The frequency and influence of gallbladder varices on gallbladder functions in patients with portal hypertension.
Gallbladder varices have been reported in patients with portal hypertension. The exact frequency and significance of these collaterals in patients with cirrhotic and noncirrhotic portal hypertension is not known. One hundred and two patients with portal hypertension [38 with cirrhosis, 29 with noncirrhotic portal fibrosis (NCPF) and 35 with extrahepatic portal vein obstruction (EHPVO)] and 25 healthy controls were studied. Gallbladder varices were seen at ultrasound as tortuous, dilated vessels in the wall or in the bed of the gallbladder. In 35 patients (19 patients with and 16 without gallbladder varices) and in 10 healthy controls, gallbladder functions were studied by determining fasting volume (FV) and then residual volume (RV) every 10 min over 1 h after giving a liquid meal of 420 k.cal. Ejection fraction (EF) was computed as a percentage by the formula: FV--RV/FV x 100. Twenty four (24%) patients had gallbladder varices: Five (13%) with cirrhosis, seven (24%) with NCPF, and 12 (34%) with EHPVO. FV in EHPVO patients was seen significantly more than in cirrhotics (31.6 +/- 15.4 vs 19.3 +/- 6.0 ml, p < 0.05). The RV and EF were not different in the three groups of patients compared with the controls. The EF was similar in patients with or without gallbladder varices (63.3 +/- 10.2% vs 64.6 +/- 10.4%). Gallbladder varices are often seen in portal hypertension, more often in EHPVO patients, and these collaterals cause some gallbladder stasis but do not impede gallbladder function and hence seem unlikely to contribute to gallstone formation.


Identification of the rate of chimerism of different tissues with microsatellite markers in chicken chimeras.
The goal of our study was to evaluate whether private alleles can be defined in microsatellite markers for the breeds under investigation; to evaluate if these private alleles distinguish chicken chimera when using different tissues; to trace them back to the donor: Green-Legged Partridgelike and recipient: White Leghorn chicken breeds, and further on, to estimate the level of chimerism in each tissue. Private and common alleles were defined for donor and recipient chicken breeds in 3 loci. The rate of chimerism was defined based on private alleles present in liver, heart, breast muscle, femoral muscle and gonads. The highest rate of chimerism was observed in liver. A lower rate of chimersim was observed in gonads, and femoral muscle, and finally the lowest rate of chimerism was observed in breast muscle and heart.


[Immunohistochemical study of lactate dehydrogenase isoenzymes in gastric cancer: light and electron microscopic observations].
In order to study the metabolic features of gastric cancer, the relationship between intestinal metaplasia or dysplasia and gastric cancer, and the relationship between LDH isoenzymes and biological behavior of gastric cancer, the content and distribution of lactate dehydrogenase (LDH) isoenzymes in 60 cases of gastric cancer were observed with immunohistochemical and immunoelectron microscopic techniques. The results showed that the content of LDH5 in gastric cancer cells was higher than that in any cells of normal gastric epithelium except parietal cells (P < 0.001). LDH-H (LDH1-LDH4) in cancer cells was lower than in parietal cells, but was not significantly different from that in chief cells and surface epithelium. Under electron microscope, LDH5 in cancer cells was mainly distributed in the matrix of cytoplasm, while LDH-H was scarce. LDH5 in the epithelium of intestinal metaplasia (IM) or dysplasia was higher than in pyloric glandular epithelium (P < 0.05), but was not significantly different from that in cancer cells (P > 0.2). LDH-H in the epithelium of IM or dysplasia was higher than that in pyloric glandular epithelium and gastric cancer cells (P < 0.05). The results suggest that increased LDH in gastric cancer cells is resulted mainly from increased LDH5, which makes the gastric cancer cells produce more lactate, with decrease in local pH. Invasion and spread of cancer cells could be indirectly promoted through an elevated activity of acid hydrolases. IM and dysplasia may be considered as borderline lesions from normal gastric mucosa to gastric cancer.


Diabetes Care in Iran: Where We Stand and Where We Are Headed.
The prevalence of diabetes has steadily increased in Iran from the time of the first published nationally representative survey in 1999 and despite efforts and strategies to reduce disease burden. The aim of the present review was to describe the current status of diabetes care in Iran. A selective review of the relevant literature, focusing on properly conducted studies, describing past and present diabetes care strategies, policies, and outcomes in Iran was performed. The quality of diabetes care has gradually improved as suggested by a reduction in the proportion of undiagnosed patients and an increase in affordability of diabetes medications. The National Program for Prevention and Control of Diabetes has proven successful at identifying high-risk individuals, particularly in rural and remote-access areas. Unfortunately, the rising tide of diabetes is outpacing these efforts by a considerable margin. Substantial opportunities and challenges in the areas of prevention, diagnosis, and management of diabetes exist in Iran that need to be addressed to further improve the quality of care and clinical outcomes.


Reversal of age-related effects in rat muscle phosphoglycerate kinase.
Rat muscle phosphoglycerate kinase is one of several enzymes in which age-related effects have been identified. Thus, samples of this enzyme isolated from old rats display a greatly increased heat stability as compared with enzyme isolated from young animals. Previous studies detected no differences in the sequence of amino acids or in the net charge between the young and old forms of the enzyme and it was concluded that the age-related structural modifications are purely conformational. The present study was conducted with the aim of critically testing this hypothesis. To this end, samples of phosphoglycerate kinase purified from skeletal muscle of young and old rats were unfolded by an 18-hr incubation in a 2 M guanidine hydrochloride solution at 4 degrees C, a treatment that results in extensive loss of the three-dimensional structure of the enzyme. A complete reactivation of both enzymes was achieved by dilution of the unfolded enzyme solutions into a large excess of denaturant-free buffer followed by 4 hr of incubation at 25 degrees C. The reactivation kinetics of the unfolded young and old enzymes were practically identical and the refolded products, compared using heat-inactivation kinetics as a sensitive probe, were found to be identical. Moreover, their heat inactivation coincided with that of young untreated phosphoglycerate kinase. These results demonstrate the reversibility of age-related effects at the molecular level and provide strong support for the hypothesis that the modifications in phosphoglycerate kinase in old muscle are purely conformational and, hence, clearly postsynthetic.


Immeasurable levels of serum phosphate--an unidentified cause of respiratory failure in a diabetic patient.
We report a case of immeasurable levels of serum phosphate in a patient with juvenile type Diabetes mellitus and diabetic ketoacidosis who developed respiratory failure. A 27-year-old female with juvenile type insulin-dependent Diabetes mellitus was admitted because of suspected acute mediastinitis and respiratory failure, probably, among other responsible factors, caused and complicated by undetectable levels of serum phosphate. The serum phosphate concentration three days after aggressive treatment was only 0.2 mmol/L. Furthermore, a significant improvement in weakness and lethargy was observed. To the best of our knowledge, this is the first described case of immeasurable levels of serum phosphate. In patients with Diabetes mellitus, serum phosphate concentrations should be routinely checked in order to avoid additional complications.


Reversible hypothyroidism and Whipple's disease.
The major cause of primary hypothyroidism is autoimmune mediated with progressive and permanent destruction of the thyroid gland resulting in life-long replacement therapy. Treatable and reversible hypothyroidism is unusual and here forth is such a case due to infection of the thyroid gland with Tropheryma whippleii, Whipple disease. A 45 year-old female presented with symptoms and signs consistent with primary hypothyroidism, which was also confirmed biochemically. Her response to thyroxine replacement therapy was poor however, requiring a significantly elevated amount. Further investigation revealed the presence of Whipple's disease involving the gastrointestinal trace and possibly the thyroid gland. Her thyroxine requirement decreased drastically following appropriate antimicrobial therapy for Whipple's disease to the extent that it was ceased. Thyrotropin releasing hormone testing in the steady state suggested there was diminished thyroid reserve due to Whipple's disease. This is the first ante-mortem case report studying the possible involvement of the thyroid gland by Whipple's disease. Despite the normalization of her thyroid function test biochemically after antibiotic therapy, there is diminished thyroid reserve thus requiring close and regular monitoring.


Geometric fractal growth model for scale-free networks.
We introduce a deterministic model for scale-free networks, whose degree distribution follows a power law with the exponent gamma. At each time step, each vertex generates its offspring, whose number is proportional to the degree of that vertex with proportionality constant m-1 (m>1). We consider the two cases: First, each offspring is connected to its parent vertex only, forming a tree structure. Second, it is connected to both its parent and grandparent vertices, forming a loop structure. We find that both models exhibit power-law behaviors in their degree distributions with the exponent gamma = 1+ln(2m-1)/ln m. Thus, by tuning m, the degree exponent can be adjusted in the range, 2 < gamma < 3. We also solve analytically a mean shortest-path distance d between two vertices for the tree structure, showing the small-world behavior, that is, d approximately ln N/ln K macro, where N is system size, and k macro is the mean degree. Finally, we consider the case that the number of offspring is the same for all vertices, and find that the degree distribution exhibits an exponential-decay behavior.


Involvement of the membrane distal catalytic domain in pervanadate-induced tyrosine phosphorylation of receptor protein-tyrosine phosphatase alpha.
Receptor protein-tyrosine phosphatase alpha, RPTPalpha, is a typical transmembrane protein-tyrosine phosphatase (PTP) with two cytoplasmic catalytic domains. RPTPalpha became strongly phosphorylated on tyrosine upon treatment of cells with the PTP inhibitor pervanadate. Surprisingly, mutation of the catalytic site Cys in the membrane distal PTP domain (D2), but not of the membrane proximal PTP domain (D1) that harbors the majority of the PTP activity, almost completely abolished pervanadate-induced tyrosine phosphorylation. Pervanadate-induced RPTPalpha tyrosine phosphorylation was not restricted to Tyr789, a known phosphorylation site. Cotransfection of wild-type RPTPalpha did not potentiate tyrosine phosphorylation of inactive RPTPalpha-C433SC723S, suggesting that RPTPalpha-mediated activation of kinase(s) does not underlie the observed effects. Mapping experiments indicated that pervanadate-induced tyrosine phosphorylation sites localized predominantly, but not exclusively, to the C-terminus. Our results demonstrate that RPTPalpha-D2 played a role in pervanadate-induced tyrosine phosphorylation of RPTPalpha, which may suggest that RPTPalpha-D2 is involved in protein-protein interactions.


