[Evaluation of effectiveness of the micrographia's therapy in Parkinson's disease patients].
Micrographia is one of the characteristic symptoms of Parkinson's disease. Its cause is hypokinesia (reduced movement's amplitude). The methods of management of micrographia are founded on the assumption that the important cause of prolonged hypokinesia in Parkinson's disease patients is impairment of attention. The aim of the work was to analyse relationship between an occurrence and intensity of micrographia and others clinical features of Parkinson's disease. The another aim was to assessment of effectiveness of micrographia's management. 40 Parkinson's disease patients (19 women and 21 men) aged from 49 to 82 years (mean--65.5) were examined. In the analysis age, duration of disease, predominance of right- or left - side symptoms, impairments of concentration of attention, dosage of levodopa, UPDRS score, and the level on the Hoehn and Yahr's scale were taken into account. In therapy of micrographia three methods were managed: writing between lines determining desired height of letters, writing with a big dot as a guide for letter's size and regular reminding the patient of the need to write at bigger size. micrographia was occurred in 30 of 40 Parkinson's disease patients (75%). The better writing magnitude was obtained in all patients, but in 16 patients the effect was much better and maintained to 6 hours, whereas in 14 patients the effect was worse and maintained only short time. No relationship was found between an applied method of therapy and its effect. A significant statistical relationship was established between impairments of concentration of attention and a worse effect micrographia's therapy. Micrographia occurs in 75% Parkinson's disease patients. The therapy is successful in more than 50% of patients. Impairments of concentration of attention exert an adverse effect on micrographia's therapy.


Association between Vitamin D Deficiency and Clinical Parameters in Men and Women Aged 50 Years or Older: A Cross-Sectional Cohort Study.
Vitamin D deficiency (VDD) is increasingly prevalent on a global scale and is connected to chronic health issues including diabetes, obesity, and inflammation. This study aimed to investigate the association between VDD and various clinical parameters including glycated hemoglobin (HbA1c), body mass index (BMI), and inflammatory markers. This cross-sectional cohort study included Korean men and women aged 50 years and older (290 men, 125 women); VDD was classified as serum 25-hydroxyvitamin D (25[OH]D) levels below 20 ng/mL. Vitamin D deficiency was more prevalent in men (64.5%) compared to that in women (35.2%). Men with VDD had higher fat mass and HbA1c levels, lower muscle strength, and worse physical performance. Among women, VDD was associated with higher BMI, HbA1c, tumor necrosis factor-alpha (TNF-α), and creatinine levels. In women, 25(OH)D levels exhibited an inverse relationship with HbA1c, BMI, and TNF-α concentrations. However, there were no differences in the levels of interleukin-6 and interleukin-1 beta according to vitamin D status in both men and women. Vitamin D deficiency is linked to higher HbA1c, BMI, and inflammatory markers in older Korean women, thus warranting the maintenance of sufficient vitamin D levels for overall health.


The Impact of the Community Psychiatric Treatment Program on the User's Quality of Life - experience of the Study Participants.
Community psychiatric treatment (CPT) is a novelty in the primary health care system for the people with mental health problems. The service operates according to the Flexible Assertive Community Treatment (FACT) model. FACT is a rehabilitation-oriented clinical case management model based on the ACT model (Assertive Community Treatment). This study aimed to assess how involvement in a community psychiatry program affects the quality of life of program participants. Between December 2020 and March 2021 we conducted a qualitative study. To obtain information, semi-structured interviews with twelve patients treated in the Community Psychiatric Treatment program were conducted. All restriction regarding COVID pandemic were considered at the research. Content analysis of the text was used for data analysis, which means that we interpreted the analysed text and determine the meaning of individual parts of the text. Twelve persons participated in the interviews, six of which were women (50%) and six men (50%). All the participants were of Slovenian nationality. They were from 28 to 80 years old, with a mean age of 47 years. We identified twelve categories and four themes: treatment problems, principles of operating of CPT program, satisfaction with inclusion in the CPT and improvement of quality of life through inclusion in the program. Quality of life is essential for every individual, as well as for people with mental health problems. In our study we have proved that the Community psychiatric programs could significantly impact the quality of life of people with mental health problems.


Stress regulation in adolescents: physiological reactivity during the adult attachment interview and conflict interaction.
The current study examined whether adolescents' attachment representations were associated with differences in emotion regulation during the Adult Attachment Interview (AAI; C. George, N. Kaplan, & M. Main, 1996) and during a mother-adolescent conflict interaction task (Family Interaction Task [FIT]; J. P. Allen et al., 2003). Participants were one hundred and fifty-six 14-year-old adolescents. Dismissing adolescents showed less interbeat interval (IBI) reactivity (indicating less stress) during the AAI than secure adolescents. However, during the FIT, dismissing adolescents showed more IBI reactivity. No differences in physiological reactivity were found between individuals with resolved or unresolved loss or trauma during the AAI or FIT. The results indicate that dismissing adolescents may effectively use a defensive strategy during the AAI but less so in direct conflict interaction with their attachment figure.


[Preparation of controlled release microsphere incorporating bFGF and its effect on Schwann cells].
To study the preparation method of bFGF microspheres and to investigate the bioactivities of bFGF, which were released from the bFGF microspheres, on the cultured schwann cells. bFGF was microcapsulated with the multiple emulsion encapsulative method using PLGA as coating material. Its morphology, particle size distribution, drug loading-embedding rate and in vitro release property were studied. The cultured schwann cells were grouped according to the different ingredients being added to the culture medium: bFGF group, bFGF-PLGA group. Then the number, the viability and the cell cycle of schwann cells were measured. The morphology and the particle size distribution of the bFGF-PLGA microspheres were even and good; the drug-loading and drug-embedding rate of microspheres were (27.18 x 10(-3)) % +/- (0.51 x 10(-3)) %, 66. 43% +/- 1.24%; the release property of microspheres in vitro was good and the overall release rate was 72. 47% in 11 days. The in vitro cellular study showed: 1, 2 days after plate culture, the cell number and cell viability of bFGF group was much better than that of bFGF-PLGA group; 3, 4 days after plate culture, the cell number and cell viability of bFGF group and bFGF-PLGA group were not different statistically; 6, 8 days after plate culture, the cell number and cell viability of bFGF-PLGA group was much better than that of bFGF group. Through the flow cytometry examination: 2 days after plate culture, the GJ/M+S percentage of bFGF group was higher than that of bFGF-PLGA group; 4, 8 days after plate culture, the G2/M+S percentage of bFGF-PLGA group was higher than that of bFGF group. So, it is practical to prepare the bFGF-PLGA microspheres with the multiple emulsion encapsulative method. bFGF-PLGA microspheres can preserve the bioactivities of bFGF effectively and promotes the proliferation of schwann cells in a long period because of the controlled release of bFGF from microspheres.


Analysis of the miRNA profile in C6/36 cells persistently infected with dengue virus type 2.
Dengue virus (DENV) is the most important arbovirus in the world; DENV is transmitted by the Aedes genus of mosquitoes and can establish a life-long persistent infection in mosquitoes. However, the exact mechanism by which persistent infection is established remains unknown. In this study the differential expression of miRNAs was analysed by deep sequencing and RT-qPCR using a previously established C6/36-HT cell line persistently infected with DENV 2 (C6-L) as a model. miR-927, miR-87, miR-210, miR-2a-3p, miR-190 and miR-970 were up-regulated, whereas miR-252, miR-263a-3p, miR-92b, miR-10-5p miR-9a-5p, miR-9a-1, miR-124, miR-286a and miR-286b were down-regulated in C6-L cells compared with C6/36 cells acutely infected with the same virus or mock-infected cells. Deep sequencing results were validated by RT-qPCR for the highly differentially expressed miR-927 and miR-9a-5p, which were up- and down-regulated, respectively, compared with both acutely and mock-infected C6/36 cells. The putative targets of these miRNAs include components of the ubiquitin conjugation pathway, vesicle-mediated transport, autophagy, and the JAK-STAT cascade as well as proteins with endopeptidase activity. Other putative targets include members of the Toll signalling pathway and proteins with kinase, ATPase, protease, scavenger receptor or Lectin C-type activity or that participate in fatty acid biosynthesis or oxidative stress. Our results suggest that several specific miRNAs help regulate the cellular functions that maintain equilibrium between viral replication and the antiviral response during persistent infection of mosquito cells. This study is the first report of a global miRNA profile in a mosquito cell line persistently infected with DENV.


Interferon-alpha regulation of lymphocyte function in systemic lupus erythematosus.
Interferon (IFN) production and responsiveness are abnormal in some patients with systemic lupus erythematosus (SLE). The present investigation was designed to further delineate these abnormalities of IFN response. The response to IFN-alpha was determined in peripheral blood mononuclear cells (PBMC) using the natural killer (NK) cell assay, the concanavalin A (Con A)-induced T-cell proliferation assay, and the pokeweed mitogen (PWM) blastogensis system. The average NK cell activity was impaired in SLE patients (19.7 +/- 3.8 LU/10(7) cells; n = 22) compared to 24 normal controls (45.6 +/- 7.2 LU/10(7) cells; n = 24) (P less than 0.05). In addition, the response to IFN in the NK cell system was impaired in SLE (110.0 +/- 38.0% enhancement) compared to controls (320.0 +/- 94.0% enhancement; +/- SD P less than 0.05). In contrast to the impaired IFN response of the NK cell in SLE, the effect of IFN on Con A- and PWM-induced blastogenesis in SLE was normal (P greater than 0.1). Thus, the defect in IFN response in SLE appeared to be confined to the NK cell system, but was not present in the other assay systems. To determine if the impaired IFN response might be secondary to impaired release of natural killer cytotoxic factor (NKCF) induced by IFN, NKCF was generated from PBMC in the presence of IFN. IFN-induced NKCF release was markedly impaired in SLE (8.9 +/- 11.7%) relative to normal controls (22.3 +/- 11.2%) (P less than 0.05). IFN-induced release of NKCF in SLE also closely correlated with IFN-induced NK enhancement (r = .83, P less than 0.05). No depletion of NK cells was noted using the monoclonal antibody HNK-1. Thus, this study demonstrates for the first time that the insensitivity to IFN in SLE is not a universal lymphocyte defect but appears to be isolated to the NK cell which is functionally abnormal.


