology is widely used, but the quantitative analysis of elemental spatial distributions of solid samples is complicated. This paper explores a quantitative analysis method that can be applied to solid samples. Fluorescence spectra of liquid samples are obtained on a 3D confocal x-ray fluorescence spectrometer. Curves are plotted showing the relationships between the fluorescence count intensity and the mass percentage of metal ions, and the respective fitting-curve equations are determined according to the curve morphology. Fluorescence intensity as a function of the mass percentage and depth position is derived from the samples for a particular acquisition time. These data play a potential role in the subsequent quantitative analysis of unknown mass percentages of solid samples.


[The research advances of biomechanics of human knee joint ligaments].
Ligaments are the main parts which stabilize the knee joint. How to analog the ligaments in biomechanical model will affect the characteristics of the human knee dynamics and in the computation of the stress in ligaments between two bones. This symposium is aimed at the survey of the simplified method of the ligaments via mechanical parameters, and providing an exact method of constructing model.


C4b-binding protein, a regulatory component of the classical pathway of complement, is an acute-phase protein and is elevated in systemic lupus erythematosus.
A radioimmunoassay using monoclonal and polyclonal antihuman C4b-binding protein (C4BP) antibody was developed to quantitate C4BP in serum. Using the assay, the levels of C4BP in healthy individuals, in patients with systemic lupus erythematosus (SLE), and in acute-phase individuals were determined. The levels of C4BP are significantly elevated in individuals with SLE (186%; p = 0.0001) and are even higher in individuals during the acute phase (286%; p = 0.0001). To confirm whether or not individuals were in the acute-phase response, serum C-reactive protein (CRP) levels were assessed. In the acute-phase response, CRP levels were 100-fold elevated over normals, but did not correlate with increases in C4BP (r = -0.031; p = 0.899). In SLE patients, the CRP levels were significantly, but moderately, elevated (5-fold; p = 0.028). The data indicate that C4BP is an acute-phase reactant and is differentially regulated from CRP during the acute-phase response.


Normative data for two MMPI critical item sets.
Addressed normative aspects of recent MMPI critical item sets proposed by Koss and Butcher (1973) and Lachar and Wrobel (1979). Base rates from MMPI Form R for these items were determined in samples of 123 normals, 383 inpatient alcoholics, and 567 psychiatric inpatients. Although pathological groups, as expected, generally endorsed more critical items than normals, mean differences were not large. On several critical item subsets, especially in the Koss -Butcher group, normals actually scored higher than the pathological samples. Caution is suggested when critical MMPI items are used in assessments.


Unfolding homoclinic connections formed by corner intersections in piecewise-smooth maps.
The stable and unstable manifolds of an invariant set of a piecewise-smooth map are themselves piecewise-smooth. Consequently, as parameters of a piecewise-smooth map are varied, an invariant set can develop a homoclinic connection when its stable manifold intersects a non-differentiable point of its unstable manifold (or vice-versa). This is a codimension-one bifurcation analogous to a homoclinic tangency of a smooth map, referred to here as a homoclinic corner. This paper presents an unfolding of generic homoclinic corners for saddle fixed points of planar piecewise-smooth continuous maps. It is shown that a sequence of border-collision bifurcations limits to a homoclinic corner and that all nearby periodic solutions are unstable.


Synthetic Nucleic Acids and Treatment of Neurological Diseases.
The ability to control gene expression with antisense oligonucleotides (ASOs) could provide a new treatment strategy for disease. To review the use of ASOs for the treatment of neurological disorders. Articles were identified through a search of PubMed references from 2000 to 2016 for articles describing the use of ASOs to treat disease, with specific attention to neurological disease. We concentrated our review on articles pertaining to activation of frataxin expression (Friedreich's ataxia) and production of active survival motor neuron 2 (SMN2, spinal muscular atrophy). Many neurological diseases are caused by inappropriate expression of a protein. Mutations may reduce expression of a wild-type protein, and strategies to activate expression may provide therapeutic benefit. For other diseases, a mutant protein may be expressed too highly and methods that reduce mutant protein expression might form the basis for drug development. Synthetic ASOs can recognize cellular RNA and control gene expression. Antisense oligonucleotides are not a new concept, but successful clinical development has proceeded at a slow pace. Advances in ASO chemistry, biological understanding, and clinical design are making successful applications more likely. Both laboratory and clinical studies are demonstrating the potential of ASOs as a source of drugs to treat neurological disease.