Down-regulation of liver drug-metabolizing enzymes in a murine model of chronic renal failure.
Drug metabolism could be altered in patients with chronic renal failure (CRF). In rats, this phenomenon is related to a decrease in liver cytochrome P450 (P450) and phase II enzymes, particularly N-acetyltransferase 2 (NAT2). This study attempted to determine the effects of CRF on liver P450 isoforms and NAT2 expressions by using a CRF mouse model. Two groups of mice were studied: CRF induced by 3/4 nephrectomy and control. Liver protein expression and mRNA levels of the major P450 isoforms involved in drug metabolism (CYP1A2, 2C29, 2D, 2E1, and 3A11) and NAT2 were measured by Western blot and real-time polymerase chain reaction (PCR), respectively. CYP3A activity was also assessed by the N-demethylation of erythromycin. Results showed a significant reduction in the protein expression of CYP1A2 (56%), 2C29 (31%), and 3A11 (37%) in CRF mice compared with control animals. Real-time PCR revealed a similar reduction in mRNA levels of CYP1A2, 2C29, and 3A11 (59, 56, and 37%, respectively), in CRF mice. There was no significant modification in protein expression and mRNA of CYP2D and 2E1. Compared with control animals, CRF mice displayed a 25% reduction in N-demethylation of erythromycin. For NAT2, protein expression decreased by 33% and mRNA levels decreased by 23%. In conclusion, this study demonstrates that protein expression of liver CYP1A2, CYP2C29, and CYP3A11 is down-regulated in CRF mice, secondary to reduced gene expression. Phase II enzymes are similarly affected by CRF. Our results will allow the use of knockout mice to determine the mechanism underlying CRF-induced down-regulation of liver drug-metabolizing enzymes.


Reversible circularization of an anthracene-modified DNA conjugate through bimolecular triplex formation and its analytical application.
We prepared an oligodeoxyribonucleotide conjugate (5-3ant(2)18) carrying two anthracenes, each of which was tethered to both ends of the conjugate through hexamethylene linker chains. The conjugate has a mirror repeat of two heptamer sequences, such that it forms a bimolecular triplex with the single stranded target, forming a two-fold U-shaped conformation. The conformation of the conjugate in its triplex structure could be frozen instantaneously by circularization through photodimerization of the anthracenes. Compared with the duplex formation of linear probes with relevant sequences, bimolecular triplex formation of 5-3ant(2)18 shows a unique feature in its target recognition; it binds the target tightly, yet still retains high sequence selectivity. Circularization of 5-3ant(2)18 by UV photoirradiation was verified as the probe reaction for a DNA assay. The probe reaction could be performed in a few seconds over a wide range of temperatures, at least between 0 and 25 °C. In addition, the reaction could be regarded as a reversible method for the preparation of circular DNA that shows higher affinity for the target.


Simultaneous determination of cardenolides by sonic spray ionization liquid chromatography-ion trap mass spectrometry--a fatal case of oleander poisoning.
Simultaneous determination of oleandrin and its three related compounds, desacetyloleandrin, oleandrigenin, and gitoxigenin in blood by using liquid chromatography-three-dimensional quadrupole mass spectrometry (LC-3DQMS) system equipped with sonic spray ionization (SSI) interface was conducted. This analyzing method was suitable for all of these compounds except gitoxigenin. The limits of detection of oleandrigenin and desacetyloleandrin from blood were 2 ng/mL and that of oleandrin was 3 ng/mL. The calibration curves for oleandrin, desacetyloleandrin, and oleandrigenin were linear in the range of 5-100 ng/mL. The coefficients of variation of oleandrin, desacetyloleandrin, and oleandrigenin in the blood were satisfactory ranging from 1.6% to 4.1%. This analysis method was applied to a fatal case of oleander poisoning. As a result of liquid chromatography-mass spectrometry (LC-MS) analysis, oleandrin was detected in heart blood and cerebrospinal fluid. Desacetyloleandrin, oleandrigenin, and gitoxigenin were not detected. In order to make identification of oleandrin reliable, LC-MS-MS analysis was performed. The concentrations of oleandrin found in the heart blood and cerebrospinal fluid were 9.8 and 10.1 ng/mL, respectively.


Electron-positron pair production in the field of superstrong oppositely directed laser beams.
The multiphoton electron-positron pair production through nonlinear channels by superintense opposed laser pulses of the same frequencies in vacuum is considered. On the basis of the Dirac model the resonance approximation for vacuum induced transitions is developed. The analytic formulas for energetic and angular distributions and total number of created particles in the limit of a short interaction time are obtained.


Additive effect on reductive decomposition and binding of carbonate-based solvent toward solid electrolyte interphase formation in lithium-ion battery.
The solid-electrolyte interphase (SEI) formed through the reductive decomposition of solvent molecules plays a crucial role in the stability and capability of a lithium-ion battery (LIB). Here we investigated the effects of adding vinylene carbonate (VC) to ethylene carbonate (EC) solvent, a typical electrolyte in LIBs, on the reductive decomposition. We focused on both thermodynamics and kinetics of the possible processes and used density functional theory-based molecular dynamics with explicit solvent and Blue-moon ensemble technique for the free energy change. We considered Li(+) in only EC solvent (EC system) and in EC solvent with a VC additive (EC/VC system) to elucidate the additive effects. In addition to clarifying the equilibrium properties, we evaluated the free energy changes along several EC or VC decomposition pathways under one-electron (1e) reduction condition. Two-electron (2e) reduction and attacks of anion radicals to intact molecules were also examined. The present results completely reproduce the gaseous products observed in the experiments. We also found a new mechanism involving the VC additive: the VC additive preferentially reacts with the EC anion radical to suppress the 2e reduction of EC and enhance the initial SEI formation, contrary to the conventional scenario in which VC additive is sacrificially reduced and its radical oligomerization becomes the source of SEI. Because our mechanism needs only 1e reduction, the irreversible capacity at the SEI formation will decrease, which is also consistent with the experimental observations. These results reveal the primary role of VC additive in the EC solvent.


A mortality study on male subjects exposed to polyfluoroalkyl acids with high internal dose of perfluorooctanoic acid.
The aim of the present study was to examine the association between exposure to polyfluoroalkyl substances (PFASs) and mortality (1970-2018) in a cohort of 462 male employees who had worked at least six months before 2009 for a factory (14,658 person-years; 107 deaths, average follow-up time 31.7 years), which had been producing perfluorooctanoic acid (PFOA), perfluorooctanesulfonyl fluoride (PFOS) and other chemicals since 1968. Employees were classified as follows: 1) by probability of exposure to PFASs; 2) by tertiles of PFOA serum concentrations. In a fraction (n = 120) of workers measurements of internal PFOA serum concentration were used to predict a cumulative serum PFOA concentration of each cohort member. Mortality rates were compared to that of the regional population using the standardized mortality ratio (SMR), and to that of the workers of a nearby metalworking factory in terms of risk ratio (RR), across categories of probability of PFASs exposure and tertiles of cumulative serum PFOA concentrations. Internal PFOA serum concentration among 120 workers in the 2000-2013 period was very high (Geometric Mean: 4048 ng/mL; range 19-91,900 ng/mL). The mortality of the chemical cohort was increased for liver cancer (SMR: 2.32; CI: 1.11-4.87), malignant neoplasm of lymphatic and haematopoietic tissue (SMR: 2.26; CI: 1.08-4.73). In the comparison with the cohort of workers from the metalworking factory, the RRs for mortality of the cohort were increased for overall mortality (RR: 1.42; CI: 1.12-1.79), diabetes (RR: 5.95; CI: 1.08-32.8), liver cancer (RR: 6.69; CI: 1.71-26.2) and liver cirrhosis (RR: 3.87; CI: 1.18-12.7). Mortality for these causes increased in association with probability of PFASs exposure and with tertiles of cumulative PFOA serum concentrations. The present is a small observational study with limited control over confounding factors. The cohort showed increased mortality for all causes and subjects in the highest cumulative internal dose of PFOA had a statistically significant increase for mortality of liver cancer, liver cirrhosis, diabetes, malignant neoplasms of lymphatic and haematopoietic tissue in both comparisons. Toxicological studies on PFOA and PFOS provide support for causality for the observed association with the risk for liver cirrhosis and liver cancer.


Assessment of preference for breast cancer chemoprevention in Japanese young women.
Pills containing estrogen and progesterone or gonadotropin releasing hormone agonist have been considered valuable to prevent breast cancer. This study assessed preference for the combination-type pill for preventing breast cancer, to evaluate the hypothetical preventive effect of this agent among young Japanese women. The standard gamble method was applied. Fifty-five college students and 44 nursing school students aged between 18 and 41 years were asked to decide the probability of being affected by breast cancer at which they would start to take this agent. Preference score was calculated by subtracting the probability given by each respondent from 1, which corresponds to the value (utility) she allotted to the agent. The means of preference score were 0.58, 0.48, 0.37, and 0.27 for 100, 75, 50, and 25% of efficacy levels of the agent, respectively. Preference score was significantly lower in nursing school students and those whose knowledge about hormones were relatively high. Score of Health Locus of Control (HLC) was nonsignificantly negatively correlated with preference score at any efficacy level. HLC score was significantly higher among those who refused the agent with 50 and 25% efficacy levels at 100% level of breast cancer risk. The data suggest that perceived risk of this agent was not negligibly small in this population and school status, knowledge about hormones, and beliefs about health would affect preference for the agent. Understanding of preference for chemopreventive agents for breast cancer, especially those containing hormones, is important to assess their potential as future preventive agents and is helpful when planning a strategy of chemoprevention.