[Analysis of certain nodal features of regenerative reactions].
The increased degree of death of cells due to increased functional stresses may serve as a starting factor responsible for the compensatory growth. This fact allows considering reparative regeneration and compensatory growth as phenomena of the same class. Different forms of reparative growth (intracellular regeneration and proliferation of cells, endomorphosis and epimorphosis) serve to concrete tasks of reparation at different levels of organization of the morphological substrate. They are sustained by endogeneous morphogenetical mechanisms developing independently of other levels and of environment. In the subsystem of lover levels of organization the involved morphogenetical mechanisms may sustain not only compensation of the lost structures but also their overproduction. As a rule the program of reparative processes may be realized on higher (e. g. organic) level but partly. Among available criteria of regeneration at the present time the calculation of mitoses should be considered the most adequate one. The systemic approach may prove to be a perspective method both for estimation of regeneratory effect and for deeper analysis of reparative phenomena in general.


Evaluation of Adjusted and Unadjusted Indirect Comparison Methods in Benefit Assessment. A Simulation Study for Time-to-event Endpoints.
With the Act on the Reform of the Market for Medicinal Products (AMNOG) in Germany, pharmaceutical manufacturers are obliged to submit a dossier demonstrating added benefit of a new drug compared to an appropriate comparator. Underlying evidence was planned for registration purposes and therefore often does not meet the appropriate comparator as defined by the Federal Joint Committee (G-BA). For this reason AMNOG allows indirect comparisons to assess the extent of added benefit. The aim of this study is to evaluate the characteristics and applicability of adjusted indirect comparison described by Bucher and Matching-Adjusted Indirect Comparison (MAIC) in various situations within the early benefit assessment according to §35a Social Code Book 5. In particular, we consider time-to-event endpoints. We conduct a simulation study where we consider three different scenarios: I) similar study populations, II) dissimilar study populations without interactions and III) dissimilar study populations with interactions between treatment effect and effect modifiers. We simulate data from a Cox model with Weibull distributed survival times. Desired are unbiased effect estimates. We compare the power and the proportion of type 1 errors of the methods. I) Bucher and MAIC perform equivalently well and yield unbiased effect estimates as well as proportions of type 1 errors below the significance level of 5 %. II) Both Bucher and MAIC yield unbiased effect estimates, but Bucher shows a higher power for detection of true added benefit than MAIC. III) Only MAIC, but not Bucher yields unbiased effect estimates. When using robust variance estimation MAIC yields a proportion of type 1 error close to 5 %. In general, power of all methods for indirect comparisons is low. An increasing loss of power for the indirect comparisons can be observed as the true treatment effects decrease. Due to the great loss of power and the potential bias for indirect comparisons, head-to-head trials using the appropriate comparator as defined by the Federal Joint Committee should be conducted whenever possible. However, indirect comparisons are needed if no such direct evidence is available. To conduct indirect comparisons in case of a present common comparator and similar study populations in the trials to be compared, both Bucher and MAIC can be recommended. In case of using adjusted effect measures (such as Hazard Ratio), the violation of the similarity assumption has no relevant effect on the Bucher approach as long as interactions between treatment effect and effect modifiers are absent. Therefore Bucher can still be considered appropriate in this specific situation. In the authors' opinion, MAIC can be considered as an option (at least as sensitivity analysis to Bucher) if such interactions are present or cannot be ruled out. Nevertheless, in practice MAIC is potentially biased and should always be considered with utmost care.


Isolation and analysis of differentially expressed genes in Penicillium glabrum subjected to thermal stress.
Penicillium glabrum is a filamentous fungus frequently involved in food contamination. Numerous environmental factors (temperature, humidity, atmospheric composition, etc.) or food characteristics (water activity, pH, preservatives, etc.) could represent potential sources of stress for micro-organisms. These factors can directly affect the physiology of these spoilage micro-organisms: growth, conidiation, synthesis of secondary metabolites, etc. This study investigated the transcriptional response to temperature in P. glabrum, since this factor is one of the most important for fungal growth. Gene expression was first analysed by using suppression subtractive hybridization to generate two libraries containing 445 different up- and downregulated expressed sequence tags (ESTs). Expression of these ESTs was then assessed for different thermal stress conditions, with cDNA microarrays, resulting in the identification of 35 and 49 significantly up- and downregulated ESTs, respectively. These ESTs encode heat-shock proteins, ribosomal proteins, superoxide dismutase, trehalose-6-phosphate synthase and a large variety of identified or unknown proteins. Some of these may be molecular markers for thermal stress response in P. glabrum. To our knowledge, this work represents the first study of the transcriptional response of a food spoilage filamentous fungus under thermal stress conditions.


Highly regio- and enantioselective synthesis of γ,δ-unsaturated amido esters by catalytic hydrogenation of conjugated enamides.
An efficient and highly regio- and enantioselective catalytic asymmetric hydrogenation of α,γ-dienamido esters to γ,δ-unsaturated amido esters has been achieved using Rh/TangPhos as the catalyst. A series of γ,δ-unsaturated amido acids were furnished in excellent yields with up to 99% ee. This effective methodology was applied in the asymmetric synthesis of key intermediate of Ramipril, an ACE inhibitor.


High performance amorphous ZnMgO/carbon nanotube composite thin-film transistors with a tunable threshold voltage.
Here we report the fabrication and characterization of high mobility amorphous ZnMgO/single-walled carbon nanotube composite thin film transistors (TFTs) with a tunable threshold voltage. By controlling the ratio of MgO, ZnO and carbon nanotubes, high performance composite TFTs can be obtained with a field-effect mobility of up to 135 cm(2) V(-1) s(-1), a low threshold voltage of 1 V and a subthreshold swing as small as 200 mV per decade, making it a promising new solution-processed material for high performance functional circuits. A low voltage inverter is demonstrated with a functional frequency exceeding 5 kHz, which is only limited by parasitic capacitance rather than the intrinsic material speed. The overall device performance of the composite TFTs greatly surpasses not only that of the solution-processed TFTs, but also that of the conventional amorphous or polycrystalline silicon TFTs. It therefore has the potential to open up a new avenue to high-performance, solution-processed flexible electronics which could significantly impact the existing applications, and enable a whole new generation of flexible, wearable, or disposable electronics.


Environmental Impacts and Recovery After the Hebei Spirit Oil Spill in Korea.
The Hebei Spirit oil spill (HSOS) on December 7, 2007 was the worst oil spill recorded in Korea, with the release of approximately 10,900 tons of crude oil and 375 km of coastline polluted along the west coast of Korea. Cleanup operation was conducted by official and contract responders as well as volunteers for massive oil containment and removal of heavy accumulations of stranded oil. Together with the oil cleanup, a long-term environmental impact assessment (EIA) of the HSOS was initiated based on the Marine Environmental Management Act, which covers oil contamination in a multimedia environment, toxic effects on organisms, and ecosystem injury. This review summarizes the long-term monitoring results of HSOS EIA focused on (1) pollution status of seawater, sediment, and bivalves, (2) ecotoxicological effects, and (3) ecosystem recovery. Overall, concentrations of petroleum hydrocarbons in the environment indicated that their concentrations were well down to at or near background or pre-spill contamination levels at most sites after 1 year. The potential toxic effects of residual oils in sediments have decreased to background levels in most coastal areas of Taean. The entire ecosystem in the most affected area of the Taean coasts appear to be considerably, but not fully, recovered at present, namely after 8 years of the HSOS. The presence of lingering oil and elevated contamination levels at several sites still require continuous long-term monitoring.


How and When Perpetrators Reflect on and Respond to Their Workplace Ostracism Behavior: A Moral Cleansing Lens.
This study investigates the association between the previous workplace ostracism of employees and their subsequent helping behavior by drawing on moral cleansing theory in the Chinese context, exploring the mediating roles of employees' guilt and perceived loss of moral credit and the moderating role of their moral identity symbolization. The data were collected from a two-stage time-lagged survey of 284 Chinese employees. Regression analysis and the bootstrapping method are used in this article to examine the theoretical hypotheses. The results indicate that employees' previous ostracism behaviors positively affected their guilt experience and perceived loss of moral credit. Subsequently, the relationship between employees' workplace ostracism and their helping behavior is mediated by guilt experience and perceived loss of moral credit. Furthermore, moral identity symbolization positively moderated the indirect "workplace ostracism-helping behavior" linkage via guilt and perceived loss of moral credits; in other words, for employees who have a higher degree of moral identity symbolization, the mediating effect is more significant, and vice versa. This study does not merely clarify the theoretical relationship between perpetrators' workplace ostracism and their helping behavior, which enriches the explanatory logic of related research on workplace ostracism and the cause of helping behavior, but also expand the application scope of moral cleansing theory. Further, we aim practically to bring enlightenment to human resource management reform, corporate culture construction, and positive behavior management.


Morphological growth dynamics, mechanical stability, and active microtubule mechanics underlying spindle self-organization.
The spindle is a dynamic intracellular structure self-organized from microtubules and microtubule-associated proteins. The spindle's bipolar morphology is essential for the faithful segregation of chromosomes during cell division, and it is robustly maintained by multifaceted mechanisms. However, abnormally shaped spindles, such as multipolar spindles, can stochastically arise in a cell population and cause chromosome segregation errors. The physical basis of how microtubules fail in bipolarization and occasionally favor nonbipolar assembly is poorly understood. Here, using live fluorescence imaging and quantitative shape analysis in Xenopus egg extracts, we find that spindles of varied shape morphologies emerge through nonrandom, bistable self-organization paths, one leading to a bipolar and the other leading to a multipolar phenotype. The bistability defines the spindle's unique morphological growth dynamics linked to each shape phenotype and can be promoted by a locally distorted microtubule flow that arises within premature structures. We also find that bipolar and multipolar spindles are stable at the steady-state in bulk but can infrequently switch between the two phenotypes. Our microneedle-based physical manipulation further demonstrates that a transient force perturbation applied near the assembled pole can trigger the phenotypic switching, revealing the mechanical plasticity of the spindle. Together with molecular perturbation of kinesin-5 and augmin, our data propose the physical and molecular bases underlying the emergence of spindle-shape variation, which influences chromosome segregation fidelity during cell division.


Triple helical collagen-like peptides: engineering and applications in matrix biology.
Collagen, a large insoluble protein with a characteristic triple helical structure, is found as the most prominent component of extracellular matrix. The functions of collagen are not limited to providing mechanical strength to various tissues and organs as a structural protein, as it has been pointed out that collagen exhibits various biological functions through specific interactions with other macromolecules. However, the use of native triple helical collagen is often troublesome because of its insolubility and gelating properties. Instead, triple helical collagen-like peptides have been designed and are used as collagen surrogates in studies on collagen structure, stability, and biological functions including binding to other proteins and cultured cells. This article reviews recent progress in peptide design, synthesis, and the applications of collagen-like peptides in current matrix biology, while emphasizing the advantages of the peptide-based strategy.