Score test for detecting linkage to quantitative traits.
The two most popular methods to detect linkage of a quantitative trait to a marker are the Haseman-Elston regression method and the variance components likelihood-ratio test. In the literature, these methods are frequently compared and the relative advantages and disadvantages of each method are well known. In this article, we derive a score test for the variance component attributable to a specific quantitative trait locus and show that for sib-pairs it is mathematically equivalent to a recently proposed version of the Haseman-Elston method that optimally combines the sum squared and the difference squared of the centered phenotype values of the sibs. Because score tests and likelihood-ratio tetsts are equivalent for large sample sizes, the variance components likelihood-ratio test is also asymptotically equivalent to this optimal Haseman-Elston test. This fact gives a theoretical explanation of the empirical observation from simulation studies reporting similar power of the variance components likelihood-ratio test and the optimal Haseman-Elston method. Perhaps more importantly for practical purposes, the score test can also be extended in a natural way to support the simultaneous analysis of more than two subjects and multivariate phenotypes.


Multiple tumor types may originate from bone marrow-derived cells.
It was believed that tumors originated from the transformation of their tissue-specific stem cells. However, bone marrow-derived cells (BMDCs), which possess an unexpected degree of plasticity and often reside in other tissues, might also represent a potential source of malignancy. To study whether BMDCs play a role in the source of other tumors, BMDCs from mice were treated with 3-methycholanthrene until malignant transformation was achieved. Here we show that transformed BMDCs could form many tumor types, including epithelial tumors, neural tumors, muscular tumors, tumors of fibroblasts, blood vessel endothelial tumors, and tumors of poor differentiation in vivo. Moreover, a single transformed BMDC has the ability to self-renew, differentiate spontaneously into various types of tumor cells in vitro, express markers associated with multipotency, and form teratoma in vivo. These data suggest that multipotent cancer stem cells seemed to originate from transformed BMDCs. Conclusively, these findings reveal that BMDCs might be a source of many tumor types, even teratoma. In addition, multipotent cancer stem cells might originate from malignant transformed BMDCs.


Nutrient availability shapes methionine metabolism in p16/MTAP-deleted cells.
Codeletions of gene loci containing tumor suppressors and neighboring metabolic enzymes present an attractive synthetic dependency in cancers. However, the impact that these genetic events have on metabolic processes, which are also dependent on nutrient availability and other environmental factors, is unknown. As a proof of concept, we considered panels of cancer cells with homozygous codeletions in CDKN2a and MTAP, genes respectively encoding the commonly-deleted tumor suppressor p16 and an enzyme involved in methionine metabolism. A comparative metabolomics analysis revealed that while a metabolic signature of MTAP deletion is apparent, it is not preserved upon restriction of nutrients related to methionine metabolism. Furthermore, re-expression of MTAP exerts heterogeneous consequences on metabolism across isogenic cell pairs. Together, this study demonstrates that numerous factors, particularly nutrition, can overwhelm the effects of metabolic gene deletions on metabolism. These findings may also have relevance to drug development efforts aiming to target methionine metabolism.


Bioresorbable implants: applications in orthopaedic surgery.
There are several new and potential uses for absorbable implants. Some will provide exciting options in the future. Ongoing animal and clinical studies will help focus future areas of development.


Sequential monitoring of pigmented lesions during dabrafenib treatment: a prospective study and a literature overview.
Targeted therapies in melanoma have shown clinical benefit in incrementing the overall survival of metastatic patients. However, cutaneous adverse events have been frequently associated with these drugs. We report our experience in the management of patients treated with dabrafenib for metastatic melanoma, focusing on the monitoring of pigmented lesions. Dermatologic evaluation was performed during the first visit, at the start of each treatment and subsequently after every four weeks. Global nevi count, videodermoscopy of suspected lesions, and surgical excisions when necessary were performed at the beginning of the treatment and every fourth week. All other cutaneous adverse events (cAEs) were noted and documented. Eleven patients were included. The most important cAEs included palmo-plantar hyperkeratosis, diffuse xerosis and pigmented lesion changes. Regarding the latter, in 6 patients, especially in the first months of treatment, we observed hyperpigmentation and hyperkeratosis of the nevi, of the pigmented mucosae and, in one patient, hyperkeratotic changes on a cutaneous metastasis. Histopathology of the excised lesions showed one ex novo melanoma occurrence and benign changes to pre-existing nevi. The awareness of the importance of sequential monitoring of pigmented lesions, with particular attention to the lesions of new onset, is crucial for the best management of these complex patients.


Polymorph selection during the crystallization of softly repulsive spheres: the inverse power law potential.
Using hybrid Monte Carlo molecular simulations, we study crystallization from the melt of softly repulsive spheres interacting through an inverse power law potential. We work at fixed supercooling (i.e., at a temperature 25% below the melting temperature) and consider three systems, defined by different values for the inverse power exponent n: n = 5, n = 6.67, and n = 10. Modifying the value of n allows us to study the onset of crystallization in the domain of stability of the body-centered cubic (bcc) phase (n = 5 and n = 6.67) and in the domain of stability of the face-centered cubic (fcc) phase (n = 10). We show that, for the three systems, polymorph selection does not take place during crystal nucleation since the structure of the critical nuclei obtained for the three systems is not well defined. However, our results demonstrate that polymorph selection takes place during the growth step since growth proceeds either into the stable bcc phase for the two smaller values of n (n = 5 and n = 6.67) or into the stable fcc phase for the larger value of n (n = 10). We also show that we did not achieve complete control of polymorphism for n = 10. The growth step gives rise to either slowly growing crystallites composed of two blocks of different structures (the stable fcc form and the metastable bcc form) or rapidly growing crystallites of the metastable bcc form.