Anchor-wire technique for multiple plastic biliary stents to prevent stent dislocation.
In endoscopic placement of multiple plastic biliary stents (PBSs), we sometimes experience proximal dislocation of the first PBS at the time of subsequent PBS insertion. We describe the case of a 79-year-old male with obstructive jaundice caused by cholangiocarcinoma who needed to receive multiple PBS placements for management of cholangitis. Although proximal dislocation of the first PBS was observed, we prevented the dislocation via our technique of using guidewire inserted from the distal end of the first PBS to the side hole as the anchor-wire. We could complete this technique only by inserting guidewire through the side hole of the first PBS during the process of releasing the first PBS and pulling out the guidewire and the inner sheath. It did not matter whether the anchor-wire went towards the third portion of the duodenum or the duodenal bulb. Here we introduce this "anchor-wire technique", which is useful for the prevention of PBS proximal dislocation in placing multiple PBSs.


Fostering teaching-learning through workplace based assessment in postgraduate chemical pathology residency program using virtual learning environment.
The principle of workplace based assessment (WBA) is to assess trainees at work with feedback integrated into the program simultaneously. A student driven WBA model was introduced and perception evaluation of this teaching method was done subsequently by taking feedback from the faculty as well as the postgraduate trainees (PGs) of a residency program. Descriptive multimethod study was conducted. A WBA program was designed for PGs in Chemical Pathology on Moodle and forms utilized were case-based discussion (CBD), direct observation of practical skills (DOPS) and evaluation of clinical events (ECE). Consented assessors and PGs were trained on WBA through a workshop. Pretest and posttest to assess PGs knowledge before and after WBA were conducted. Every time a WBA form was filled, perception of PGs and assessors towards WBA, time taken to conduct single WBA and feedback were recorded. Faculty and PGs qualitative feedback on perception of WBA was taken via interviews. WBA tools data and qualitative feedback were used to evaluate the acceptability and feasibility of the new tools. Six eligible PGs and seventeen assessors participated in this study. A total of 79 CBDs (assessors n = 7 and PGs n = 6), 12 ECEs (assessors n = 6 and PGs n = 5), and 20 DOPS (assessors n = 6 and PGs n = 6) were documented. PGs average pretest score was 55.6%, which was improved to 96.4% in posttest; p value< 0.05. Scores of annual assessment before and after implementation of WBA also showed significant improvement, p value 0.039, Overall mean time taken to evaluate PG's was 12.6 ± 9.9 min and feedback time 9.2 ± 7.4 min. Mean WBA process satisfaction of assessors and PGs on Likert scale of 1 to 10 was 8 ± 1 and 8.3 ± 0.8 respectively. Both assessors and fellows were satisfied with introduction and implementation of WBA. It gave the fellows opportunity to interact with assessors more often and learn from their rich experience. Gain in knowledge of PGs was identified from the statistically significant improvement in PGs' assessment scores after WBA implementation.


Diosgenin ameliorates palmitic acid-induced lipid accumulation via AMPK/ACC/CPT-1A and SREBP-1c/FAS signaling pathways in LO2 cells.
Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and is characterized by excessive hepatic lipid accumulation. Many studies have suggested that lipid overload is the key initial factor that contributes to hepatic steatosis. Our previous study indicated that diosgenin (DSG) has a beneficial effect on energy metabolism, but the underlying mechanism remains unclear. Human normal hepatocytes (LO2 cells) were incubated with palmitic acid to establish the cell model of nonalcoholic fatty liver. The effects of DSG on lipid metabolism, glucose uptake and mitochondrial function were evaluated. Furthermore, the mechanism of DSG on oxidative stress, lipid consumption and lipid synthesis in LO2 cells was investigated. The results indicated that palmitic acid induced obvious lipid accumulation in LO2 cells and that DSG treatment significantly reduced the intracellular lipid content. DSG treatment upregulated expression of lipolysis proteins, including phospho-AMP activated protein kinase (p-AMPK), phospho-acetyl-coA carboxylase (p-ACC) and carnitine acyl transferase 1A (CPT-1A), and inhibited expression of lipid synthesis-related proteins, including sterol regulatory element-binding protein 1c (SREBP-1c) and fatty acid synthase (FAS). Additionally, DSG-treated cells displayed a marked improvement in mitochondrial function, with less production of reactive oxygen species and a higher mitochondrial membrane potential compared with the model group. This study suggests that DSG can reduce intracellular lipid accumulation in LO2 cells and that the underlying mechanism may be related to the improving oxidative stress, increasing fatty acid β-oxidation and decreasing lipid synthesis. The above changes might be mediated by the activation of the AMPK/ACC/CPT-1A pathway and inhibition of the SREBP-1c/FAS pathway.


Study of caspase inhibitors for limiting death in mammalian cell culture.
Apoptosis in mammalian cell culture is associated with decreased bioproduct yields and can be inhibited through altering the intracellular signaling pathways mediating programmed cell death. In this study, we evaluated the capacity to inhibit caspases to maintain high viable cell numbers in CHO and 293 cultures. Two genetic caspase inhibitors, XIAP and CrmA, were examined along with a mutant of each, XIAP-BIR123NC, which contains three BIR domains but lacks the RING finger, and CrmA-DQMD, which has CrmA's pseudosubstrate site replaced with that of another caspase inhibitor, p35. Stable CHO pooled and 293 clonal cell lines expressing each protein were exposed to apoptotic insults, including spent medium, Sindbis virus, and etoposide. For each insult the mutated protein resulted in higher viabilities than its wild-type counterpart. However, the mutants provided different levels of protection, depending on the insult considered. CrmA-DQMD was the preferred inhibitor for spent medium-induced apoptosis, whereas XIAP-BIR123NC conferred better protection for etoposide-induced death. Addition of Z-VAD.fmk to the genetically engineered cells enhanced viabilities in the presence of spent medium or etoposide; however, the largest increases in viability were experienced by the control cells, indicating an overlap in caspase inhibition between the genetic and chemical inhibitors. Finally, parental 293 cells were treated with caspase-8 and -9 inhibitors, Z-IETD.fmk and Z-LEHD.fmk, in concert with spent medium or etoposide exposure. Spent medium-induced death was delayed more readily with the caspase-8 inhibitors, CrmA-DQMD and Z-IETD.fmk, and etoposide-induced death was stalled more so with XIAP-BIR123NC and Z-LEHD.fmk. These results suggest that the apoptosis pathways induced and the level of protection afforded by a particular caspase inhibitor may vary with the insult considered.


Crisis standards of care in a pandemic: navigating the ethical, clinical, psychological and policy-making maelstrom.
The COVID-19 pandemic has caused clinicians at the frontlines to confront difficult decisions regarding resource allocation, treatment options and ultimately the life-saving measures that must be taken at the point of care. This article addresses the importance of enacting crisis standards of care (CSC) as a policy mechanism to facilitate the shift to population-based medicine. In times of emergencies and crises such as this pandemic, the enactment of CSC enables concrete decisions to be made by governments relating to supply chains, resource allocation and provision of care to maximize societal benefit. This shift from an individual to a population-based societal focus has profound consequences on how clinical decisions are made at the point of care. Failing to enact CSC may have psychological impacts for healthcare providers particularly related to moral distress, through an inability to fully enact individual beliefs (individually focused clinical decisions) which form their moral compass.


A novel rapid and accurate method for detecting Helicobacter Pylori: the modified antigen test.
We aim to assess the diagnostic accuracy of the gastric biopsy Helicobacter pylori (H. pylori) antigen stool test as a novel method for detecting H. pylori, comparing this test with the histopathological evaluation of H. pylori and H. pylori antigen stool test as the reference standards. The study involves patients who are scheduled for an upper gastrointestinal endoscopy. Gastric biopsies were endoscopically obtained from all patients, and H. pylori antigen stool tests were performed for all patients. Results from the gastric biopsies that were studied using the H. pylori antigen stool test in terms of the novel method were obtained and recorded. The inter-rater agreement between the H. pylori tests in determining positive and negative results was investigated using Fleiss' and Cohen's kappa tests. The capacity of applied tests in predicting the presence of H. pylori was analyzed using a receiver operating characteristic (ROC) curve analysis. A total of 55 patients were studied (32 females and 23 males). The strongest coherence was obtained between the gastric biopsy test and histopathological evaluation with a kappa value of 0.664 in Cohen's kappa analysis of overall coherence between tests. The most accurate sensitivity and specificity values were obtained for the gastric biopsy test and histopathological evaluation crosstabulation for both overall comparisons at 90.5% sensitivity and 79.4% specificity. With this new, rapid, and easy-to-apply method, patients' endoscopies and gastric biopsies looking for the presence of H. pylori would be determined with more sensitive and more specific accuracy rates than current antigen stool tests, and H. pylori can be eradicated immediately without waiting for the histopathological evaluation period.