Role of nitric oxide and superoxide anions in interleukin-1 beta-induced airway hyperresponsiveness to bradykinin.
We investigated the role of endogenous nitric oxide (NO) and superoxide anions in recombinant human interleukin-1 beta (rhIL-1 beta)-induced bronchial hyperresponsiveness (BHR) and neutrophilia in Brown-Norway rats. Aminoguanidine (100 mg/kg/d) administered subcutaneously for 3 d, an inhibitor of inducible NO synthase, L-arginine (100 mg/kg/d administered subcutaneously for 3 d, a specific precursor for the synthesis of NO, and apocynin (5 mg/kg/orally), an inhibitor of superoxide anion (O2-)-generating NADPH oxidase in macrophages and neutrophils, were administered prior to administration of rhIL-1 beta (500 U) intratracheally. Aminoguanidine in addition to another inhibitor of NO synthase, NW-nitro-L-arginine methyl ester (L-NAME) 100 mg kg/d administered subcutaneously for 3 d augmented bronchial responsiveness to inhaled bradykinin (BK) but not to acetylcholine (ACh), an effect reversed by L-arginine. rhIL-1 beta-treated rats also demonstrated BHR to BK but not to ACh, associated with neutrophilia in bronchoalveolar lavage fluid (BALF). rhIL-1 beta-induced BHR and neutrophilia were neither further increased by aminoguanidine nor inhibited by L-arginine. Apocynin, however, significantly inhibited rhIL-1 beta-induced BHR but not the BALF neutrophilia. Suppression of NO generation and generation of O2- from macrophages and infiltrating neutrophils may be important in rhIL-1 beta-induced airway hyperresponsiveness to bradykinin.


Phylogeography of the neotropical epiphytic orchid, genus Dracula.
The tropical Andes constitute a natural barrier between the Pacific Ocean and the Atlantic; in these mountains, are a great variety of Ecosystems, defined by factors such as orography, winds, humidity, temperature, among others. Some of these Ecosystems have different environmental conditions from tropical ones. In them, there is a great Biodiversity, in some cases endemic and associated with relatively small geographic areas. An example of this biodiversity is the orchids of the genus Dracula, about which discussions are currently generated due to the difficulty in classifying their members. The present work shows a study where DNA was isolated and sequenced from plant samples obtained from 52 species of orchids of the genus Dracula, which were analyzed using the MEGA7 software. Phylogenetic analysis of the DNA sequences showed a well-resolved topology that reflects a geographical pattern of several major clades of the Pacific and Atlantic watersheds. Geophysical conditions of the Andes have generated greater biodiversity of the genus Dracula on the side of the Pacific. Although the species Dracula cordobae and alessandroi reported on both sides of the study site belong to the same clade and show limited mobility through the drier area to the South of the mountain range.


Use of cultured keratinocytes in the treatment of severe burns.
A 6 1/2-year-old child was admitted to the emergency ward for third degree burn injuries representing 40% of body surface. Shock therapy was first applied. After débridement and in a series of operating sessions hands were grafted with full-thickness skin and most other wounds were covered with mesh grafts. On the 14th day after admission a piece of approximately 5 cm2 split thickness scalp skin was used to expand the keratinocytes by cell culture techniques according to the method of Rheinwald and Green (7). After 20 days 4 sheets of cells of first subculture, each 10 cm in diameter, representing a surface of approximately 300 cm2, were implanted on the front of the left thigh, which was burnt third-degree deep. Light microscopy of punch biopsies from mesh grafted and keratinocyte implanted sites taken 5 months after grafting showed a well differentiated epidermis overlying scar tissue. The following conclusions were drawn: Autologous keratinocytes did take on a third degree wound. No basic difference was observed neither clinically nor histologically between mesh-grafted and keratinocyte-implanted sites. The epidermis formed by keratinocyte implantation lacked pigmentation but presented an aesthetically better appearance than the mesh-graft treated sites. Given the scarcity of donor sites and the results obtained by implanting keratinocyte cultures, this latter technique may be resorted to in any extended and deep burn injury.


Leaky gut and the liver: a role for bacterial translocation in nonalcoholic steatohepatitis.
Gut flora and bacterial translocation (BT) play important roles in the pathogenesis of chronic liver disease, including cirrhosis and its complications. Intestinal bacterial overgrowth and increased bacterial translocation of gut flora from the intestinal lumen predispose patients to bacterial infections, major complications and also play a role in the pathogenesis of chronic liver disorders. Levels of bacterial lipopolysaccharide, a component of gram-negative bacteria, are increased in the portal and/or systemic circulation in several types of chronic liver disease. Impaired gut epithelial integrity due to alterations in tight junction proteins may be the pathological mechanism underlying bacterial translocation. Preclinical and clinical studies over the last decade have suggested a role for BT in the pathogenesis of nonalcoholic steatohepatitis (NASH). Bacterial overgrowth, immune dysfunction, alteration of the luminal factors, and altered intestinal permeability are all involved in the pathogenesis of NASH and its complications. A better understanding of the cell-specific recognition and intracellular signaling events involved in sensing gut-derived microbes will help in the development of means to achieve an optimal balance in the gut-liver axis and ameliorate liver diseases. These may suggest new targets for potential therapeutic interventions for the treatment of NASH. Here, we review some of the mechanisms connecting BT and NASH and potential therapeutic developments.


Eyelid abscess arising from an epidermal inclusion cyst.
A 72-year-old woman presented with a 3-day history of focal pain and swelling of the left upper eyelid. A cystic lesion had been present in this area for longer than 7 years, and a lesion consistent with an epidermal inclusion cyst was present on the contralateral upper eyelid. An erythematous, tender, fluctuant mass was noted on the left upper eyelid consistent with abscess. It was drained and cultured, and the infection resolved with oral amoxicillin/clavulanate potassium followed by surgical excision. Histopathologic examination demonstrated an epidermal inclusion cyst and cultures grew anaerobic gram-positive cocci. Epidermal inclusion cysts are common eyelid lesions, yet infection is rare and may result from the spread of normal flora through the pore often found on the anterior surface of these lesions.


Malignant Leydig cell tumor in dogs: two cases and a review of the literature.
Malignant Leydig cell tumor (MLCT) is a rare testicular tumor in dogs. We report herein 2 dogs with MLCT and cutaneous metastasis. Grossly, marked enlargement and distortion of the involved testes were noted; on cut surface, the parenchyma was completely replaced by neoplastic tissue. In addition, these tumors had extensive necrosis and hemorrhage. Case 1 had a rapidly growing cutaneous mass in the left angle of the mouth; the lesion was well-circumscribed and had an indistinct lobular pattern. Case 2 had multiple cutaneous masses in the dorsal neck region, the thoracic back region, and the right hindlimb. Microscopically, the tumor lobules were composed of oval-to-polyhedral cells with eosinophilic cytoplasm and resembled testicular tumors. By immunohistochemistry, the neoplastic cells in both the testicular and cutaneous tumors were positive for inhibin-alpha and melan A. The mitotic counts of the primary tumors from cases 1 and 2 were 21 and 11 per 10 high-power fields, respectively. Based on these findings, the cases were diagnosed as MLCT with cutaneous metastasis. Ki-67 expression in the neoplastic cells of the 2 cases was higher than in benign Leydig cell tumors. Our findings may be helpful for the diagnosis of canine MLCT.


Experimental thyroiditis in guinea pigs and rabbits. Immunization with thyroglobulin and bovine thyroid gland suspension.
Morphological changes in the thyroid glands of the guinea pigs with autoimmune thyroiditis (EAT) experimentally induced by thyroglobulin (TGL) or immunization by the suspension of thyroid gland cells with CFA manifested mainly by atrophy and alterations of follicular cells, fibrotic tissue changes, formation of inflammatory lymphoplasmocytic infiltrations, multiplication of C-cells and by the increase in the proportion of lymphocytes with activated nucleoli in the tissue. The antigenic effects of TGL differed from those of the cell suspension; the effects of TGL participated especially in the formation of the infiltrates, the effects of cell suspension participated in the diapedesis of mononuclears and in the multiplication of C-cells. The findings correspond to the principal findings in human autoimmune lymphocytic thyroiditis. In an electron microscope, strongly dilated cisterns of endoplasmic reticulum (ER) and multiplied mitochondria in the cytoplasm of altered follicular cells were found. The wall of the follicles exhibited fully intact or altered C-cells. The latter had a large number of granules in cytoplasm with an unusually clear medullary substance. In the rabbit thyroid glands no morphological changes were observed following the immunization with both antigens. TGL antibodies examined immunohistochemically in the sera were present in all the sera of guinea pigs immunized with TGL and CFA. The antibodies determined by dot immunodetection were present in the sera of all guinea pigs immunized with TGL+CFA, the titres reached the level of 1:81 to 1:729; the highest titres were observed in the guinea pigs following the immunization by dose of 7.7 mg/kg after 12 weeks of immunization.


Tackling the problem of poor ward cleanliness.
Both the environment and equipment used in health-care facilities have been identified as sources of infection. The cleaning of any hospital environment must be carried out in a pre-planned, clearly structured and defined manner. Domestic staff need thorough basic training and regular updates.


Obesity increases risk of anticoagulation reversal failure with prothrombin complex concentrate in those with intracranial hemorrhage.
Not all patients with warfarin-related acute intracranial hemorrhage (ICH) achieve full reversal of international normalized ratio (INR) after the first dose of weight-based prothrombin complex concentrate (PCC). We sought to identify factors associated with anticoagulation reversal failure after the first dose of PCC. Consecutive patients who were hospitalized with warfarin-related acute ICH at a tertiary center between 1 January 2010 and 31 December 2012 were studied. Anticoagulation reversal failure was defined as INR ≥ 1.5 after the first dose of PCC. Logistic regression was performed to determine the predictors of anticoagulation reversal failure. Fifty-one patients with acute ICH received PCC for warfarin reversal using a weight-based protocol. Overall, 23 (45%) patients did not achieve full reversal of INR after the first dose. Those with anticoagulation reversal failure were obese (body mass index > 30 kg/m(2)) (41% vs. 14%, p = 0.03), had a higher initial INR (3.0 ± 1.4 vs. 2.0 ± 0.7, p = 0.001), and had a higher prevalence of initial INR >2.0 (22% vs. 67%, p = 0.001), compared with those who were successfully reversed. Multivariable logistic regression identified obesity (odds ratio 7.88, 95% CI 1.12 to 55.68) and initial INR >2.0 (odds ratio 12.49, 95% CI 2.27 to 68.87) as independent predictors of anticoagulation reversal failure. Obesity and elevated initial INR are independently associated with anticoagulation reversal failure using the weight-based PCC protocol in patients with warfarin-related acute ICH. Further studies are needed to determine more effective dosing protocols and individualized strategies for anticoagulation reversal after acute ICH, especially among obese patients.