Nano-Zoo Interfacial Interaction as a Design Principle for Hybrid Soil Remediation Technology.
Using nanotechnology to remediate contaminated agricultural soil is promising but faces notable technical and economic challenges. Importantly, widely distributed soil invertebrates can potentially act as natural mobile facilitators for in situ nanoscale remediation of contaminated soil. Herein, we have drawn inspiration from nano-bio interaction and established a hybrid remediation framework using nanoscale zerovalent iron (nZVI) and nematodes for organochlorine-contaminated soil. Approximately 80% pentachlorophenol (PCP, initially 50 mg/kg) was synergistically degraded by nZVI and nematodes within 3 days. Mechanistically, exposure to nZVI stimulated the synthesis of reductive biomolecules (including collagen, glutathione, and l-cysteine) which acted as a bioreductive barrier and significantly mitigated the toxicity of PCP. At the microinterface, collagen distributed in the epidermis chelated nZVI; subsequently, l-cysteine and glutathione strongly accelerated nZVI-induced PCP dechlorination by facilitating the reductive dissolution of nZVI oxide shell and electron transfer from Fe0 core to PCP. On the basis of the interfacial interaction, an optimized soil remediation approach composed of nZVI, nematodes, and l-cysteine was established, demonstrating a 2.1-fold increase in removal efficiency with only 48.5% nZVI consumption compared with the nZVI treatment alone. This work provides a heuristic model for developing cost-efficient remediation technologies with the synergistic force of functional materials and indigenous biota, which may be widely applicable to a range of environmental contamination scenarios.


Knowledge, attitudes, and practices among health care providers regarding complementary and alternative medicine in Trinidad and Tobago.
Health care providers are often ill prepared to interact about or make acceptable conclusions on complementary and alternative medicine (CAM) despite its widespread use. We explored the knowledge, attitudes, and practices of health care providers regarding CAM. This cross-sectional study was conducted between March 1 and July 31, 2015 among health care providers working mainly in the public sector in Trinidad and Tobago. A 34-item questionnaire was distributed and used for data collection. Questionnaire data were analysed using inferential and binary logistic regression models. Response rate was 60.3% (362/600). Responders were 172 nurses, 77 doctors, 30 pharmacists, and 83 other health care providers of unnamed categories (mainly nursing assistants). Responders were predominantly female (69.1%), Indo-Trinidadian (55.8%), Christian (47.5%), self-claimed "very religious" (48.3%), and had <5 years of working experience (40.6%). The prevalence of CAM use was 92.4% for nurses, 64.9% for doctors, 83.3% for pharmacists, and 77.1% for other health care providers. The majority (50-75%) reported fair knowledge of herbal, spiritual, alternative, and physical types of CAM, but had no knowledge of energy therapy and therapeutic methods. Sex, ethnicity, and type of health care provider were associated with both personal use and recommendation for the use of CAM. Predictors of CAM use were sex, religion, and type of health care provider; predictors of recommendation for the use of CAM were sex and type of health care provider. About half of health care providers (51.4%) and doctors (52%) were likely to ask their patients about CAM and <15% were likely to refer patients to a CAM practitioner. However, health care providers expressed interest in being educated on the subject. Doctors (51.9%) and pharmacists (63.3%) said that combination therapy is superior to conventional medicine alone. Less than 10% said conventional medicine should be used alone. Knowledge about CAM is low among health care providers. The majority engages in using CAM but is reluctant to recommend it. Predictors of CAM use were sex, religion, and profession; predictors of recommendation for the use of CAM were sex and profession. Health care providers feel the future lies in integrative medicine.