SH2 domain function is essential for the role of the Lck tyrosine kinase in T cell receptor signal transduction.
Tyrosine kinase activity is required for signal transduction through the T cell antigen receptor (TCR). The Src family tyrosine kinase Lck appears to play a key role in the initiation of TCR signaling events. We have investigated the role of the phosphotyrosine-binding Src homology-2 (SH2), domain of Lck in TCR signaling. Lck containing a mutation in the phosphotyrosine binding pocket of the SH2 domain was expressed in an Lck-deficient cell line. We found that, in contrast to wild-type Lck, the SH2 domain mutant was unable to restore even the earliest TCR-mediated signaling events. To investigate the role of the Lck SH2 domain, we examined the association of tyrosine phosphoproteins with Lck. The predominant associated phosphoprotein was the ZAP-70 tyrosine kinase, which has also been implicated in the initiation of TCR signaling. In addition, the zeta subunit of the T cell receptor was found to weakly associate with Lck. Further analysis indicated that the SH2 domain of Lck can directly recognize both ZAP-70 and zeta in immunoprecipitates from TCR-stimulated cells. Our findings demonstrate that the SH2 domain of Lck is essential for the initiation of signaling events following TCR stimulation probably as a result of its ability to mediate an interaction between Lck and the ZAP-70 tyrosine kinase and/or the zeta subunit of the T cell receptor.


Investigating of labelling and detection of transthyretin synthetic peptide derivatized with naphthalene-2,3-dicarboxaldehyde.
Labelling and detection of a synthetic peptide (PN) mimicking a tryptic fragment of interest for the diagnosis of familial amyloidal polyneuropathy have been investigated optically and electrochemically. We decided to covalently label naphtalene-2,3-dicarboxyaldehyde (NDA), a fluorogenic and electroactive molecule on PN. First, the optimization of the labelling chemical reaction was performed by capillary electrophoresis coupled with laser induced fluorescence detection (CE-LIF). The analytical parameters such as separation efficiency and peak area were considered to propose this optimized derivatization reaction. The results obtained allowed us to establish the pH and ionic strength of the derivatization buffer, the molar ratio between NDA and PN and the reaction time of the labelling. Optimal conditions are obtained when [NDA]/[PN]=40, buffer pH of 9, buffer ionic strength of 70 mM and reaction time of 15 min. Second, differential pulse voltammetry (DPV) and cyclic voltammetry (CV) were also used to characterize NDA-labelled PN and different electroinactive amino acids (histidine, lysine, serine, threonine) which are in the PN sequence. The electrochemical detection experiments demonstrated that the labelled biomolecules could be also easily detected at low concentration. Moreover, the derivatization reaction could be followed to describe more precisely the labelling process of these biomolecules. Optimal conditions for labelling are obtained when [NDA]total/[CN(-)] ratio =1 and [NDA]total/[amino acid or peptide]=100 with a buffer having a pH=9 on a glassy carbon electrode. In all cases, an obvious oxidation peak for the N-2-substituted-1-cyanobenz-[f]-isoindole derivative (CBI) has been observed at 0.5-0.7 V/SCE. The multi-labelling of PN and lysine were shown with DPV. We presumed this result to occur because of the shouldered shape of the DPV peak shape. These experiments confirm that NDA can be used as a derivative agent for PN, allowing for electrochemical and fluorescence detections with a limit of detection of labelled PN estimated at 0.2 µM and 5 µM, respectively.


Localized Scleroderma: A Clinical Review.
Localized scleroderma (LS) is characterized by excessive collagen deposition leading to thickening of the dermis, subcutaneous tissue or both. The outcome for most patients with localized scleroderma is directly related to the type and stage of the affected tissue. The major challenge for untreated patients is not increased mortality risk, rather deformity and growth defects from skin, muscle and bone abnormalities. Treatment is individualized to type and stage of the lesion and may include pharmacologic and non-pharmacologic therapies. Among the pharmacologic modalities, methotrexate with systemic glucocorticoids is currently the mainstay of treatment. More controlled trials are needed to determine the length of treatment and the maintenance dose of this combination therapy.


Updates in the treatment of vaginal cancer.
Vaginal cancer is a rare cancer. A lot of the data used in the treatment of this cancer are extrapolated from cervical cancer data. Radiation therapy plays a significant role in the treatment of vaginal cancer. The advances in radiation therapy in both external beam and brachytherapy have improved local control, survival, and toxicity. Brachytherapy plays an important role in treating vaginal cancer, but treatment should be individualized to each tumor. Imaging, particularly magnetic resonance imaging, plays an essential role in the management of patients with vaginal cancer, from diagnosis to staging to treatment management to surveillance.


Imbalance of Th17 and Tregs in thymoma may be a pathological mechanism of myasthenia gravis.
An imbalance in Th17 cells and Tregs may be an important cause of the pathogenesis of thymoma with myasthenia gravis (MG). In this study, 30 patients with simple thymoma and 30 patients with thymoma with MG were analyzed. Flow cytometry analysis of Th17 and Tregs in peripheral blood revealed that the percentages of Th17 in thymoma were lower than those in thymoma with MG, while the percentages of Tregs were higher than those in simple thymoma. Serum cytokine ELISA assays showed that IL-6 levels in simple thymoma were lower than those in MG patients. Further, Th17 and Tregs levels were detected by immunohistochemical double staining of thymoma tissue; the number of positive Th17 cells in thymoma with MG was higher than that in simple thymoma, while positive Tregs showed the opposite results. RORγt protein and mRNA expression in thymoma with MG were both higher than those in simple thymoma. FOXP3 protein and mRNA expression in the thymoma with MG group were lower than those in simple thymoma. The results of coculture of thymoma cells and CD4 + T cells showed that thymoma cells could promote the differentiation of Th17 cells and inhibit the Tregs. Overall, Th17 cells and related transcription factors and cytokines in thymoma with MG patients were higher than those in thymoma patients, whereas, Tregs showed the opposite results, the mechanism may be that thymoma can secrete IL6 and IL21. These findings indicated that imbalances in Th17/Tregs may account for the pathogeny of thymoma with MG.


[Determination of urinary excretion of iodine in cordarone induced hyperthyroidism].
It is well known that amiodarone, an antiarrythmic drug containing iodine, can induce in some subjects thyroid function alterations: hyperthyroidism in patients resident in iodine-deficient areas and hypothyroidism in those with sufficient iodine intake. We attempted in this study to show the possibility to select the subjects at major risk of this iodine-induced hyperthyroidism, with a simple and inexpensive method such as urinary excretion iodine assay.


Reshaping the transcriptional frontier: epigenetics and somatic cell nuclear transfer.
Somatic-cell nuclear transfer (SCNT) experiments have paved the way to the field of cellular reprogramming. The demonstrated ability to clone over 20 different species to date has proven that the technology is robust but very inefficient, and is prone to developmental anomalies. Yet, the offspring from cloned animals exhibit none of the abnormalities of their parents, suggesting the low efficiency and high developmental mortality are epigenetic in origin. The epigenetic barriers to reprogramming somatic cells into a totipotent embryo capable of developing into a viable offspring are significant and varied. Despite their intimate relationship, chromatin structure and transcription are often not uniformly reprogramed after nuclear transfer, and many cloned embryos develop gene expression profiles that are hybrids between the donor cell and an embryonic blastomere. Recent advances in cellular reprogramming suggest that alteration of donor-cell chromatin structure towards that found in an normal embryo is actually the rate-limiting step in successful development of SCNT embryos. Here we review the literature relevant to the transformation of a somatic-cell nucleus into an embryo capable of full-term development. Interestingly, while resetting somatic transcription and associated epigenetic marks are absolutely required for development of SCNT embryos, life does not demand perfection.


Layer-by-Layer Encapsulation of Probiotics for Delivery to the Microbiome.
The gastrointestinal (GI) microbiome is widely investigated for its role in many diseases. However, technologies designed for microbiome delivery are lacking. Here, a layer-by-layer (LbL) approach is reported for probiotic encapsulation to protect probiotics against GI tract insults and improve their adhesion and growth on the intestines. These advantages translate to significantly enhanced survival of LbL-probiotics in vivo.


[A case report of asynchronous bilateral non-specific segmental ureteritis].
A case of asynchronous bilateral non-specific segmental ureteritis is presented. The preoperative evaluations for this lesion were considered extremely important, because the radiological and operative findings of this disease are similar to those of ureteral cancer. This lesion has the possibility of multiplicity, especially bilateral occurrence. Conservative management of the kidney should be performed.


Frog tendon structure and its relationship with locomotor modes.
Tendon collagen fibrils are the basic force-transmitting units of the tendon. Yet, surprisingly little is known about the diversity in tendon anatomy and ultrastructure, and the possible relationships between this diversity and locomotor modes utilized. Our main objectives were to investigate: (a) the ultra-structural anatomy of the tendons in the digits of frogs; (b) the diversity of collagen fibril diameters across frogs with different locomotor modes; (c) the relationship between morphology, as expressed by the morphology of collagen fibrils and tendons, and locomotor modes. To assess the relationship between morphology and the locomotor modes of the sampled taxa we performed a principal component analysis considering body length, fibrillar cross sectional area (CSA) and tendon CSA. A MANOVA showed that differences between species with different locomotor modes were significant with collagen fibril diameter being the discriminating factor. Overall, our data related the greatest collagen fibril diameter to the most demanding locomotor modes, conversely, the smallest collagen fibril CSA and the highest tendon CSA were observed in animals showing a hopping locomotion requiring likely little absorption of landing forces given the short jump distances.