Prolactin response to D-fenfluramine challenge test as a predictor of treatment response to haloperidol in acute schizophrenia.
D-fenfluramine has been identified as a highly selective serotonin (5-HT) releaser and re-uptake inhibitor. The objective of our study was to investigate prolactin response to D-fenfluramine challenge in non-medicated, first episode schizophrenics. We hypothesized that 5-HT reactivity can predict a response to the neuroleptic treatment. Twenty-three inpatients, 11 males and 12 females, at the Prague Psychiatric Center participated in the study. Inclusion criteria were: ICD-10 diagnosis of schizophrenia, first episode or duration of illness shorter than 36 months. D-fenfluramine challenge test was performed before 4 weeks of the haloperidol treatment. During the test, prolactin plasma levels were measured. The Brief Psychiatric Rating Scale (BPRS) was administered before and after the treatment. A statistically significant negative correlation was found between prolactin response to the D-fenfluramine challenge and improvement of psychopathology measured by the change in total BPRS score (p = 0.0004), in positive (p = 0.0403), negative (p = 0.0267), and anxiety-depression symptoms of BPRS (p = 0.0014). Our data support the original hypothesis that there is a relationship between 5-HT system activity and treatment response. The higher responsiveness of the 5-HT system in first episode, non-medicated schizophrenics, was associated with a poorer treatment response to haloperidol, an antidopaminergic neuroleptic.


Thromboembolism in patients with atrial fibrillation.
On retrospective review of records of 150 patients with atrial fibrillation (AF), 31% experienced a stroke or peripheral embolism. The AF was not associated with cardiac valvular disease in 80% of the 150 patients. Most of the cerebral infarcts were large, disabling, and unheralded by transient ischemic attack. The thromboembolism typically occurred in patients whose AF was undetected prior to the infarction. Half of the patients with an ischemic event suffered multiple events, with one fourth of the recurrences arising within two weeks.


Enhanced [3H]DOPA and [3H]dopamine turnover in striatum and frontal cortex in vivo linked to glutamate receptor antagonism.
We tested the hypothesis that blockade of NMDA glutamate receptors in brain enhances dopamine turnover. We blocked this class of glutamate receptors in the rat brain in vivo with dizocilpine (MK-801) and measured the accumulation of radiolabeled DOPA and its metabolites as functions of time after intravenous bolus injection. Using the time courses of the accumulated metabolites, we calculated the turnover constants of enzymes mediating dopamine synthesis and catabolism. Dizocilpine treatment for 8 days enhanced the rates of DOPA decarboxylation and dopamine oxidation (monoamine oxidation) 4- and 16-fold, respectively, in neostriatum and 10- and 3-fold, respectively, in frontal cortex. The findings are not inconsistent with the hypothesis that the psychotomimetic properties of dizocilpine may be the manifestation of denervation hypersensitivity linked to activation of key enzymes of dopamine turnover in striatum.


Development of plant-produced protein body vaccine candidates for bluetongue virus.
Bluetongue is a disease of domestic and wild ruminants caused by bluetongue virus serotypes (BTV), which have caused serious outbreaks worldwide. Commercially available vaccines are live-attenuated or inactivated virus strains: these are effective, but there is the risk of reversion to virulence or reassortment with circulating strains for live virus, and residual live virus for the inactivated vaccines. The live-attenuated virus vaccines are not able to distinguish naturally infected animals from vaccinated animals (DIVA compliant). Recombinant vaccines are preferable to minimize the risks associated with these vaccines, and would also enable the development of candidate vaccines that are DIVA-compliant. In this study, two novel protein body (PB) plant-produced vaccines were developed, Zera®-VP2ep and Zera®-VP2. Zera®-VP2ep contained B-cell epitope sequences of multiple BTV serotypes and Zera®-VP2 contained the full-length BTV-8 VP2 codon-optimised sequence. In addition to fulfilling the DIVA requirement, Zera®-VP2ep was aimed at being multivalent with the ability to stimulate an immune response to several BTV serotypes. Both these candidate vaccines were successfully made in N. benthamiana via transient Agrobacterium-mediated expression, and in situ TEM analysis showed that the expressed proteins accumulated within the cytoplasm of plant cells in dense membrane-defined PBs. The peptide sequences included in Zera®-VP2ep contained epitopes that bound antibodies produced against native VP2. Preliminary murine immunogenicity studies showed that the PB vaccine candidates elicited anti-VP2 immune responses in mice without the use of adjuvant. These proof of concept results demonstrate that Zera®-VP2ep and Zera®-VP2 have potential as BTV vaccines and their development should be further investigated.


Immunophenotype of vascular rejection in renal transplants.
Understanding of the pathogenesis of vascular rejection processes encountered in renal transplants is limited. Although initially and still widely thought to be antibody mediated, it is commonly difficult to demonstrate deposition of immunoglobulin (Ig) in affected arteries. We studied the vascular lesions present in 22 transplanted human kidneys with a panel of antibodies and lectins to evaluate the presence of granulocytes (Leu M1), leukocytes (anti-CD45), B cells (L26), T cells (UCHL-1), monocyte/macrophages (HAM 56), endothelial proliferation (Ulex I factor VIII-related antigen), and smooth muscle proliferation (HHF 35). Active (cellular) vascular rejection showed intimal infiltration of T lymphocytes and monocytes/macrophages (Mac) but not B lymphocytes. Lesions of greater chronicity (reduction in cellularity, increase in intimal stromal matrix) showed progressive diminution of the T cell infiltrate but persistence of Mac accompanied by increased smooth muscle cells. Endothelial alterations were limited to disruption and lifting from supporting stroma by infiltrating inflammatory cells; proliferative changes as detected by increased numbers of cells binding Ulex I were not identified. The cell infiltrates were similar in large (renal artery) and small (interlobular) arteries. Evidence for specific deposition of Ig was not present in cases studied by immunofluorescence studies. These studies suggest vascular rejection is commonly mediated by cellular immune mechanisms and the general supposition equating vascular rejection occurring beyond the peritransplant period with humoral rejection is mistaken. Persistent Mac in chronic lesions may be limited to scavenger functions, but their presence suggests activity in modulating intimal proliferation analogous to current hypotheses for such a role for Mac in atherosclerosis.


Comparison of the effectiveness of whole-brain radiotherapy plus temozolomide versus whole-brain radiotherapy in treating brain metastases based on a systematic review of randomized controlled trials.
Temozolomide (TMZ) combination with whole-brain radiotherapy (WBRT) has been tested by many randomized controlled trials in the treatment of brain metastases (BMs) in China and other countries. We performed an up-to-date meta-analysis to determine (i) the log odds ratios (LORs) of objective response (ORR) and adverse effects (AEs) for all-grade, and (ii) the T value of mean overall survival in patients with BMs treated with WBRT combined with TMZ versus WBRT alone. PubMed, Chinese National Knowledge Infrastructure, and WanFang Data were searched for articles published up to 28 January 2015. Eligible studies were selected according to the PRISMA statement. ORR, AEs, and 95% confidence intervals were calculated using random-effects models. Eighteen studies were included in our analysis. A total of 1028 participants were enrolled. Summary LORs of ORR were 1.0239 (P<0.0001) on comparing WBRT plus TMZ with WBRT ORR (n=17). The overall mean difference of mean overall survival (n=17) between TMZ plus WBRT and WBRT was 2.2505 weeks (P=0.02185). There was a significant difference between WBRT plus TMZ and WBRT alone with a LOR of AEs for all-grade of (i) 0.923 for gastrointestinal toxicity and (ii) 0.7978 for myelosuppression. Sensitivity analysis and subgroup analysis were also performed. The 18 eligible randomized controlled trials demonstrated that the combination of WBRT and TMZ significantly improves the ORR and is statistically insignificant in prolonging the survival of patients with BMs. In addition, an increase in the incidence of gastrointestinal toxicity and myelosuppression was significant for all-grade.


Progresses and challenges of utilizing traditional birth attendants in maternal and child health in Nigeria.
Despite advances in modern healthcare, Traditional Birth Attendants (TBA) have continued to be heavily utilized in rural communities in Nigeria. Major disparities in maternal health care in Nigeria remain present despite the goal of the United Nations Millennium Development Goal to reduce maternal mortality by 2015. The objective of this study is to review the contribution of TBAs in the birthing process in Nigeria, and to examine barriers and opportunities for utilizing TBAs in improving maternal and child health outcomes in Nigeria. A literature review of two major electronic databases was conducted using the PRISMA framework to identify English language studies conducted between 2006 and 2016. Inclusion criteria included articles that examined the role of traditional birth attendants as a factor influencing maternal health in Nigeria. The value of TBAs has not been fully examined as few studies have aimed to examine its potential role in reducing maternal mortality with proper training. Eight manuscripts that were examined highlighted the role of TBAs in maternal health including outcomes of utilizing trained versus non-trained TBAs. Specific areas of training for TBAs that were identified and recommended in review including: recognizing delivery complications, community support for TBA practices through policy, evaluation of TBA training programs and increasing collaboration between healthcare facilities and TBAs. Policies focused on improving access to health services and importantly, formal health education training to TBAs, are required to improve maternal health outcomes and underserved communities.


Segregational respiratory-deficient mutants of a "petite negative" yeast Schizosaccharomyces pombe 972h-.
No viable respiratory-deficient mutants of Schizosaccharomyces pombe 972h(-) could be obtained by acriflavine and ethidium bromide treatments. These mutagens induce 15 to 70% of microcolonies which, after a growth-lag of a few days, further develop into normal, respiratory-competent colonies. These results suggest that unstable petites were induced. Segregational respiratory-deficient mutants resistant to cobalt sulfate inhibition were isolated. Some of these strains are deficient in cytochrome a + a(3) and respire at low rates. The morphology of their mitochondrial membranes is modified: either the cristae are absent or they show aberrant concentric or tubular structures. Segregational mutants resistant to the respiratory inhibitors, 2,4-dinitrophenol or decamethylene diguanidine, were obtained. Neither mitochondrial structure nor function seems to be modified in these mutants. A segregational mutant resistant to benzimidazole inhibition does not grow on glycerol, although neither growth on glucose nor respiration appear to be affected.