Divergent Molecular Phenotypes in Point Mutations at the Same Residue in Beta-Myosin Heavy Chain Lead to Distinct Cardiomyopathies.
In genetic cardiomyopathies, a frequently described phenomenon is how similar mutations in one protein can lead to discrete clinical phenotypes. One example is illustrated by two mutations in beta myosin heavy chain (β-MHC) that are linked to hypertrophic cardiomyopathy (HCM) (Ile467Val, I467V) and left ventricular non-compaction (LVNC) (Ile467Thr, I467T). To investigate how these missense mutations lead to independent diseases, we studied the molecular effects of each mutation using recombinant human β-MHC Subfragment 1 (S1) in in vitro assays. Both HCM-I467V and LVNC-I467T S1 mutations exhibited similar mechanochemical function, including unchanged ATPase and enhanced actin velocity but had opposing effects on the super-relaxed (SRX) state of myosin. HCM-I467V S1 showed a small reduction in the SRX state, shifting myosin to a more actin-available state that may lead to the "gain-of-function" phenotype commonly described in HCM. In contrast, LVNC-I467T significantly increased the population of myosin in the ultra-slow SRX state. Interestingly, molecular dynamics simulations reveal that I467T allosterically disrupts interactions between ADP and the nucleotide-binding pocket, which may result in an increased ADP release rate. This predicted change in ADP release rate may define the enhanced actin velocity measured in LVNC-I467T, but also describe the uncoupled mechanochemical function for this mutation where the enhanced ADP release rate may be sufficient to offset the increased SRX population of myosin. These contrasting molecular effects may lead to contractile dysregulation that initiates LVNC-associated signaling pathways that progress the phenotype. Together, analysis of these mutations provides evidence that phenotypic complexity originates at the molecular level and is critical to understanding disease progression and developing therapies.


Detection and identification of transient intermediates in the reactions of tryptophan synthase with oxindolyl-L-alanine and 2,3-dihydro-L-tryptophan. Evidence for a tetrahedral (gem-diamine) intermediate.
The reactions of 2,3-dihydro-L-tryptophan (DHT) and oxindolyl-L-alanine (OXA) with tryptophan synthase have been investigated by rapid-scanning stopped-flow (RSSF) spectroscopy and by the concentration dependence of rates measured by single-wavelength stopped-flow (SWSF) spectroscopy. The RSSF spectral changes for DHT and OXA show the disappearance of the internal aldimine (lambda max 412 nm), the formation and decay of intermediates absorbing less than or equal to 340 nm, and the appearance of the quinonoid (lambda max 492 and 480 nm, respectively). Rate constants determined by SWSF were either well resolved (i.e., k1[DHT], k-1 greater than k2, k-2 greater than k3, k-3) or indicative of a tightly coupled system (i.e., k1[OXA], k-1 greater than or equal to k2, k-2 greater than k3, k-3). The RSSF spectral changes and SWSF kinetic studies together with computer simulations of the kinetic time courses are consistent with a mechanism that includes formation of a bleached species. Detection of these shorter wavelength species in the reactions of OXA and DHT indicates that substrate analogues with tetrahedral geometry at C-3 induce new protein-substrate interactions that result in the accumulation of species not previously detected in the tryptophan synthase system. The bleached species with lambda max less than or equal to 340 nm are proposed as the gem-diamine intermediates.


Algorithm for neuropathic pain treatment: an evidence based proposal.
New studies of the treatment of neuropathic pain have increased the need for an updated review of randomized, double-blind, placebo-controlled trials to support an evidence based algorithm to treat neuropathic pain conditions. Available studies were identified using a MEDLINE and EMBASE search. One hundred and five studies were included. Numbers needed to treat (NNT) and numbers needed to harm (NNH) were used to compare efficacy and safety of the treatments in different neuropathic pain syndromes. The quality of each trial was assessed. Tricyclic antidepressants and the anticonvulsants gabapentin and pregabalin were the most frequently studied drug classes. In peripheral neuropathic pain, the lowest NNT was for tricyclic antidepressants, followed by opioids and the anticonvulsants gabapentin and pregabalin. For central neuropathic pain there is limited data. NNT and NNH are currently the best way to assess relative efficacy and safety, but the need for dichotomous data, which may have to be estimated retrospectively for old trials, and the methodological complexity of pooling data from small cross-over and large parallel group trials, remain as limitations.


Combined fine-needle aspiration with core needle biopsy for assessing thyroid nodules: a more valuable diagnostic method?
This study aimed to evaluate the diagnostic value of combined fine-needle aspiration (FNA) with core needle biopsy (CNB) in thyroid nodules. FNA and CNB were performed simultaneously on 703 nodules. We compared the proportions of inconclusive results and the diagnostic performance for malignancy among FNA, CNB, and combined FNA/CNB for different nodule sizes. Combined FNA/CNB showed lower proportions of inconclusive results than CNB for all nodules (2.8% vs. 5.7%, P<0.001), nodules ≤1.0 cm (4.9% vs. 7.3%, P=0.063), nodules >1.0 cm (2.0% vs. 5.0 %, P<0.001), nodules ≤1.5 cm (3.8% vs. 7.9 %, P<0.001), and nodules >1.5 cm (2.1% vs. 3.9 %, P=0.016). The sensitivity of combined FNA/CNB in predicting malignancy was significantly higher than that of CNB (89.0% vs. 80.0%, P<0.001) and FNA (89.0% vs. 58.1%, P<0.001) for all nodules. Within American College of Radiology Thyroid and Imaging Reporting and Data System grades 4-5, in the subgroup of nodules ≤1.5 cm, combined FNA/ CNB showed the best sensitivity in predicting malignancy (91.4%), significantly higher than that of CNB (81.0%, P<0.001) and FNA (57.8%, P<0.001). However, in the subgroup of nodules >1.5 cm, the difference between combined FNA/CNB and CNB was not significant (84.2% vs. 78.9%, P=0.500). Regardless of nodule size, combined FNA/CNB tended to yield lower proportions of inconclusive results than CNB or FNA alone and exhibited higher performance in diagnosing malignancy. The combined FNA/CNB technique may be a more valuable diagnostic method for nodules ≤1.5 cm and nodules with a risk of malignancy than CNB and FNA alone.