Association of Endoscopic Esophageal Variceal Ligation with Duodenal Ulcer.
To determine the frequency of duodenal ulcer (DU), as well as other clinical characteristics occurring after endoscopic variceal ligation (EVL) of the esophagus. Descriptive study. The First Affiliated Hospital of Fujian Medical University, Fuzhou, China, from April 2012 to April 2013. A total of 47 patients with esophageal varices (EVr) who had also undergone EVL and gastroscopic follow-up within 3 months of the procedure was retrospectively analyzed. The status of Helicobacter pylori(Hp) infection, Child-Pugh classification, and the grades of portal hypertensive gastropathy (PHG) were collected. Sixty EVr patients without EVL treatment, but with clinical data available, served as the control group. The frequency of DU in the EVL group (29.8%, 14/47) was higher than the control group (6.7%, 4/60) (p=0.02). Hp infection rate in EVLgroup was 19.15% (9/47), while in control group was 21.67% (13/60) (p=0.813). Hp positive rate (12.5%, 1/8) in patients exhibited new DUs after EVL was comparable to the patients without DU in the EVL group (12.1%, 4/33) (p=1.00). Patients with DU after EVL received 18.79 ±8.48 of ligating bands, while in those who did not exhibit DUs received 13.85 ±6.47 (z = -2.042, p = 0.041). Logistic regression analysis showed that the occurrence of DU was not associated with age, gender, Child-Pugh classification, or the grade of PHG (p > 0.05). Esophageal EVL is associated with a higher frequency of developing DU, which is related to a larger number of applied bands but is not correlated with Hp infection status or other variables.


[Herpetic infections and human immunodeficiency virus].
Co-infection by HIV and HSV viruses his known since AIDS epidemic has started. A better knowledge of the strong relations between the two viruses has demonstrated that HSV infection increases the rate of HIV transmission. HSV2 infection is occurring in younger people below 20 years old where incidence has much increased those last years, in teen agers with a high number of sexual partners. The increase in HIV infection and transmission in HSV positive patients leads to treat genital ulcers to decline the rate of HIV and HSV transmission.


How is the collaborative-practice competency operationalized by mental health workers?
The recovery-focused competencies currently endorsed in policy emphasize collaborative relationships between mental health workers, service users, families and communities. Based on a qualitative research methodology, multi-disciplined practitioners shared their perceptions as to how mental health workers could operationalize collaborative relationships. Two community mental health centres were the setting for three focus groups, where 16 voluntary participants contributed to focused discussions. Participants discussed the pragmatics of how they work collaboratively; identifying knowledge and attitudes that underpin their practice, and elaborating on environmental influences that impact on a collaborative approach. Findings from the study recommend the practitioner role as one of advocacy and facilitation. The collaborative approach, which is inextricably related to the quality of the practitioner-service user relationship, aids sense-making for service users of their mental health experience. This requires of practitioners the qualities of openness, expression of hope, genuineness and people-first attitude that supports the building of knowledge rather than communicating it.


Hypophysitis: A Rare Cause of Impaired Consciousness in a Middle-Aged Woman.
Hypophysitis occurs predominantly in women, usually during pregnancy or postpartum period. Hypophysitis involving the anterior pituitary often presents with adrenal insufficiency. We describe a 43-year-old woman who presented with recurrent episodes of alteration of sensorium due to hyponatremia. She had a 3-month history of amenorrhea and on evaluation had evidence of pan-hypopituitarism. The magnetic resonance images of the brain showed mild pituitary enlargement and thickened pituitary stalk with homogenous contrast enhancement indicating hypophysitis. She was treated with oral steroids and thyroxine supplements. She remained asymptomatic at 15-month follow-up on low-dose oral steroids and thyroxine supplements. Hypopituitarism due to hypophysitis resulted in recurrent episodes of hyponatremia in our patient. Our case highlights the importance of a systematic search for the etiology for hyponatremic encephalopathy.


[Periodic health examination: a collaboration between an academic institution, a private firm and general practitioners].
A Swiss private company decided to launch a healthy lifestyle-oriented medical visit for its employees. Participating general practitioners had prior training through a one-day course. A brief physical examination, cholesterol and glucose analyses were included in the consultation. Half of the employees participated in the project. Health habits were similar to the general Swiss population. Four months later, 61% reported having changed at least one lifestyle health habit in order to improve their health. Most of participants and practitioners were satisfied with this type of consultation, that confirms the interest and the feasibility of such a project.


Mineral Content and Volatile Profiling of Prunus avium L. (Sweet Cherry) By-Products from Fundão Region (Portugal).
Large amounts of Prunus avium L. by-products result from sweet cherry production and processing. This work aimed to evaluate the mineral content and volatile profiling of the cherry stems, leaves, and flowers of the Saco cultivar collected from the Fundão region (Portugal). A total of 18 minerals were determined by ICP-MS, namely 8 essential and 10 non-essential elements. Phosphorus (P) was the most abundant mineral, while lithium (Li) was detected in trace amounts. Three different preparations were used in this work to determine volatiles: hydroethanolic extracts, crude extracts, and aqueous infusions. A total of 117 volatile compounds were identified using HS-SPME/GC-MS, distributed among different chemical classes: 31 aldehydes, 14 alcohols, 16 ketones, 30 esters, 4 acids, 4 monoterpenes, 3 norisoprenoids, 4 hydrocarbons, 7 heterocyclics, 1 lactone, 1 phenol, and 2 phenylpropenes. Benzaldehyde, 4-methyl-benzaldehyde, hexanal, lilac aldehyde, and 6-methyl-5-hepten-2-one were the major volatile compounds. Differences in the types of volatiles and their respective amounts in the different extracts were found. This is the first study that describes the mineral and volatile composition of Portuguese sweet cherry by-products, demonstrating that they could have great potential as nutraceutical ingredients and natural flavoring agents to be used in the pharmaceutical, cosmetic, and food industries.


The effect of thyroid antibody positivity on reference intervals for thyroid stimulating hormone (TSH) and free thyroxine (FT4) in an aged population.
Our aims were: 1) to analyze the effect of the methodology used to derive clinically feasible cut-off values for thyroid peroxidase antibodies (TPOAb) and thyroglobulin antibodies (TgAb), which exhibit highly skewed distributions; and 2) to describe the influence of thyroid antibodies on thyroid stimulating hormone (TSH) and free thyroxine (FT4) reference intervals among thyroid disease-free aged subjects. The reference population consisted of 1086 individuals with a mean age of 73 years. The impacts of TPOAb and/or TgAb positivity on the reference intervals of TSH and FT4 were evaluated by both including and excluding subjects with elevated thyroid antibodies. The exclusion of subjects with elevated thyroid antibodies had no effect on the FT4 reference interval in either gender or on the TSH reference interval in men. Among women, the exclusion of 196 (34%) thyroid antibody-positive subjects resulted in lowering of the upper reference limit of TSH from 7.2 to 5.8 mIU/L. When the more stringent "mode-method" by summing mode+(mode-2.5th percentile) was used, 334 women (58%) were excluded and the upper reference limit of TSH remained essentially identical. Regardless of the statistical methodology used to derive cut-off values, the effect of antibody positivity was found to be less than expected.


[Two possible modes of memory trace reproduction].
The current study tested spatial memory in 8 adult domestic cats using a spatial delayed response ("DR") memory task. Two existing different variants of that task were used by us in 8 adult domestic cats. One group of 4 cats was tested in the direct variant ("DDR"), while other group of 4 cats was trained to perform indirect variant ("IDDR"). DDR was based on the visual perception of baited food wells located in different places of large room which was rich of distal landmarks; testing of DDR proceeded initially with short delay intervals (minutes) followed after attainment of correct performance criterion (80% of correct responses during two consecutive days, each encompassing 20 trials of DDR), by long delays including one hour. In parallel other four cats were tested in NDDR situation. In that case before memory testing in cats was established quite strong (100% correct) visual discrimination of two different visual stimuli of the "go left - go right" type. Afterwards actual testing of IDDR was issued just with the same program of different delay presentations as was used in preceding DDR situation. Results obtained convinced us, that results of two different methods of DR testing might be explained by different modes of trace retrieval (reproduction) from the visuospatial memory stores - one working in DDR is based on the "cue recall" aided by distal landmarks, presented in DDR situations, and the other working in NDDR based on the "cue recognition" aided by visual discrimination of visual conditioned stimuli.


Reading embodied consciousness in "Emma".
The language of Emma (1815) reflects Jane Austen's developing view of embodied consciousness and her particular interest in this novel in the physical manifestations of emotions, such as blushes and nervous responses. The discursive exploration of the inner life in Emma is the product of a cultural context that features emerging brain science and Austen's own conceptualization of the psychophysical nature of emotions. This article analyzes the language of mind and emotion in Emma, to contend that Austen grapples with the implications of the idea of embodied consciousness in a narrative that contrasts mind reading with interpreting the body.


[Relationships between the hippocampus and medial septal region and their alterations during epileptogenesis].
EEGs of the hippocampus and the medial septal region (MSR) in the control conditions and during repeated stimulation of the perforant path were simultaneously recorded in awake guinea pigs. Changes in correlation of activity of these structures during seizures provoked by the stimulation (model of acute epilepsy) and in the process of epileptogenesis induced by the kindling (model of chronic epilepsy) were analyzed. A high correlation of the baseline activities of the hippocampus and MSR observed in the control sharply decreased during acute and chronic seizures. Kindling led to emergence of the MSR capability of hippocampus-independent generation of the field seizure discharges. In the process of kindling the progressive disintegration of activities of the hippocampus and the MSR was revealed being indicative of disorders in functioning of the septohippocampal network during epileptogenesis.