[Influence of MRI on the T, N staging system of nasopharyngeal carcinoma].
To investigate the influence of MRI on the T, N staging system, 5(th) edition, UICC and the 92 staging system, China by comparing CT and MRI of the nasopharyngeal carcinoma (NPC). From 1993 to Dec. 2000, fifty-six NPC patients proved by pathology underwent CT and MRI examination with Philip T5-II ultra-magnetic system (0.5T) and Elscient CT twin flash. Routine axial scans by CT and SE sequence with axial, sagittal and coronal scans by MRI from oral pharynx (lower border of second cervical vertebra) to supracellar cistern were done with enhancement (50/56). The tumor beyond the nasopharyngeal cavity was accurately defined because the pharyngobasilar fascia could be seen by MRI which appeared to be more sensitive than CT in revealing the invaded soft tissues surrounding the nasopharyngeal cavity, such as longus colli (14 by CT and 26 by MRI), tensor veli palatini and levator veli palatini (17 by CT and 42 by MRI), the skull base erosion (15 by CT and 23 by MRI) and enlargement of retropharyngeal lymph nodes (13 by CT and 24 by MRI). As a result, 28.6% (16/56) of NPC staging system, UICC and 33.9% (19/56) of the 92 staging system of China should undergo changes. MRI is able to reveal the invasion extent into the structures around the tumor mass more accurately than CT, with the pharyngobasilar fascia readily seen and the infiltration and/or destruction of the skull base more easily visualized by MRI than by CT. The impact of MRI upon the 92 staging system of China lies in the differentiation of direct infiltration by the tumor from the enlargement of the retropharyngeal lymph node and the early detection of skull base erosion. The Influence of MRI on the staging system of NPC is more pronounced than that of CT.


Evaluation and analysis of formation of bone at the palate in patients with cleft lip and palate after palatoplasty based on computed tomograms and three-dimensional data.
There are various techniques for palatoplasty, but no studies of postoperative osteogenesis at the palatal fissure. In the cranial and maxillofacial region it is thought to develop from the periosteum, so palatoplasty with mucoperiosteal flaps may encourage new bone to form at the fissure. We evaluated the status of osteogenesis in the hard palate after palatoplasty on computed tomograms (CT). We studied 29 patients (22 boys and 7 girls) with unilateral cleft lip and palate who had pushback palatoplasty with the use of CT obtained between May 2003 and March 2007. Age at the time of operation was recorded. The width of the palatal fissure at the first premolar, the first molar, and the maxillary posterior region were measured on coronal CT. The mean (SD) age at the time of palatoplasty was 16 (2) months. The mean (SD) width of the fissure at the first molar was 3.96 (3.1) mm, and bony union was seen in four patients. The width of the fissure was significantly less at the first molar than at the other sites (p = 0.006). The shape of the margin of the fissure was irregular in nearly all patients. The width of the fissure at the first molar became significantly less, suggesting that osteogenesis had occurred. In some patients the height of the fissure differed. Given the results of previous studies, bony regeneration from the periosteum most likely happens together with regeneration from the margins of the fissure.


Protection and antibody reactivity following vaccination of lumpfish (Cyclopterus lumpus L.) against atypical Aeromonas salmonicida.
Atypical Aeromonas salmonicida is frequently associated with disease and mortality in farmed lumpfish (Cyclopterus lumpus L). Challenge experiments using different modes of exposure identified both high and low pathogenic isolates. Intraperitoneal vaccination induced production of high levels of specific antibodies particularly in fish given multiple injections. The immune sera contained antibodies cross reactive with both high and low pathogenic isolates. SDS-PAGE and LC/MSMS analyses showed that the highly virulent isolate expressed the virulence array protein (A-layer) while the less virulent isolate did not. Vaccines, containing the highly virulent isolate, formulated as a monovalent or as a trivalent vaccine, provided 73 and 60 relative percent survival (RPS) respectively, following intraperitoneal challenge. The detection of high levels of specific antibodies in immune sera and the protection provided by the test vaccines strongly indicate that it is possible to vaccinate lumpfish against atypical A. salmonicida and most probably also against other infectious bacterial diseases.


A formylpeptide receptor, FPRL1, acts as an efficient coreceptor for primary isolates of human immunodeficiency virus.
More than 10 members of seven-transmembrane G protein-coupled receptors (GPCRs) have been shown to work as coreceptors for human immunodeficiency virus type 1 (HIV-1), HIV type 2 (HIV-2), and simian immunodeficiency viruses (SIVs). As a common feature of HIV/SIV coreceptors, tyrosine residues are present with asparagines, aspartic acids or glutamic acids in the amino-terminal extracellular regions (NTRs). We noticed that a receptor for N-formylpeptides, FPRL1, also contains two tyrosine residues accompanied by glutamic acids in its NTR. It was reported that monocytes expressing CCR5 and FPRL1 in addition to CD4 are activated by treatment with ligands or agonists of FPRL1. Activated monocytes down-modulate CCR5 and become resistant to infection by HIV-1 strains. Thus, FPRL1 plays important roles in protection of monocyptes against HIV-1 infection. However, its own coreceptor activity has not been elucidated yet. In this study, we examined coreceptor activities of FPRL1 for HIV/SIV strains including primary HIV-1 isolates. A CD4-transduced human glioma cell line, NP-2/CD4, is strictly resistant to HIV/SIV infection. We have reported that when NP-2/CD4 cells are transduced with a GPCR having coreceptor activity, the cells become susceptible to HIV/SIV strains. When NP-2/CD4 cells were transduced with FPRL1, the resultant NP-2/CD4/FPRL1 cells became markedly susceptible to some laboratory-adapted HIV/SIV strains. We found that FPRL1 is also efficiently used as a coreceptor by primary HIV-1 isolates as well as CCR5 or CXCR4. Amino acid sequences linked to the FPRL1 use could not be detected in the V3 loop of the HIV-1 Env protein. Coreceptor activities of FPRL1 were partially blocked by the forymyl-Met-Leu-Phe (fMLF) peptide. We conclude that FPRL1 is a novel and efficient coreceptor for HIV/SIV strains. FPRL1 works as a bifunctional factor in HIV-1 infection. Namely, the role of FPRL1 in HIV-1 infection is protective and/or promotive in different conditions. FPRL1 has been reported to be abundantly expressed in the lung, spleen, testis, and neutrophils. We detected mRNA expression of FPRL1 in 293T (embryonal kidney cell line), C8166 (T cell line), HOS (osteosarcoma cell line), Molt4#8 (T cell line), U251MG (astrocytoma cell line), U87/CD4 (CD4-transduced glioma cell line), and peripheral blood lymphocytes. Roles of FPRL1 in HIV-1 infection in vivo should be further investigated.


Brownian dynamics simulations of two-dimensional model for hopping times.
Brownian dynamics simulations are used to study the mean first passage time for a hard disk diffusing in a channel. The disk has to hop pass another disk with the same diameter sigma but fixed in contact with the wall. This mean first passage time is expected to diverge with an exponent eta as the channel width (2R(p)) approaches that of the nonpassing limit (2sigma) for one disk to hop pass another. Our one disk model is similar to a disk diffusing in a two-dimensional container, shaped as a simplified configuration space of two passing disks in a flat channel of Kalinay. The Kalinay model was proposed recently to elucidate the disagreements between two different theoretical predictions for the exponent eta of the two disk hopping time divergences. For the one disk model, simple transition state theory and a Fick-Jacobs type of dimensional reduction approach predict exponents of 1 and 12, respectively. Our Brownian dynamics simulation, results are consistent with a value of 1.


Endobronchial Ultrasound-guided Intratumoral Injection of Cisplatin for the Treatment of Isolated Mediastinal Recurrence of Lung Cancer.
Isolated hilar and mediastinal recurrence (IMHR) following external beam radiation therapy (EBRT) in patients with lung cancer is common. These patients do not have many treatment options and are usually offered palliative chemotherapy or best supportive care. Endobronchial ultrasound (EBUS)-guided intratumoral injection of cisplatin (ITC) is a novel approach for these patients. The procedure is performed under conscious sedation. The lesion is located with a bronchoscopy using EBUS, and a 22-gauge EBUS needle is advanced through the working channel of the scope and locked in position. Under ultrasound guidance, the wall of the tracheobronchial tree is punctured and the needle is moved into the target lesion. The needle stylet is then removed and cisplatin (40 mg/40 mL) is injected into the lesion. One to two sites are treated per session. Details of the procedure are described in the protocol section of paper. At our center, 50 sites were treated in 36 patients (19 males, 17 females). The mean age of our cohort was 61.9 ±8.5 years. We performed final analyses on 35 patients and 41 sites. 24/35 (69%) had complete or partial response (responders), whereas 11/35 (31%) had stable or progressive disease (non-responders). Overall, survival in our group was 8 months (95% CI of 6-11 months), with patients who responded having significantly better survival than the ones who did not.


Effects of total glucosides of paeony for delaying onset of Sjogren's syndrome: an animal study.
To investigate the effectiveness of total glucosides of paeony (TGP) on Sjogren's syndrome (SS) using non-obese diabetic (NOD) mice model. Twenty-seven 8-week-old female NOD mice were assigned into TGP group, hydroxychloroquine (HCQ) group and normal saline (NS) group, receiving corresponding drugs respectively and sacrificed at 24-week-old. Saliva flow rate (SFR), ration of regulatory T cells, level of anti-SSA/SSB, histological changes in submandibular glands (SMG) and microarray analysis were assessed. The data were analyzed using SPSS. Compared to NS group, in TGP group, SFR, SMG index and the ration of regulatory T cells were significantly higher, while anti-SSA/SSB and lymphocytic foci were significantly lower. HCQ group demonstrated similar results except SMG index. Altered gene expression was found in 10.71% of TGP and 13.09% of HCQ of the profile. TGP demonstrated a similar effectiveness as HCQ in delaying the onset of SS-like disease in NOD mice.


Oxacillinases and antimicrobial susceptibility of Ralstonia pickettii from pharmaceutical water systems in Croatia.
This study evaluated antibiotic susceptibility and presence of blaOXA22 and blaOXA60 genes in 81 isolates of Ralstonia pickettii obtained from different purified and ultra-pure water systems in two different geographical areas of Croatia. E-test and disc diffusion test were performed to determine antibiotic susceptibility. Polymerase chain reaction was applied to detect genes encoding OXA-22 and OXA-60 oxacillinases previously identified in R. pickettii. The isolates were genotyped by pulsed-field gel electrophoresis. The results revealed variable susceptibility/resistance profiles. Our isolates exhibited high susceptibility rates to ceftriaxone, cefotaxime, piperacillin-tazobactam, ciprofloxacin, imipenem, cefepime and in lesser extent to ceftazidime. High rates of susceptibility were also observed for sulphamethoxazole-trimethoprim and piperacillin. High resistance rates were noticed for ticarcillin-clavulanate, aztreonam and meropenem, as well as for all aminoglycosides tested. Modified Hodge test was positive in 51·9% strains, indicating production of carbapenemases. blaOXA22 and blaOXA60 genes were detected in 37·0 and 80·3% strains, respectively. Pulsed-field gel electrophoresis identified three major clusters containing subclusters. R. pickettii should be taken seriously as a possible cause of nosocomial infections to ensure adequate therapy, to prevent the development of resistant strains and to try to reduce the possibility of R. pickettii surviving in clean and ultra clean water systems.