Chlamydia trachomatis immune evasion via downregulation of MHC class I surface expression involves direct and indirect mechanisms.
Genital C. trachomatis infections typically last for many months in women. This has been attributed to several strategies by which C. trachomatis evades immune detection, including well-described methods by which C. trachomatis decreases the cell surface expression of the antigen presenting molecules major histocompatibility complex (MHC) class I, MHC class II, and CD1d in infected genital epithelial cells. We have harnessed new methods that allow for separate evaluation of infected and uninfected cells within a mixed population of chlamydia-infected endocervical epithelial cells to demonstrate that MHC class I downregulation in the presence of C. trachomatis is mediated by direct and indirect (soluble) factors. Such indirect mechanisms may aid in priming surrounding cells for more rapid immune evasion upon pathogen entry and help promote unfettered spread of C. trachomatis genital infections.


Oscillatory cerebral blood flow is associated with impaired neurocognition and functional hyperemia in postural tachycardia syndrome during graded tilt.
We hypothesize that upright cognitive impairment in patients with postural tachycardia syndrome (POTS) is caused by reduced cerebral blood flow (CBF). The CBF velocity (CBF(v)) measured by transcranial Doppler ultrasound decreased excessively during 70° tilt in a minority of patients with intermittent hyperpnea/hypocapnia. Incremental tilt showed no difference in mean CBF(v). But N-back memory tasking indicated progressive compromised memory, reduced functional hyperemia, and reduced neurovascular coupling. Orthostasis caused slow oscillations in CBF(v) linked to oscillations in arterial pressure in patients with POTS. We also hypothesize that oscillatory CBF(v) degrades neurovascular coupling. We performed 2-back testing when subjects were in supine position and during incremental tilts to 15°, 30°, 45°, and 60° in 11 patients with POTS and 9 controls. Oscillatory arterial pressure, oscillatory CBF(v), and neurovascular coupling were similar in supine position. The oscillatory arterial pressure increased by 31%, 45%, 67%, and 93% in patients with POTS during tilt and remained unchanged in the controls. Oscillatory CBF(v) increased by 61%, 82%, 161%, and 264% in patients with POTS during tilt and remained unchanged in the controls. Functional hyperemia decreased from 4.1% to 3.0%, 1.1%, 0.2%, and to 0.04% in patients with POTS, but it was unchanged at 4% in the controls. Percent correct N-back responses decreased from 78% to 33% in patients with POTS, whereas they remained at 89% in the controls. In patients with POTS, oscillatory CBF(v) was linearly correlated with functional hyperemia (r(2)=0.76). Increased oscillatory CBF is associated with reduced neurovascular coupling and diminished cognitive performance in patients with POTS.


Modern Patellofemoral Arthroplasty.
The ideal procedure for isolated patellofemoral arthritis is a controversial topic. Patellofemoral arthroplasty (PFA) is an option that aims to restore normal kinematics to the knee while preserving bone. PFA has been shown to have benefits compared with total knee arthroplasty (TKA) in this patient population but has historically had a high failure rate. Revision rates are improving with modern implants and tight indications but still remain higher than TKA. This review summarizes current thinking around PFA using modern implants and techniques in 2023, provides an implant-specific analysis, and assesses how we can improve outcomes after PFA based on the current literature. The aim was to provide an outline of the evidence around PFA on which surgeons can make decisions to optimize patient outcome in this young and active population. Four databases (MEDLINE, Embase, Scopus, and SPORTDiscus) were searched for concepts of patellofemoral joint arthroplasty. After abstract and text review, a screening software was used to assess articles based on inclusion criteria for studies describing indications, outcomes, and techniques for isolated PFA using modern implants, with or without concomitant procedures. A total of 191 articles were included for further examination, with 62 articles being instructional course lectures, systematic reviews, technique articles, narrative reviews, expert opinions, or meta-analyses. The remaining articles were case reports, trials, or cohort studies. Articles were used to create a thorough outline of multiple recurrent topics in the literature. PFA is an appealing option that has the potential to provide a more natural feeling and functioning knee for those with isolated PF arthritis. The high rate of revision is a cause for concern and there are several technical details that should be stressed to optimize results. The uncertain outcome after revision to TKA also requires more investigation. In addition, the importance of strict selection criteria and firm indications cannot be stressed enough to optimize longevity and attempt to predict those who are likely to have progression of tibiofemoral osteoarthritis. The development of new third-generation implants is promising with excellent functional outcomes and a much lower rate of maltracking and implant complications compared with earlier generations. The impact of these implants and improvement in surgical techniques on the revision rate of PFA will be determined from longer-term outcomes.