GABA-immunoreactive neurons and terminals in the cat periaqueductal gray matter: a light and electron microscopic study.
Immunocytochemical and electron microscopic methods were used to study the GABAergic innervation in adult cat periaqueductal gray matter (PAG). A mouse monoclonal antibody against gamma -aminobutyric acid (GABA) was used to visualize the inhibitory neuronal system of PAG. At light microscopy, GABA-immunopositive (GABA(IP)) neurons formed two longitudinally oriented columns in the dorsolateral and ventrolateral PAG that accounted for 36% of the neuronal population of both PAG columns; their perikaryal cross-sectional area was smaller than that of unlabeled (UNL) neurons found in the same PAG subdivisions. At electron microscopic level, patches of GABA immunoreactivity were readily detected in neuronal cell bodies, proximal and distal dendrites, axons and axon terminals. Approximately 35-36% of all terminals were GABA(IP); they established symmetric synapses with dendrites (84.72% of the sample in the dorsolateral PAG and 86.09% of the sample in the ventrolateral PAG) or with cell bodies (7-10% of the sample). Moreover, 49.15% of GABA(IP) axon terminals in the dorsolateral and 52.16% in the ventrolateral PAG established symmetric synapses with GABA(IP) dendrites. Immunopositive axon terminals and unlabeled terminals were also involved in the formation of a complex synaptic arrangment, i.e. clusters of synaptic terminals in close contact between them that were often observed in the PAG neuropil. Moreover, a fair number of axo-axonic synapses between GABA(IP) and/or UNL axon terminals were present in both PAG subdivisions. Several dendro-dendritic synapses between labeled and unlabeled dendrites were also observed in both PAG subdivisions. These results suggest that in the cat PAG there exist at least two classes of GABArgic neurons. The first class could exert a tonic control on PAG projecting neurons, the second could act on those GABAergic neurons that in turn keep PAG projecting neurons under tonic inhibition. The functional implications of this type of GABAergic synapse organization are discussed in relation to the dishinibitory processes that take place in the PAG.


[Standardized diagnosis and treatment of azoospermia: a report of 1027 cases].
To constitute an appropriate procedure for clinical diagnosis and treatment for men with azoospermia in China. Following a procedure which combined the recommendations from WHO, European Association of Urology (EAU), American Urological Association (AUA), American Society for Reproductive Medicine (ASRM) with our own experiences, 1 027 patients with azoospermia were divided into different subtypes according to patients' case histories and outcomes of various routine and specialized examinations. Then appropriate treatments were chosen according to the diagnostic subtypes. There were 516 cases of obstructive azoospermia, 315 of non-obstructive azoospermia and 150 of combined (obstructive and non-obstructive) azoospermia and 46 of azoospermia with special aetiology. Of the 96 patients who adopted surgical intervention, 28 underwent transurethral resection of the ejaculatory ducts, 10 vasovasostomy and 58 vasoepididymostomy. Of the 45 patients who adopted drug intervention, 16 with inflammatory obstructive azoospermia were treated with antibiotics and 29 with hypogonadotropic hypogonadism were treated with hormonal therapy. Of all the subjects, 789 patients chose in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) and 142 chose artificial insemination of donor (AID) or adoption. (1) A standard procedure should be established for the diagnosis, classification and treatment of azoospermia; (2) Special examinations and treatments should be given to cases of azoospermia with special aetiology; (3) According to the diagnosis and the patients' conditions, the most appropriate treatment options should be recommended to them; (4) The hereditary factors in azoospermia should be seriously considered, and a genetic work-up and counseling should be offered in such cases.


Digital micromirror devices: principles and applications in imaging.
A digital micromirror device (DMD) is an array of individually switchable mirrors that can be used in many advanced optical systems as a rapid spatial light modulator. With a DMD, several implementations of confocal microscopy, hyperspectral imaging, and fluorescence lifetime imaging can be realized. The DMD can also be used as a real-time optical processor for applications such as the programmable array microscope and compressive sensing. Advantages and disadvantages of the DMD for these applications as well as methods to overcome some of the limitations will be discussed in this article. Practical considerations when designing with the DMD and sample optical layouts of a completely DMD-based imaging system and one in which acousto-optic deflectors (AODs) are used in the illumination pathway are also provided.


Active transport of biogenic amines in chromaffin granule membrane vesicles.
Chromaffin granule membrane vesicles accumulate large amounts of catecholamines against their concentration gradients. This process is ATP-dependent, reserpine, FCCP and nigericin sensitive. Carrier-mediated, reserpine-sensitive accumulation has also been demonstrated in the absence of ATP when a pH gradient (delta pH) is artificially generated across the membrane. Crude preparation of 5-hydroxytryptamine storage vesicles from rat brain or from pig platelets showed similar requirement of a transmembrane pH gradient for accumulation of the amine. The catecholamine transporter from chromaffin granules has been solubilized by the use of detergents in the presence of phospholipids. Removal of the detergent either by Sephadex filtration or by dialysis results in the formation of proteoliposomes which catalyze delta pH-dependent, reserpine-sensitive catecholamine accumulation. Under proper conditions, the solubilized H+-translocating ATPase has been incorporated into the same proteoliposomes with the catecholamine transporter, and ATP-dependent transport has been measured. The reconstituted protein shows specificity and affinity towards catecholamines similar to the native one.


Bioinformatics approach to identify the influences of SARS-COV2 infections on atherosclerosis.
Coronavirus disease (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has been a global pandemic since early 2020. Understanding the relationship between various systemic disease and COVID-19 through disease ontology (DO) analysis, an approach based on disease similarity studies, has found that COVID-19 is most strongly associated with atherosclerosis. The study provides new insights for the common pathogenesis of COVID-19 and atherosclerosis by looking for common transcriptional features. Two datasets (GSE152418 and GSE100927) were downloaded from GEO database to search for common differentially expressed genes (DEGs) and shared pathways. A total of 34 DEGs were identified. Among them, ten hub genes with high degrees of connectivity were picked out, namely C1QA, C1QB, C1QC, CD163, SIGLEC1, APOE, MS4A4A, VSIG4, CCR1 and STAB1. This study suggests the critical role played by Complement and coagulation cascades in COVID-19 and atherosclerosis. Our findings underscore the importance of C1q in the pathogenesis of COVID-19 and atherosclerosis. Activation of the complement system can lead to endothelial dysfunction. The DEGs identified in this study provide new biomarkers and potential therapeutic targets for the prevention of atherosclerosis.


Recent cell printing systems for tissue engineering.
Three-dimensional (3D) printing in tissue engineering has been studied for the bio mimicry of the structures of human tissues and organs. Now, it is being applied to 3D cell printing, which can position cells and biomaterials, such as growth factors, at desired positions in the 3D space. However, there are some challenges of 3D cell printing, such as cell damage during the printing process and the inability to produce a porous 3D shape owing to the embedding of cells in the hydrogel-based printing ink, which should be biocompatible, biodegradable, and non-toxic, etc. Therefore, researchers have been studying ways to balance or enhance the post-print cell viability and the print-ability of 3D cell printing technologies by accommodating several mechanical, electrical, and chemical based systems. In this mini-review, several common 3D cell printing methods and their modified applications are introduced for overcoming deficiencies of the cell printing process.


Selective inhibitors of vitamin D metabolism--new concepts and perspectives.
Levels of active vitamin D (VD) are controlled by synthesis via CYP27B1 and self-induced metabolism by CYP24A1. Unbalanced high CYP24A1 expression due to induction by diverse endogenous compounds and xenobiotics, and amplification found in various tumours, might lead to local VD deficiency, thereby causing/reinforcing disorders. Using primary human keratinocytes, CYP24A1 expression was examined at the mRNA level by dot-blot and Northern blot hybridization, and at the enzyme activity level by analysing HPLC profiles from incubations with 3H-labelled VD metabolites. We have developed a one-step protocol to screen test compounds for potent inhibition of CYP24A1 along with selectivity over CYP27B1 and adequate metabolic stability. These inhibitors amplified hormone levels and, thereby, its function, indicated by increased CYP24A1 expression. Moreover, they stabilized the expression of a CYP24A1 splice variant, possibly serving as a buffer of VD metabolites. In addition, a low abundant, constitutive 24-hydroxylase, active in the low nanomolar range is described. Selective CYP24A1 inhibitors could herald a new era for vitamin D research, as well as for therapeutic application. Inhibitors may be used as single entities or in combination with low doses of potent analogs to prevent and treat various defects of growth and differentiation, and neuro-immuno-endocrine disorders.


Species composition, diversity, and the abundance of arthropods inhabiting burrows of the common hamster (Cricetus cricetus L.).
The is insufficient knowledge of arthropod communities occurring in specific microhabitats. In this study, we characterize the arthropod assemblages inhabiting burrows of the common hamster (Cricetus cricetus L.) and factors that determine their diversity and abundance. We tested the following hypotheses: (1) arthropod assemblages are associated with a particular dominant vegetation occurring in the vicinity of burrows; (2) a correlation exists between fine-scale geographic distances among burrows and assemblage dissimilarity; and (3) the type of trap influences the sampling success of captured arthropods. We found 73 morphospecies belonging to 16 families in 109 burrows, most of which were in the families Staphylinidae (Coleoptera) and Parasitidae (Arachnida: Acari: Mesostigmata). The most abundant families were Staphylinidae, Cryptophagidae (Coleoptera), Parasitidae, and Macrochelidae (Mesostigmata) (78.89%). Among the identified species, we found Aleochara irmgardis (Staphylinidae) and Poecilochirus sexclavatus (Parasitidae) which had not yet been reported in Poland, and several other rare species. Meat-baited traps captured 64.34% more individuals, which were more diverse and species-rich than the non-baited control traps, but the former was more selective for saprophages, necrophages, and coprophages. The burrows located in areas overgrown by triticale (a hybrid of wheat and rye) were inhabited by 69.86% of the identified arthropod species, and these also had the highest abundance (64.07%) in comparison with other habitats. However, differences in sample size biased our results toward and overestimate arthropods associated with this vegetation. This study underlines that the species composition detected in burrows was affected by the methods used and hamster preferences for a specific habitat rather than the geographic proximity of the burrows. More extensive sampling across multiple habitats will be necessary to confirm our findings.