Isolation and characterization of mu-calpain, m-calpain, and calpastatin from postmortem muscle. I. Initial steps.
Evidence has indicated that mu-calpain, m-calpain, and calpastatin have important roles in the proteolytic degradation that results in postmortem tenderization. Simple assays of these 3 proteins at different times postmortem, however, has shown that calpastatin and mu-calpain both rapidly lose their activity during postmortem storage, so that proteolytic activity of mu-calpain is nearly zero after 3 d postmortem, even when assayed at pH 7.5 and 25 degrees C, and ability of calpastatin to inhibit the calpains is 30% or less of its ability when assayed at death. m-Calpain, however, retains much of its proteolytic activity during postmortem storage, but the Ca(2+) requirement of m-calpain is much higher than that reported to exist in postmortem muscle. Consequently, it is unclear how the calpain system functions in postmortem muscle. To clarify this issue, we have initiated attempts to purify the 2 calpains and calpastatin from bovine semitendinosus muscle after 11-13 d postmortem. The known properties of the calpains and calpastatin in postmortem muscle have important effects on approaches that can be used to purify them. A hexyl-TSK hydrophobic interaction column is a critical first step in separating calpastatin from the 2 calpains in postmortem muscle. Dot-blot assays were used to detect proteolytically inactive mu-calpain. After 2 column chromatographic steps, 5 fractions can be identified: 1) calpastatin I that does not bind to an anion-exchange matrix, that does not completely inhibit the calpains, and that consists of small polypeptides <60 kDa; 2) calpastatin II that binds weakly to an anion-exchange matrix and that contains polypeptides <60 kDa; all these polypeptides are smaller than the native 115- to 125-kDa skeletal muscle calpastatin; 3) proteolytically active mu-calpain even though very little mu-calpain activity can be detected in zymogram assays of muscle extracts from 11- to 13-d postmortem muscle; this mu-calpain has an autolyzed 76-kDa large subunit but the small subunit consists of 24-, 26- and a small amount of unautolyzed 28-kDa polypeptides; 4) proteolytically active m-calpain that is not autolyzed; and 5) proteolytically inactive mu-calpain whose large subunit is autolyzed to a 76-kDa polypeptide and whose small subunit contains polypeptides similar to the proteolytically active mu-calpain. Hence, loss of calpastatin activity in postmortem muscle is due to its degradation, but the cause of the loss of mu-calpain activity remains unknown.


Development of a Chinese childbearing attitude questionnaire for infertile women receiving in vitro fertilization treatment.
The purpose of this study was to report the second phase of instrument development, a culturally sensitive questionnaire of childbearing attitudes to assess the psychosocial responses of infertile women. Using a nonexperimental quantitative design, we investigated 238 women who are undergoing in vitro fertilization treatment. Data collection and relevant planning occurred in two phases: in-depth interviews of women to generate items for the questionnaire and establishing the questionnaire's content and construct validity. Through factor analysis, five factors were extracted from the "attitude toward childbearing questionnaire": gender identification with self and society, insurance of marriage and inheritance, happy family life, spiritual investment, and continuing the family line and procreation. The total variance of these five factors was 64.31%. Cronbach's α and test-retest reliability were between .72 and .87 and between .60 and .76, respectively, demonstrating acceptable internal consistency and stability. The information obtained through the questionnaire could be used to provide infertile women with personal counseling and appropriate psychological support during and after assisted reproductive technology.


Limited hydrolysis of the polypeptide chain elongation factor Tu by trypsin. Isolation and characterization of the polypeptide fragments.
The digestion of EF-Tu-GDP (or EF-Tu-GTP) by trypsin [EC 18.104.22.168] under native conditions has been shown to proceed through two different and characteristic stages. 1. In the first phase, the protein is transformed into a fragment (Fragment A) with a molecular weight of 39,000 by exposure to trypsin for a relatively short period of time. Fragment A is unable to catalyze the binding of aminoacyl-tRNA to ribosomes. The ability to promote two partial steps of the binding reaction, i.e., formation of the aminoacyl-tRNA-EF-Tu-GTP ternary complex as well as the methanol-stimulated, ribosome dependent GTPase reaction, was rapidly destroyed. On the other hand, the ability to interact with guanine nucleotides as well as EF-Ts survived well during prolonged digestion. 2. In the second phase of digestion, a nick is introduced in Fragment A to yield two subfragments (Fragments B and C). These two fragments exist as a hybrid molecule which migrates as a single peak on a Sephadex G-75 column, and which dissociates into Fragments B and C only in the presence of 6 M guanidine hydrochloride or 5% sodium dodecyl sulfate. The molecular weights of Fragments B and C, as determined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate, were 22,000 and 12,000 respectively. The hybrid molecule still retained one mole of bound guanine nucleotide and was resistant to further tryptic digestion. 3. Three sulfhydryl groups of EF-Tu were found to be present in Fragment B, both by amino acid analysis of the purified fragments and also by electrophoresis of tryptic digests labeled with N-ethyl[14C]maleimide. 4. The tryptic digestion of EF-Tu-GDP (or EF-Tu-GTP) labeled with N-(1-anilinonaphthyl-4)maleimide (ANM) at SH2 (the second SH), caused a 30% decrease in the fluorescence emission during the first rapid phase of digestion. This indicates that destruction of the hydrophobic environment near SH2 of EF-Tu occurred in the early phase of tryptic digestion. 5. The kinetic studies on the reaction of ANM with EF-Tu before and after tryptic digestion indicated that both Fragment A and the hybrid molecule reacted with ANM in the presence of GTP three to four times more rapidly than in the presence of GDP. Thus, it appears that the ability to induce conformational transition near SH2 by a change of nucleotide ligands is still retained in the hybrid molecule consisting of Fragments B and C.


Identification and Differentiation of Wide Edible Mushrooms Based on Lipidomics Profiling Combined with Principal Component Analysis.
Mushroom, as a kind of higher fungus, is a precious homology resource of medicine and foods. In this study, total lipids were extracted from eight wild edible mushrooms and subsequently characterized by ultra-high-performance liquid chromatography-Quadrupole-Exactive Orbitrap mass spectrometry. 20 lipid classes and 173 molecular species were identified and quantified. Lipid molecules and their concentrations in Boletus speciosus, Boletus bainiugan, and Tricholoma matsutake exhibited significantly different behaviors compared with the remaining mushrooms. Hierarchical cluster analysis revealed that lipid profiles of B. bainiugan were most similar to B. speciosus followed by T. matsutake, Canthar-ellus cibarius, Sarcodon aspratu, Termitomyces eurrhizus, Laccaria laccata, and Thelephora ganbajun. In addition, several differential lipids can be considered as potential biomarkers to distinguish different mushroom species, for instance, lysophosphatidylethanolamine (16:1) and ceramide non-hydroxy fatty acid-dihydrosphingosine (d23:0-10:0). This study provided a new perspective to discriminate the mushroom species from the perspective of lipidomics.


Identification and first insights into the structure of chitin from the endemic freshwater demosponge Ochridaspongia rotunda (Arndt, 1937).
Studies on the identification, properties and function of chitin in sponges (Porifera), which are recognized as the first multicellular organisms on Earth, continue to be of fundamental scientific interest. The occurrence of chitin has so far been reported in 21 marine sponge species and only in two inhabiting fresh water. In this study, we present the discovery of α-chitin in the endemic demosponge Ochridaspongia rotunda, found in Lake Ohrid, which dates from the Tertiary. The presence of chitin in this species was confirmed using special staining, a chitinase test, FTIR, Raman and NEXAFS spectroscopy, and electrospray ionization mass spectrometry (ESI-MS). In contrast to the case of marine sponges, chitin in O. rotunda has been found only within its holdfast, suggesting a role of chitin in the attachment of the sponge to the hard substratum. Isolated fibrous matter strongly resemble the shape and size of the sponge holdfast with membrane-like structure.


In vivo EPR pharmacokinetic evaluation of the redox status and the blood brain barrier permeability in the SOD1G93A ALS rat model.
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder affecting the motor pathways of the central nervous system. Although a number of pathophysiological mechanisms have been described in the disease, post mortem and animal model studies indicate blood-brain barrier (BBB) disruption and elevated production of reactive oxygen species as major contributors to disease pathology. In this study, the BBB permeability and the brain tissue redox status of the SOD1G93A ALS rat model in the presymptomatic (preALS) and symptomatic (ALS) stages of the disease were investigated by in vivo EPR spectroscopy using three aminoxyl radicals with different cell membrane and BBB permeabilities, Tempol, 3-carbamoyl proxyl (3CP), and 3-carboxy proxyl (3CxP). Additionally, the redox status of the two brain regions previously implicated in disease pathology, brainstem and hippocampus, was investigated by spectrophotometric biochemical assays. The EPR results indicated that among the three spin probes, 3CP is the most suitable for reporting the intracellular redox status changes, as Tempol was reduced in vivo within minutes (t1/2 =2.0±0.5min), thus preventing reliable kinetic modeling, whereas 3CxP reduction kinetics gave divergent conclusions, most probably due to its membrane impermeability. It was observed that the reduction kinetics of 3CP in vivo, in the head of preALS and ALS SOD1G93A rats was altered compared to the controls. Pharmacokinetic modeling of 3CP reduction in vivo, revealed elevated tissue distribution and tissue reduction rate constants indicating an altered brain tissue redox status, and possibly BBB disruption in these animals. The preALS and ALS brain tissue homogenates also showed increased nitrilation, superoxide production, lipid peroxidation and manganese superoxide dismutase activity, and a decreased copper-zinc superoxide dismutase activity. The present study highlights in vivo EPR spectroscopy as a reliable tool for the investigation of changes in BBB permeability and for the unprecedented in vivo monitoring of the brain tissue redox status, as early markers of ALS.


Purification and properties of a thiol protease from rat liver nuclei.
A thiol protease was purified about 800-fold from the chromatin fraction of rat liver by employing Sepharose 6B gel filtration, chromatofocusing and Sephadex G-100 gel filtration. It was nearly homogeneous on sodium dodecyl sulfate/polyacrylamide gel electrophoresis and its molecular weight was about 29000. The isoelectric point of the enzyme was 7.1. The pH optimum for degradation of 3H-labelled ribosomal proteins was 4.5. It is noticeable that the maximal activity was shifted to pH 5.5 by DNA, and that 30-40% of the maximal activity was observed at neutral pH in the presence of DNA. The activity was increased about twice by 2-4 mM dithiothreitol. The protease may be specific for the nuclei because it is different from all lysosomal thiol proteases ever known.