Effect of Tooth-Colored Restorative Materials on Reliability of Heat-Pressed Lithium Disilicate.
Restorative material selection in complete mouth rehabilitation is an important factor in long-term management of potential technical complications. The aim of this study was to evaluate in vitro the reliability (fracture resistance) of lithium disilicate fatigued with different restorative materials. A step-stress accelerated life-testing model was used. Seventy disc specimens were heat-pressed. Five groups of different indenter materials fatigued the lithium-disilicate specimens: group WC (tungsten carbide served as a control), group PR (interpenetrating polymer network [IPN] resin-based denture tooth), group POM (heat-pressed leucite glass-ceramic), group LD (heat-pressed lithium disilicate), and group ZR (zirconium dioxide). Lithium-disilicate specimens were randomly divided into four groups (n = 14). Specimens were fatigued to failure according to three step-stress profiles: light, moderate, and aggressive. Use level probability Weibull plots were generated, and each group's reliability, failure rate, and mean life to failure were calculated. The IPN resin-based denture tooth group had the highest reliability and mean life to failure, and lowest failure rate as compared to lithium disilicate and zirconium dioxide. No significant difference existed between the reliability of the tungsten carbide and leucite glass-ceramic groups and the IPN resin-based denture tooth group. Lithium-disilicate specimens fatigued with IPN resin-based denture teeth exhibited higher reliability than specimens fatigued with lithium disilicate and zirconium dioxide. There was a difference in fracture characteristics in lithium-disilicate specimens fatigued with tungsten carbide, lithium disilicate, and zirconium dioxide, versus those fatigued with IPN resin-based denture teeth and leucite glass-ceramic material.


A large randomized individual and group intervention conducted by registered dietitians increased adherence to Mediterranean-type diets: the PREDIMED study.
To assess the effectiveness of an intervention aimed to increase adherence to a Mediterranean diet. A 12-month assessment of a randomized primary prevention trial. One thousand five hundred fifty-one asymptomatic persons aged 55 to 80 years, with diabetes or > or =3 cardiovascular risk factors. Participants were randomly assigned to a control group or two Mediterranean diet groups. Those allocated to the two Mediterranean diet groups received individual motivational interviews every 3 months to negotiate nutrition goals, and group educational sessions on a quarterly basis. One Mediterranean diet group received free virgin olive oil (1 L/week), the other received free mixed nuts (30 g/day). Participants in the control group received verbal instructions and a leaflet recommending the National Cholesterol Education Program Adult Treatment Panel III dietary guidelines. Changes in food and nutrient intake after 12 months. Paired t tests (for within-group changes) and analysis of variance (for between-group changes) were conducted. Participants allocated to both Mediterranean diets increased their intake of virgin olive oil, nuts, vegetables, legumes, and fruits (P<0.05 for all within- and between-group differences). Participants in all three groups decreased their intake of meat and pastries, cakes, and sweets (P<0.05 for all). Fiber, monounsaturated fatty acid, and polyunsaturated fatty acid intake increased in the Mediterranean diet groups (P<0.005 for all). Favorable, although nonsignificant, changes in intake of other nutrients occurred only in the Mediterranean diet groups. A 12-month behavioral intervention promoting the Mediterranean diet can favorably modify an individual's overall food pattern. The individual motivational interventions together with the group sessions and the free provision of high-fat and palatable key foods customary to the Mediterranean diet were effective in improving the dietary habits of participants in this trial.