Ambiguous genitalia: medical, socio-cultural and religious factors affecting management in Saudi Arabia.
Twenty-eight children with ambiguous genitalia were seen at King Khalid University Hospital over a 6-year period. The incidence of this disorder was 0.4/1000 live births. Of the total, 21 (75%) were Saudis and seven (25%) were non-Saudis. The consanguinity rate was 67.9%. Twenty-four (85.7%) were born in hospital and four (14.3%) at home. In only three (10.7%) was the news first broken to the parents by a senior doctor, in 13 (46.4%) by a junior doctor, and in 11 (39.3%) by a nurse. Ambiguous genitalia were observed in 22 (78.6%) at birth and in six (21.4%) were picked up later. Owing to a lack of immediate investigative facilities and for some socio-cultural reasons, 19 of the latter groups were assigned sex without prior investigations. There was an obvious preference to assign male sex. On investigation, 13 (46.4%) had XX chromosomes, 11 (39%) XY and one (3.6%) XO: in three (10.7%), chromosomal results were not available. There were 14 cases (50%) of congenital adrenal hyperplasia, two of 5-alpha reductase deficiency (7.1%), and five of testicular feminization syndrome (17.9%), in addition to others. After investigation, five (17.9%) of the children needed sex reassignment. This was accepted by two and rejected for socio-cultural reasons by three. The opinion of the religious leaders was obtained. Some recommendations on management of these cases are made, based on our local experience.


Hypomorphic expression of parathyroid Bmal1 disrupts the internal parathyroid circadian clock and increases parathyroid cell proliferation in response to uremia.
The molecular circadian clock is an evolutionary adaptation to anticipate recurring changes in the environment and to coordinate variations in activity, metabolism and hormone secretion. Parathyroid hyperplasia in uremia is a significant clinical challenge. Here, we examined changes in the transcriptome of the murine parathyroid gland over 24 hours and found a rhythmic expression of parathyroid signature genes, such as Casr, Vdr, Fgfr1 and Gcm2. Overall, 1455 genes corresponding to 6.9% of all expressed genes had significant circadian rhythmicity. Biological pathway analysis indicated that the circadian clock system is essential for the regulation of parathyroid cell function. To study this, a novel mouse strain with parathyroid gland-specific knockdown of the core clock gene Bmal1 (PTHcre;Bmal1flox/flox) was created. Dampening of the parathyroid circadian clock rhythmicity was found in these knockdown mice, resulting in abrogated rhythmicity of regulators of parathyroid cell proliferation such as Sp1, Mafb, Gcm2 and Gata3, indicating circadian clock regulation of these genes. Furthermore, the knockdown resulted in downregulation of genes involved in mitochondrial function and synthesis of ATP. When superimposed by uremia, these PTHcre;Bmal1flox/flox mice had an increased parathyroid cell proliferative response, compared to wild type mice. Thus, our findings indicate a role of the internal parathyroid circadian clock in the development of parathyroid gland hyperplasia in uremia.


Evolutionary Dynamics of the Pericentromeric Heterochromatin in Drosophila virilis and Related Species.
Pericentromeric heterochromatin in Drosophila generally consists of repetitive DNA, forming the environment associated with gene silencing. Despite the expanding knowledge of the impact of transposable elements (TEs) on the host genome, little is known about the evolution of pericentromeric heterochromatin, its structural composition, and age. During the evolution of the Drosophilidae, hundreds of genes have become embedded within pericentromeric regions yet retained activity. We investigated a pericentromeric heterochromatin fragment found in D. virilis and related species, describing the evolution of genes in this region and the age of TE invasion. Regardless of the heterochromatic environment, the amino acid composition of the genes is under purifying selection. However, the selective pressure affects parts of genes in varying degrees, resulting in expansion of gene introns due to TEs invasion. According to the divergence of TEs, the pericentromeric heterochromatin of the species of virilis group began to form more than 20 million years ago by invasions of retroelements, miniature inverted repeat transposable elements (MITEs), and Helitrons. Importantly, invasions into the heterochromatin continue to occur by TEs that fall under the scope of piRNA silencing. Thus, the pericentromeric heterochromatin, in spite of its ability to induce silencing, has the means for being dynamic, incorporating the regions of active transcription.


Mutagenesis of AS52 cells by low concentrations of lead(II) and mercury(II)
Little is known at the molecular level concerning the genotoxic effects following the acute exposure of eukaryotic cells to low concentrations of lead (II) or mercury (II). There have been conflicting reports concerning the mutagenic potential of these heavy metals, and there have not been any studies performed to determine the molecular mechanism(s) by which these metals are mutagenic. The Chinese hamster ovary cell line, AS52, contains a stably integrated single functional copy of the Escherichia coli xanthine-guanine phosphoribosyltransferase (gpt) gene. Mutations in the gpt gene confer resistance to 6-thioguanine (TG). There was little effect on viability, as measured by relative cloning efficiency, of AS52 cells exposed to lead (II) or mercury (II) up to concentrations of 0.5 microM and 0.3 microM, respectively. However, higher concentrations of the metals caused a significant increase in cell death. There was also a dose-dependent increase in the isolation of mutants resistant to TG in treated cells when compared to non-treated controls. Concentrations of the metals as low as 0.1 microM caused a significant increase in the number of mutants resistant to TG when compared to the number of spontaneous mutants obtained in nontreated controls. While the molecular mechanism(s) by which lead and mercury (II) are genotoxic is unknown, the results of this study demonstrate that low concentrations of lead (II) and mercury (II) are mutagenic in eukaryotic cells.


Intentional rounding: its role in supporting essential care.
The King's Fund Point of Care (POC) programme aims to identify and test interventions that can improve patients' experiences of care. "Intentional rounding" is one such intervention, which is being trialled by some of the teams working with the King's Fund on the POC programme. This article explains the principles of intentional rounding and how nurses can use it to ensure patients' essential care needs are met.


Long-Term Amelioration Practices Reshape the Soil Microbiome in a Coastal Saline Soil and Alter the Richness and Vertical Distribution Differently Among Bacterial, Archaeal, and Fungal Communities.
Globally soil salinity is one of the most devastating environmental stresses affecting agricultural systems and causes huge economic losses each year. High soil salinity causes osmotic stress, nutritional imbalance and ion toxicity to plants and severely affects crop productivity in farming systems. Freezing saline water irrigation and plastic mulching techniques were successfully developed in our previous study to desalinize costal saline soil. Understanding how microbial communities respond during saline soil amelioration is crucial, given the key roles soil microbes play in ecosystem succession. In the present study, the community composition, diversity, assembly and potential ecological functions of archaea, bacteria and fungi in coastal saline soil under amelioration practices of freezing saline water irrigation, plastic mulching and the combination of freezing saline water irrigation and plastic mulching were assessed through high-throughput sequencing. These amelioration practices decreased archaeal and increased bacterial richness while leaving fungal richness little changed in the surface soil. Functional prediction revealed that the amelioration practices, especially winter irrigation with saline water and film mulched in spring, promoted a community harboring heterotrophic features. β-null deviation analysis illustrated that amelioration practices weakened the deterministic processes in structuring coastal saline soil microbial communities. These results advanced our understanding of the responses of the soil microbiome to amelioration practices and provided useful information for developing microbe-based remediation approaches in coastal saline soils.


Who will drop out and who will drop in: exercise adherence in a randomized clinical trial among patients receiving active cancer treatment.
Adherence has significantly affected the efficacy of a randomized clinical trial (RCT) to test exercise interventions. The aims of this study were to analyze exercise-related adherence patterns among patients receiving active cancer treatment and to identify factors related to exercise adherence and contamination in both the intervention and control groups. This is a secondary analysis of data from an RCT of a home-based walking intervention for patients receiving active cancer treatment. Hierarchical Poisson regression analysis was used to identify factors related to exercise adherence and exercise contamination in the exercise intervention and control groups. A total of 126 patients finished the study. Exercise adherence rate in the intervention group was 32.35%, whereas exercise contamination rate in the control group was 12.07%. Independent predictors of adherence for the exercise group were baseline physical fitness, pretreatment fatigue level, treatment-related mood disturbance, and marital status (P < .01); exercise history significantly predicted exercise contamination (P < .00) in the control group. Adherence remains an issue in an exercise RCT among patients on active cancer treatment. Adherence is related to symptom, physical function, and exercise history. Exercise researchers should consider stratifying samples based on pretreatment variables found to be significantly associated with outcome variables in this study to reduce confounding effects. Oncology clinicians can use the study findings to appropriately tailor strategies to encourage exercise adherence among patients receiving active cancer treatment so that these patients can receive the known benefits of exercise.


Subluxation of the shoulder joint in stroke patients and the influence of selected factors on the incidence of instability.
The aim of the study was to evaluate the incidence of shoulder subluxation in stroke patients and the impact of rehabilitation and presence/absence of fluid in the subhumeral and subdeltoid bursae on the occurrence of instability. The study group was composed of 182 patients after stroke, with 108 men and 74 women. 156 of the patients had suffered an ischemic stroke whereas 26 had experienced a hemorrhagic stroke; 90 patients had right-sided paresis and 92 left-sided paresis. The study variables were evaluated by performing, on two occasions, an ultrasound examination of the shoulder and an assessment of upper limb functional capacity using the Brunnström scale. 182 patients of the study group participated in an exercise programme according tothe PNF concept. There was no subluxation of the shoulder on the healthy side, while on the paretic side, subluxation occurred in 25.3% of the patients. Bursal fluid was seen more frequently on the paretic side (12.6%). A statistically significant reduction of the subluxation and bursal fluid was achieved after the rehabilitation. Subluxation occurred more frequently in patients with a known presence of fluid in the bursae and in those with poorer functional capacity of the upper limb. 1. The clinical symptoms of stroke are the cause of subluxation in the shoulder joint. 2. Subluxation occurs more frequently in patients with a known presence of fluid in the subhumeral and subdeltoid bursae and in patients with reduced functional capacity. 3. The reduction of subluxation and bursal fluid was influenced by the exercise program and improved functional capacity.