A MIXTURE OF NALBUPHINE, AZAPERONE, AND MEDETOMIDINE FOR IMMOBILIZING RINGTAILS (BASSARISCUS ASTUTUS).
We evaluated a combination of nalbuphine HCl (40 mg/mL), azaperone tartrate (10 mg/mL), and medetomidine HCl (10 mg/mL), a combination known as NAM or NalMed-A, in 23 ringtails (Bassariscus astutus) during 29 handling events for a radio-collaring study in southern Oregon, US, from August 2020 to March 2022. The combination was delivered to ringtails by hand injection at 0.075 mL NAM per estimated 1 kg body mass. The mean (± standard deviation, SD) dosage calculated post hoc was 3.366 (±0.724) mg/kg nalbuphine, 0.841 (±0.181) mg/kg medetomidine, and 0.841 (±0.181) mg/kg azaperone. All captured ringtails were effectively immobilized with a mean (SD) induction time of 13.24 (±3.57) min. The medetomidine and nalbuphine components were antagonized with a combination of atipamezole and naltrexone HCl with a mean (SD) recovery time of 2.48 (±1.94) min. This combination appeared to be safe and effective for immobilizing ringtails with a low volume dose, smooth antagonism, and rapid recovery. In addition, NAM does not contain any drugs that are US Drug Enforcement scheduled, which makes it useful for immobilization procedures by wildlife professionals in the US.


Diallyl disulfide increases CDKN1A promoter-associated histone acetylation in human colon tumor cell lines.
Diallyl disulfide (DADS) is an organosulfur compound from garlic, which inhibits colon tumor cell proliferation. In a previous study, we have shown that in Caco-2 and HT-29 cells DADS (200 microM) increases global histone acetylation, CDKN1A mRNA, and p21(waf1) protein levels and induces G2/M cell cycle arrest. These results suggested that DADS could inhibit cell proliferation through at least in part a transcriptional activation of CDKN1A expression involving histone acetylation. In this study, using chromatin immunoprecipitation assays, we demonstrate that in Caco-2 and HT-29 cells histone H4 and/or H3 acetylation is increased within CDKN1A promoter after 3 and 6 h treatments with DADS. These results strongly suggest that histone acetylation, a molecular mechanism implicated in the regulation of gene expression, could account for the induction of CDKN1A expression and the antiproliferating effects of DADS in colon tumor cells.


Efficacy and safety of de-escalation therapy to ertapenem for treatment of infections caused by extended-spectrum-β-lactamase-producing Enterobacteriaceae: an open-label randomized controlled trial.
Carbapenem antibiotics are considered the treatment of choice for serious extended-spectrum beta-lactamase (ESBL)-producing Gram-negative bacteria (GNB) infections. The study objectives were to evaluate efficacy and safety of de-escalation therapy to ertapenem for treatment of infections caused by extended-spectrum-β-lactamase-producing Enterobacteriaceae. We conducted a randomized controlled trial of adult patients with documented ESBL-producing Enterobacteriaceae infections who had received any group 2 carbapenem for less than 96 h. In the intervention group, the previously-prescribed group 2 carbapenem was de-escalated to ertapenem. In the control group, the group 2 carbapenem was continued. During June 2011-December 2014, 32 patients were randomized to the de-escalation group and 34 to the control group. Most common sites of infection were urinary tract infection (42%). Characteristics of both groups were comparable. By using a 15% predefined margin, ertapenem was non-inferior to control group regarding the clinical cure rate (%Δ = 14.0 [95% confidence interval: -2.4 to 31.1]), the microbiological eradication rate (%Δ = 4.1 [-5.0 to 13.4]), and the superimposed infection rate (%Δ = -16.5 [-38.4 to 5.3]). Patients in the de-escalation group had a significantly lower 28-day mortality rate (9.4% vs. 29.4%; P = .05), a significantly shorter median length of stay (16.5 days [4.0-73.25] vs. 20.0 days [1.0-112.25]; P = .04), and a significantly lower defined daily dose of carbapenem use (12.9 ± 8.9 vs. 18.4 ± 12.6; P = .05). Ertapenem could be safely used as de-escalation therapy for ESBL-producing Enterobacteriaceae infections, once the susceptibility profiles are known. Future studies are needed to investigate ertapenem efficacy against ESBL-producing Enterobacteriaceae pneumonia to determine its applicability in life-threatening conditions. ClinicalTrials.gov identifier: NCT01297842 . Registered on 14 February 2011. First patient enrolled on 27 June 2011.


AC-dielectrophoretic characterization and separation of submicron and micron particles using sidewall AgPDMS electrodes.
The recent development of microfluidic "lab on a chip" devices requires the need to continuously separate submicron particles. Here, we present a PDMS microfluidic device with sidewall conducting PDMS (AgPDMS) composite electrodes capable of separating submicron particles in hydrodynamic flow. In particular, the device can service dual functions. First, the AgPDMS composite electrodes embedded in a sidewall of the device channel allow for performing AC-dielectrophoretic (DEP) characterization through direct microscopic observation of particle behavior. Characterization experiments are carried out for numerous parameters including particle size, medium conductivity, and AC field frequency to reveal important dielectrophoresis DEP information in terms of the crossover frequency and positive/negative DEP behavior under specific frequencies. Second, the device offers an advantage that sidewall AgPDMS composite electrodes can produce strong DEP effects throughout the entire channel height, and thus the robustness of the on-chip particle separation is demonstrated for continuous separation in a flowing mixture of 0.5 and 5 μm particles with 100% separation efficiency.


Biventricular support with the Jarvik 2000 axial flow pump: a feasibility study.
Patients with congestive heart failure who are supported with a left ventricular assist device (LVAD) may experience right ventricular dysfunction or failure that requires support with a right ventricular assist device (RVAD). To determine the feasibility of using a clinically available axial flow ventricular assist device as an RVAD, we implanted Jarvik 2000 pumps in the left ventricle and right atrium of two Corriente crossbred calves (approximately 100 kg each) by way of a left thoracotomy and then analyzed the hemodynamic effects in the mechanically fibrillated heart at various LVAD and RVAD speeds. Right atrial implantation of the device required no modification of either the device or the surgical technique used for left ventricular implantation. Satisfactory biventricular support was achieved during fibrillation as evidenced by an increase in mean aortic pressure from 34 mm Hg with the pumps off to 78 mm Hg with the pumps generating a flow rate of 4.8 L/min. These results indicate that the Jarvik 2000 pump, which can provide chronic circulatory support and can be powered by external batteries, is a feasible option for right ventricular support after LVAD implantation and is capable of completely supporting the circulation in patients with global heart failure.


Trichotillomania: a clinical study of 36 patients.
To understand the clinical characteristics of trichotillomania, 36 patients were investigated at the psychocutaneous special clinic from February 1982 to October 1990. There were 16 males and 20 females. The most prominent age was found in a group of 23 (63.9%) elementary school children, and in regards to the age of disease onset, 35 (97.2%) of our patients were under the age of 18. A 30-year-old woman with schizophrenia and a 6-year-old boy with mental retardation were noted in our study; the remaining 34 patients were generally well without signs of mental illness except for the recurrent failure to resist impulses to pull out their own hair. The common life events precipitating stress were academic problems and parent-child conflicts. The duration of follow-up was for at least 6 months. The habit of hair plucking was easily corrected if the duration of the disease was less than 6 months. Usually these patients had a satisfactory regrowth of hair within 1-3 months after our explanation and reassurance. In contrast, the patients with a hair loss of more than half a year were not easy to approach, and needed psychiatric treatment.


Ultrastructure of the aqueous lining layer in hamster airways: is there a two-phase system?
For particle retention and clearance, the structure and surface properties of the airway lining layer are important. Due to difficulties of its preservation, structural analysis has been hampered, and, hence, the existence of two distinct and continuous phases and how much osmiophilic material is available are unclear. It was the objective of this study to investigate the ultrastructure of the aqueous lining layer in the intrathoracic conducting airways of hamsters. By means of transmission electron microscopy, we investigated the ultrastructure of the airway lining layer in hamsters whose lungs have been fixed by the application of fixative dissolved in nonpolar fluorocarbon, either by instillation via the trachea or injection into the gas exchange parenchyma, together with intravascular perfusion of aqueous fixatives. The results were compared to lungs fixed by intravascular perfusion only. In twelve hamsters, the airway lining layer was found to consist of an aqueous phase and was coated by an osmiophilic film that follows fairly closely the upper-extending contours of cilia protruding from epithelial cells. Substantially less osmiophilic material was preserved in extrapulmonary airways and when nonaqueous fixative was injected. We found that the aqueous lining layer of the intrathoracic airways in hamsters essentially surrounds and covers the cilia, the microvilli, and any other structures like macrophages or deposited particles contained in it and is coated by an osmiophilic film of variable thickness. In healthy animals, a gel phase is expected to be very thin, not clearly separated from the periciliary fluid, and located just beneath the osmiophilic film.


Stop-bang questionnaire for screening obstructive sleep apnea syndrome among hypertensive patients in Kenya.
Obstructive sleep apnea (OSA) is a type of breathing problem during sleep caused by the blockage of the upper airway, which can cause cessation of airflow. There is limited research on the prevalence of OSA in hypertensive patients in sub-Saharan Africa (SSA). The study aimed to describe the prevalence and clinical characteristics of OSA among hypertensive patients at a tertiary hospital in Nairobi, Kenya. This cross-sectional study was conducted at the Aga Khan University Hospital in Nairobi, Kenya. Two hundred and fifty-one hypertensive patients were screened for OSA risk using the STOP-Bang questionnaire (SBQ). Patients with a SBQ score of ≥ 4 were categorized as high risk for OSA. Descriptive statistics were employed to describe both categorical and continuous variables and binary logistic regression to assess factors associated with the high risk of OSA. The study reported that 78.5% of the participants had high-risk OSA. The median age and body mass index (BMI) were 57.0 years (IQR: 50.0-64.0) and 28.3 kg/m2, respectively. Age, neck circumference, gender, and BMI were significantly higher in the high-risk OSA group as compared to the low-risk group. The study highlights the importance of screening hypertensive patients for OSA using the SBQ in clinical settings, particularly in low-and middle-income countries (LMICs). Healthcare providers can use patient characteristics such as age, gender, neck circumference, and BMI to identify those at greater risk of developing OSA. Further research could focus on developing effective OSA prevention and treatment interventions in hypertensive patients.


Sanfilippo disease in Greece.
In a series of cases collected in most parts of the world, Sanfilippo disease type A is more frequent than type B. Skin biopsies were obtained from Greek patients suspected for Sanfilippo disease and cultured fibroblasts were assayed for both N-acetyl-a-glucosaminidase and sulfamidase activity. Eleven patients with Sanfilippo disease were identified. Ten of them were type B and one type A. The 10 patients with type B came from East-Central Greece and the neighboring areas of Thessaly and Macedonia. Both parents of the type A patient were from the Greek ethnic community of Turkey. It remains unknown whether or not the higher frequency of type B than type A appears only in Greece or if it occurs in other Mediterranean countries as well.