Identifying clusters of coexisting conditions and outcomes among adults admitted to hospital with community-acquired pneumonia: a multicentre cohort study.
Little is known about patterns of coexisting conditions and their influence on clinical care or outcomes in adults admitted to hospital for community-acquired pneumonia (CAP). We sought to evaluate how coexisting conditions cluster in this population to advance understanding of how multimorbidity affects CAP. We studied 11 085 adults admitted to hospital with CAP at 7 hospitals in Ontario, Canada. Using cluster analysis, we identified patient subgroups based on clustering of comorbidities in the Charlson Comorbidity Index. We derived and replicated cluster analyses in independent cohorts (derivation sample 2010-2015, replication sample 2015-2017), then combined these into a total cohort for final cluster analyses. We described differences in medications, imaging and outcomes. Patients clustered into 7 subgroups. The low comorbidity subgroup (n = 3052, 27.5%) had no comorbidities. The DM-HF-Pulm subgroup had prevalent diabetes, heart failure and chronic lung disease (n = 1710, 15.4%). One disease category defined each remaining subgroup, as follows: pulmonary (n = 1621, 14.6%), diabetes (n = 1281, 11.6%), heart failure (n = 1370, 12.4%), dementia (n = 1038, 9.4%) and cancer (n = 1013, 9.1%). Corticosteroid use ranged from 11.5% to 64.9% in the dementia and pulmonary subgroups, respectively. Piperacillin-tazobactam use ranged from 9.1% to 28.0% in the pulmonary and cancer subgroups, respectively. The use of thoracic computed tomography ranged from 5.7% to 36.3% in the dementia and cancer subgroups, respectively. Adjusting for patient factors, the risk of in-hospital death was greater in the cancer (adjusted odds ratio [OR] 3.12, 95% confidence interval [CI] 2.44-3.99), dementia (adjusted OR 1.57, 95% CI 1.05-2.35), heart failure (adjusted OR 1.66, 95% CI 1.35-2.03) and DM-HF-Pulm subgroups (adjusted OR 1.35, 95% CI 1.12-1.61), and lower in the diabetes subgroup (adjusted OR 0.67, 95% CI 0.50-0.89), compared with the low comorbidity group. Patients admitted to hospital with CAP cluster into clinically recognizable subgroups based on coexisting conditions. Clinical care and outcomes vary among these subgroups with little evidence to guide decision-making, highlighting opportunities for research to personalize care.


Biodegradable polyphosphoester micelles for gene delivery.
A new biodegradable polyphosphoester, poly[[(cholesteryl oxocarbonylamido ethyl) methyl bis(ethylene) ammonium iodide] ethyl phosphate] (PCEP) was synthesized and investigated for gene delivery. Carrying a positive charge in its backbone and a lipophilic cholesterol structure in the side chain, PCEP self-assembled into micelles in aqueous buffer at room temperature with an average size of 60-100 nm. It could bind and protect plasmid DNA from nuclease digestion. Cell proliferation assay indicated a lower cytotoxicity for PCEP than for poly-L-lysine and Lipofectamine. The IC50 determined by the WST-1 assay was 69.8, 51.6, and 12.1 microg/mL for PCEP, Lipofectamine, and poly-L-lysine, respectively. PCEP efficiently delivered DNA to several cell lines such as HEK293, Caco-2, and HeLa. The highest efficiency was achieved when PCEP/DNA complex was prepared in Opti-MEM with a +/- charge ratio of 1.5-2. The transfection efficiency did not change significantly when the complex was used 3 days after preparation. The addition of chloroquine to the formulation increased transfection efficiency 10- to 50-fold compared to the complex alone. In vivo studies showed a luciferase expression by PCEP/DNA complexes in muscle increasing with time during 3 months, although the expression level was lower than that by direct injection of naked DNA. In addition to biodegradability and lower toxicity, the PCEP micelle carrier offers structural versatility. The backbone charge density and the side chain lipophilicity are two parameters that can be varied through copolymerization and monomer variation to optimize the transfection efficiency.


The joint effects of water and sanitation on diarrhoeal disease: a multicountry analysis of the Demographic and Health Surveys.
To assess whether the joint effects of water and sanitation infrastructure, are acting antagonistically (redundant services preventing the same cases of diarrhoeal disease), independently, or synergistically; and to assess how these effects vary by country and over time. We used data from 217 Demographic and Health Surveys conducted in 74 countries between 1986 and 2013. We used modified Poisson regression to assess the impact of water and sanitation infrastructure on the prevalence of diarrhoea among children under 5. The impact of water and sanitation varied across surveys, and adjusting for socio-economic status drove these estimates towards the null. Sanitation had a greater effect than water infrastructure when all 217 surveys were pooled; however, the impact of sanitation diminished over time. Based on survey data from the past 10 years, we saw no evidence for benefits in improving drinking water or sanitation alone, but we estimated a 6% reduction of both combined (prevalence ratio = 0.94, 95% confidence limit 0.91-0.98). Water and sanitation interventions should be combined to maximise the number of cases of diarrhoeal disease prevented in children under 5. Further research should identify the sources of variability seen between countries and across time. These national surveys likely include substantial measurement error in the categorisation of water and sanitation, making it difficult to interpret the roles of other pathways.