Effects of nebracetam on synaptosomal monoamine uptake of striatal and hippocampal regions in rats.
Effects of nebracetam, a novel nootorpic agent, on the synaptosomal uptake of neurotransmitter monoamines of the brain regions were examined. Striatal and hippocampal synaptosomes were isolated by the Percoll gradient method, and the striatal dopamine uptake and hippocampal serotonin uptake were measured in the presence of different concentrations (1 to 1000 microM) of nebracetam in vitro. A significant reduction in dopamine uptake in the striatum and serotonin uptake in the hippocampus was seen at concentrations of 100 microM or above. In in vivo microdialysis study, there were no appreciable changes in the extracellular concentrations of striatal dopamine and hippocampal serotonin when this agent at a dose of 30 mg/kg, which was effective in improving ischemic brain energy metabolism, was applied i.p. to the rat. The ineffectiveness of nebracetam in the in vivo microdialysis may be due to low levels of the concentration of nebracetam when the agent was administered i.p. at a dose of 30 mg/kg, since the brain blood concentration of this agent is pharmacokinetically estimated to be no more than 15 microM when this dose of nebracetam is employed. Thus, it is unlikely that this agent at a pharmacologically effective dose alters dopamine or serotonin uptake in the brain nerve terminal under normal conditions.


Ectopic fat accumulation in the pancreas and its biomarkers: A systematic review and meta-analysis.
Presence of fat in the pancreas increases the risk of metabolic co-morbidities. Detection and quantification of pancreatic fat is not a routine clinical practice, at least in part because of need to use expensive imaging techniques. We aimed to systematically review common markers of pancreatic fat in blood and to investigate differences in these markers associated with fatty pancreas. The search was conducted in 3 databases (EMBASE, Scopus, and MEDLINE). Studies in humans were eligible for inclusion if they reported on biological markers and percentage of pancreatic fat or fatty pancreas prevalence. Data were pooled for correlation and effect size meta-analysis. A total of 17 studies including 11 967 individuals were eligible for meta-analysis. Markers of lipid metabolism, including circulating triglycerides (r = 0.38 [95% confidence interval (CI) 0.31, 0.46]) and high-density lipoprotein cholesterol (r = -0.33 [95% CI -0.35, -0.31]), and markers of glucose metabolism, including glycated haemoglobin (r = 0.39 [95% CI 0.30, 0.48], insulin (r = 0.38 [95% CI 0.33, 0.43]), and homeostasis model assessment-insulin resistance (r = 0.37 [95% CI 0.30, 0.44], yielded the best correlations with percentage of pancreatic fat. Further, effect size analysis showed large and medium effects for the above markers of lipid and glucose metabolism. Circulating levels of triglycerides and glycated haemoglobin appear to be the best currently available markers of pancreatic fat. The approach of non-invasive and accurate detection of pancreatic fat by blood analysis should be further explored in the future, by investigating other potential biological markers of pancreatic fat.


Haplotypes on pig chromosome 3 distinguish metabolically healthy from unhealthy obese individuals.
We have established a pig resource population specifically designed to elucidate the genetics involved in development of obesity and obesity related co-morbidities by crossing the obesity prone Göttingen Minipig breed with two lean production pig breeds. In this study we have performed genome wide association (GWA) to identify loci with effect on blood lipid levels. The most significantly associated single nucleotide polymorphisms (SNPs) were used for linkage disequilibrium (LD) and haplotype analyses. Three separate haploblocks which influence the ratio between high density lipoprotein cholesterol and total cholesterol (HDL-C/CT), triglycerides (TG) and low density lipoprotein cholesterol (LDL-C) levels respectively were identified on Sus Scrofa chromosome 3 (SSC3). Large additive genetic effects were found for the HDL-C/CT and LDL-C haplotypes. Haplotypes segregating from Göttingen Minipigs were shown to impose a positive effect on blood lipid levels. Thus, the genetic profile of the Göttingen Minipig breed seems to support a phenotype comparable to the metabolic healthy obese (MHO) phenotype in humans.


Susceptibility-weighted imaging findings of cortical laminar necrosis in pediatric patients.
MR susceptibility-weighted imaging (SWI) is a highly sensitive technique for detection of hemorrhage, but its utility in the evaluation of children with laminar necrosis is not yet known. We assessed whether cortical laminar necrosis in pediatric patients contains hemorrhage on SWI. "Cortical laminar necrosis" was defined as a hyperintense cortical lesion on T1-weighted imaging in the subacute or chronic phase of brain damage in some foci involving the cerebral cortex and white matter such as hypoxic-ischemic incidents and encephalopathy. Medical records, CT, and MR images were retrospectively analyzed. Fifteen patients (7 boys, 8 girls; age range, 0-13 years) were included. The areas of signal-intensity loss on SWI that were considered to be hemorrhage were correlated with the laminar necrosis. CT was assessed to correlate with the presence of calcification at the location of the signal-intensity loss on SWI. To assess appearance or signal-intensity changes of hemorrhage in the laminar necrosis, follow-up SWI was performed. The causes of laminar necrosis included infarction in 4 patients, ischemic changes from Moyamoya disease in 2, meningoencephalitis in 2, hypoxic-ischemic encephalopathy in 2, shaken baby syndrome in 1, encephalopathy from severe infection in 1, status epilepticus in 1, citrullinemia in 1, and brain injury with posterior reversible encephalopathy syndrome in 1. T1-weighted imaging showed focal laminar necrosis in 8, multifocal laminar necrosis in 2, and diffuse laminar necrosis in 5. SWI findings correlated with laminar necrosis included the following: no hemorrhage in 13 patients (80.0%), dotted hemorrhage in 2 (13.3%), and laminar hemorrhage in 1 (6.7%). Follow-up SWI performed in 6 patients showed no additional hemorrhage. Most areas of cortical laminar necrosis in pediatric patients showed no hemorrhage on SWI.


Neuronal transgene expression in dominant-negative SNARE mice.
Experimental advances in the study of neuroglia signaling have been greatly accelerated by the generation of transgenic mouse models. In particular, an elegant manipulation that interferes with astrocyte vesicular release of gliotransmitters via overexpression of a dominant-negative domain of vesicular SNARE (dnSNARE) has led to documented astrocytic involvement in processes that were traditionally considered strictly neuronal, including the sleep-wake cycle, LTP, cognition, cortical slow waves, depression, and pain. A key premise leading to these conclusions was that expression of the dnSNARE was specific to astrocytes. Inconsistent with this premise, we report here widespread expression of the dnSNARE transgene in cortical neurons. We further demonstrate that the activity of cortical neurons is reversibly suppressed in dnSNARE mice. These findings highlight the need for independent validation of astrocytic functions identified in dnSNARE mice and thus question critical evidence that astrocytes contribute to neurotransmission through SNARE-dependent vesicular release of gliotransmitters.


[Progress in finite element analysis of meniscus].
The meniscus biomechanics is one of the hottest areas in medical research. Finite element analysis(FEA) provides a new method for biomechanical studies. Using the FEA, the stress force and strain distribution characteristics of the normal meniscus can be analysed by establishing the finite element model of the meniscus and utilizing finite element softwares. In addition, the finite simulation of the meniscus tears, meniscectomy, and knee tissue injury can assess the changes of meniscus biomechanical properties. These research results provide theoretical basis of biomechanics to prevent and treat knee joint injuries. However, the finite element analysis method has its limitations, future research will be aimed at establishing high quality finite element models, making the models offer more authentic simulations of the meniscus anatomy, and ensuring the FEA could be applied to research a variety of knee injuries.


Schizophrenia risk and paternal age: a potential role for de novo mutations in schizophrenia vulnerability genes.
How schizophrenia (SZ) is maintained at roughly 1% of the population despite diminished reproduction is one puzzle currently facing researchers. De novo mutations were first proposed over half a century ago as a source for new SZ genes. Current evidence linking advancing paternal age to SZ risk makes revisiting this hypothesis important. Advancing paternal age is the major source of new mutations in the human population. This article will examine potential mechanisms whereby parental age may impact new mutations, as well as review recent data supporting such a hypothesis.


Pollutant-induced modulation in conformation and β-lactamase activity of human serum albumin.
Structural changes in human serum albumin (HSA) induced by the pollutants 1-naphthol, 2-naphthol and 8-quinolinol were analyzed by circular dichroism, fluorescence spectroscopy and dynamic light scattering. The alteration in protein conformational stability was determined by helical content induction (from 55 to 75%) upon protein-pollutant interactions. Domain plasticity is responsible for the temperature-mediated unfolding of HSA. These findings were compared to HSA-hydrolase activity. We found that though HSA is a monomeric protein, it shows heterotropic allostericity for β-lactamase activity in the presence of pollutants, which act as K- and V-type non-essential activators. Pollutants cause conformational changes and catalytic modifications of the protein (increase in β-lactamase activity from 100 to 200%). HSA-pollutant interactions mediate other protein-ligand interactions, such as HSA-nitrocefin. Therefore, this protein can exist in different conformations with different catalytic properties depending on activator binding. This is the first report to demonstrate the catalytic allostericity of HSA through a mechanistic approach. We also show a correlation with non-microbial drug resistance as HSA is capable of self-hydrolysis of β-lactam drugs, which is further potentiated by pollutants due to conformational changes in HSA.


Characterization of carbon nanofibre-reinforced polypropylene foams.
In this paper, carbon-nanofibre-reinforced polypropylene foams were prepared and characterized regarding their foaming behaviour, cellular structure and both thermo-mechanical as well as electrical properties. Polypropylene (PP) nanocomposites containing 5, 10 and 20 wt% of carbon nanofibres (CNF) and a chemical blowing agent were prepared by melt-mixing inside a twin-screw extruder and subsequently water-cooled and pelletized. The extruded nanocomposites were later foamed using a one-step compression-moulding process. The thermo-mechanical properties of the CNF-reinforced PP foams were studied, analyzing the influence of the carbon nanofibres on the cellular structure and subsequent thermo-mechanical behaviour of the foams. Carbon nanofibres not only seemed to act as nucleating agents, reducing the average cell size of the foams and increasing their cell density for similar expansion ratios, but also helped produce mechanically-improved foams, even reaching for the 20 wt% CNF-reinforced ones a specific modulus around 1.2 GPa x cm3/g for densities as low as 300 kg/m3. An increasingly higher electrical conductivity was assessed for both the solids as well as the foams with increasing the amount of carbon nanofibres.