Management and outcome of complex hypospadias repairs.
Repeated attempts at surgical repair of hypospadias may leave the penis scarred, hypovascular and shortened. We report on the clinical outcomes of our repaired complex hypospadias cases, often referred to as hypospadias cripples. We evaluated the records of 137 children and young adults from January 1980 through December 2002 who were referred to us after multiple unsuccessful hypospadias repairs. The records of 11 patients were inadequate. The ages of the remaining 126 patients ranged from 14 months to 35 years. The number of prior surgical procedures ranged from 2 to 23. Of the 126 cases 98 (78%) were repaired in a single stage (group 1) and 28 (22%) underwent multistage repairs (group 2). Major complications occurred in 17 cases (17%) in group 1 and in 2 (7%) in group 2. Major complications included repair breakdown, stricture, diverticulum and multiple fistulas. Minor complications occurred in 9 (9%) cases in group 1 and in 4 (14%) in group 2. Minor complications included a single small urethrocutaneous fistula, skin tethering, inclusion cysts and glandular irregularity. Urethral substitution using skin grafts and/or bladder mucosa resulted in a high complication rate of 32% and 37%, respectively, whereas use of buccal mucosa resulted in a 15% complication rate. Preoperative psychological counseling and discussion with other parents or patients were helpful. We no longer use free skin grafts and/or bladder mucosa for urethral substitution. Waterproofing the urethral tube was performed using either a dartos or tunica vaginalis flap. When the quality of the tissues was poor or severely scarred a 2-stage repair was performed. When resurfacing the penis local skin flaps may be insufficient or unreliable, and rotation of scrotal skin flaps or burial of the penis in the scrotum (Cecil-Culp) offers dependable skin coverage.


Ecophysiology and Growth of White Spruce Seedlings from Various Seed Sources along a Climatic Gradient Support the Need for Assisted Migration.
With climate change, favorable growing conditions for tree species are shifting northwards and to higher altitudes. Therefore, local populations are becoming less adapted to their environment. Assisted migration is one of the proposed adaptive measures to reduce the vulnerability of natural populations and maintain forest productivity. It consists of moving genetic material to a territory where future climate conditions correspond to those of its current location. Eight white spruce (Picea glauca [Moench] Voss) seed sources representing as many seed orchards were planted in 2013 at three forest sites simulating a south-north climatic gradient of 1.7°C in Québec, Canada. The objectives were to (1) evaluate the morpho-physiological responses of the different seed sources and (2) determine the role of genetic adaptation and physiological plasticity on the observed variation in morpho-physiological traits. Various seedling characteristics were measured, notably height growth from nursery to the fourth year on plantation. Other traits such as biomass and carbon allocation, nutritional status, and various photosynthetic traits before bud break, were evaluated during the fourth growing season. No interaction between sites and seed sources was observed for any traits, suggesting similar plasticity between seed sources. There was no change in the rank of seed sources and sites between years for height growth. Moreover, a significant positive correlation was observed between the height from the nursery and that after 4 years in the plantation. Southern seed sources showed the best height growth, while optimum growth was observed at the central site. Juvenile height growth seems to be a good indicator of the juvenile carbon sequestration and could serve as a selection criterion for the best genetics sources for carbon sequestration. Vector analysis showed no nitrogen deficiency 4 years after planting. Neither seed sources nor planting sites had a significant effect on photosynthesis before bud break. The observed results during the establishment phase under different site conditions indicate that southern seed sources may already benefit from assisted migration to cooler climatic conditions further north. While northern seed sources are likely to benefit from anticipated local global warming, they would not match the growth performance of seedlings from southern sources.


[Respiratory effects of antihypertensive agents acting on alpha adrenergic receptors].
There are several factors involved in the bronchial muscle tone regulation, such as biochemical, nervous and humoral mechanisms. In general, in chronic obstructive bronchopneumopathy there is a reduction in air flow, due to different physiopathologic factors which, alone or combined, produce these disorders. In particular, according to suggestive hypotheses, it seems that asthma is due to a partial or complete block of beta-receptors with a prevalence of alpha-receptors and subsequent bronchial hypertonicity and hyperactivity, while in patients with allergic asthma a deficiency of beta-2-adrenergic system and an hyperresponsiveness of alpha-adrenergic and/or cholinergic system have been suggested. In relation to this part, alpha-1 antagonist drugs demonstrated to have a bronchodilator effect, thanks to their mechanism of action. Examples of this class of drugs are prazosin, indoramin, and an analogous of prazosin, doxazosin, which has a high antagonist and selective action for post-synaptic alpha-1 adrenergic receptors and a longer half-life. Several and recent observations show that these drugs do not interfere with the respiratory function principal parameters, both in healthy and in asthmatic or chronic bronchopathy subjects, but produce an appreciable improvement of these parameters. Presynaptic alpha-2-adrenergic receptors agonist drugs would mainly act by inhibiting the central nervous system, with a subsequent lower stimulation of post-synaptic alpha-1-adrenergic receptors. Examples of this class of drugs are clonidine, rilmenidine and guanabenz (but, in this case, the effects on the respiratory function are more controversial).(ABSTRACT TRUNCATED AT 250 WORDS)