Rationale and design of the magnesium in coronaries (MAGIC) study: A clinical trial to reevaluate the efficacy of early administration of magnesium in acute myocardial infarction.
Mortality rate from ST-elevation myocardial infarction remains high in elderly patients and in patients who are not eligible for reperfusion therapy. Intravenous magnesium is a promising adjunctive treatment that may reduce reperfusion injury. Several earlier trials demonstrated a mortality rate reduction with magnesium treatment, but one large trial found no benefit. Magnesium in Coronaries (MAGIC) is a large, simple trial designed to maximize the potential treatment effect of magnesium. 10, 400 high-risk patients with ST-elevation myocardial infarction will be randomly assigned to early treatment with intravenous magnesium sulfate or matching placebo. The primary end point is death at 30 days. Information on the need for treatment of shock and sustained ventricular arrhythmias will be collected to investigate the mechanisms by which magnesium might exert a beneficial effect. If magnesium proves beneficial, its use will provide a simple, cost-effective means to improve survival in this large patient population.


Decreased E2F2 Expression Correlates with Poor Prognosis and Immune Infiltrates in Patients with Colorectal Cancer.
Growing evidence has revealed that the E2F family of transcription factor 2 (E2F2) participates in the tumorigenesis and progression of various tumors, but its role in colorectal cancer (CRC) remains largely unknown. Herein, the aim of our study was to investigate the exact role of E2F2 in CRC. The expression levels of E2F2 in CRC were appraised based on the Tumor Immune Estimate Resource (TIMER), Oncomine, The Cancer Genome Atlas (TCGA), Gene Expression Omnibus (GEO) database. The results were further confirmed using CRC tumor tissues and normal controls by experimental assays including immunohistochemistry, qRT-PCR and western blot. The survival analysis of E2F2 in CRC was analyzed using PrognoScan database and TCGA data sets. In addition, the functional roles of E2F2 were examined by Gene Set Enrichment Analysis (GSEA) and immune infiltration analysis. Our results illustrated that E2F2 was significantly downregulated in CRC samples. The low E2F2 expression in CRC was prominently correlated with N, M stage and pathological stage. Decreased E2F2 expression had an unfavorable overall survivial (OS), disease free survival (DFS), disease specific survival (DSS) and progress free interval (PFI). Multivariate cox regression showed E2F2 could be an independent prognostic factors of OS in CRC. Receiver operating characteristic (ROC) analysis showed that E2F2 may serve as a potential diagnostic biomarker for CRC patients. GSEA disclosed that E2F2 was probably involved in several pathways, including ATR pathway, ATM signalling pathway, mismatch repair, base excision repair, homologous recomibination, Fanconi Anemia pathway, multicancer invasiveness signature, and cancer stem cells. Moreover, E2F2 was significantly correlated with the infiltration level of Th2, aDC, Th17, NK CD56dim, T helper and pDC cells. The current study demonstrates that decreased E2F2 expression is closely associated with poor prognosis and immune cell infiltration in CRC, which can be a promising independent prognostic biomarker and potential treatment target for CRC.


Measurements of enzymes of collagen synthesis in rats with experimental silicosis.
The levels of two enzymes of collagen biosynthesis namely lysyl oxidase and prolyl hydroxylase were measured in the lungs of rats 3 and 6 weeks after receiving a single intratracheal instillation of silica or sterile saline. Circulating levels of lysyl oxidase were also estimated. Significant increased lung-enzyme activities were observed in the silica-exposed rats at both time intervals. Plasma levels of lysyl oxidase were also found to be raised in the silica-exposed rats. These changes were not, however, accompanied by profound pathological alterations. These results demonstrate that after exposure to silica histologically detectable fibrosis is preceded by significant changes in the activities of enzymes associated with collagen synthesis.


The evolutionary emergence of cell type-specific genes inferred from the gene expression analysis of Hydra.
Cell lineages of cnidarians including Hydra represent the fundamental cell types of metazoans and provides us a unique opportunity to study the evolutionary diversification of cell type in the animal kingdom. Hydra contains epithelial cells as well as a multipotent interstitial cell (I-cell) that gives rise to nematocytes, nerve cells, gland cells, and germ-line cells. We used cDNA microarrays to identify cell type-specific genes by comparing gene expression in normal Hydra with animals lacking the I-cell lineage, so-called epithelial Hydra. We then performed in situ hybridization to localize expression to specific cell types. Eighty-six genes were shown to be expressed in specific cell types of the I-cell lineage. An additional 29 genes were expressed in epithelial cells and were down-regulated in epithelial animals lacking I-cells. Based on the above information, we constructed a database (http://hydra.lab.nig.ac.jp/hydra/), which describes the expression patterns of cell type-specific genes in Hydra. Most genes expressed specifically in either I-cells or epithelial cells have homologues in higher metazoans. By comparison, most nematocyte-specific genes and approximately half of the gland cell- and nerve cell-specific genes are unique to the cnidarian lineage. Because nematocytes, gland cells, and nerve cells appeared along with the emergence of cnidarians, this suggests that lineage-specific genes arose in cnidarians in conjunction with the evolution of new cell types required by the cnidarians.


Association between HLA-DR4 haplotypes and tuberculin skin test response in the Aché population.
The human leukocyte antigen (HLA) system has a major role in the regulation of the immune response as it is involved in the defense against pathogens. Evidence for association with tuberculosis (TB) is more consistent for class II than for class I HLA genes. TB is important among indigenous peoples in South America, not only because of its historical role in regional depopulation, but also because it is still widespread. The aim of this study was to evaluate the association of HLA class II alleles, haplotypes and genotypes and tuberculin skin test response (TST) in 76 individuals of the Aché population. Poisson Regression was employed to assess risk genotypes. DRB1*04, DQA1*03 and DQB1*03:02 were associated with TST response in this population.


Functional cell surface expression of toll-like receptor 9 promotes cell proliferation and survival in human hepatocellular carcinomas.
Toll-like receptor 9 (TLR9) is a pattern-recognition receptor that is involved in immune signaling and plays a crucial role in cell survival through recognition of various bacterial and viral components including unmethylated CpG-DNA. TLR9 expression and function in cancer cells are not well understood. We investigated the expression of TLR9, and the function of TLR9 signaling, in hepatocellular carcinoma (HCC) cells following stimulation with CpG-oligodeoxynucleotides (ODNs). Positive immunohistochemical staining for TLR9 was observed in 85.7% of HCC tissues. Western blot analysis revealed that TLR9 was expressed both on the cell membrane and in the cytoplasm of HCC cell lines. Full-length TLR9 was predominantly expressed on the membrane rather than in the cytoplasm, whereas multiple cleaved forms of TLR9 were predominantly expressed in the cytoplasm rather than on the membrane. Cell surface stimulation of TLR9 promoted cell proliferation, and, furthermore, the TLR9 agonists, CpG-ODNs, reduced the cytotoxicity of the anti-cancer drug adriamycin (ADM) via up-regulation of apoptosis inhibitors such as survivin, Bcl-xL, XIAP and cFLIP, in HCC cell lines. Although cell surface stimulation of TLR9 did not activate either the NF-kappaB signaling pathway or the type-I IFN secretion pathway, gene chip microarray analysis indicated that TLR9 agonists closely regulated multiple oncology-related genes and transcription factors involved in tumorigenesis and cancer progression. In conclusion, our results indicate that functional cell surface expression of TLR9 in human HCC may play an important role in tumorigenesis and cancer progression.


In Silico Structure Modeling and Molecular Docking Analysis of Phosphoribosyl Pyrophosphate Amidotransferase (PPAT) with Antifolate Inhibitors.
Cancer remains one of the most serious disease worldwide. Robust metabolism is the hallmark of cancer. PPAT (phosphoribosyl pyrophosphate amidotransferase) catalyzes the first committed step of de novo purine biosynthesis. Hence PPAT, the key regulatory spot in De novo purine nucleotide biosynthesis, is an attractive and credible drug target for leukemia and other cancer therapeutics. In the present study, detailed computational analysis has been performed for PPAT protein, the key enzyme in de novo purine biosynthesis which is inhibited by many folate derivatives, hence we aimed to investigate and gauge the inhibitory effect of antifolate derivatives; lomexterol (LTX) methotrexate (LTX), and pipretixin (PTX) with human PPAT to effectively capture and inhibit De novo purine biosynthesis pathway. The sequence to structure computational approaches followed by molecular docking experiments was performed to gain insight into the inhibitory mode, binding orientation and binding affinities of selected antifolate derivatives against important structural features of PPAT. Results indicated a strong affinity of antifolate inhibitors for the conserved active site of PPAT molecule encompassing a number of hydrophobic, hydrogen bonding, Vander Waals and electrostatic interactions. Conclusively, the strong physical interaction of selected antifolate inhibitors with human PPAT suggests the selective inhibition of De novo purine biosynthesis pathway by antifolate derivatives towards cancer therapeutics.


Neonatal short bowel syndrome.
In this retrospective study the management of infants who had undergone resection of more than 50% of the small bowel as newborn infants between 1970 and 1988 was analyzed to define prognostic factors. Small bowel resections were performed for atresia (36 cases), volvulus (22 cases), gastroschisis (10 cases), necrotizing enterocolitis (11 cases), and other disorders (8 cases). Patients were classified into two groups depending on the length of residual small bowel: group 1 (n = 35) had less than 40 cm of small bowel and group 2 (n = 51) had 40 to 80 cm of residual small bowel. Patients in group 2 had significantly better survival rates than those in group 1 (92.0% vs 66.6%; p less than 0.001). The patients in group 1 who were born after 1980, when home parenteral nutrition was introduced, had better survival rates than those who were treated before 1980 (95.0% vs 65.0%; p less than 0.01). The time required for acquisition of intestinal adaptation depended on the intestinal length (average, 27.3 months for group 1 and 14 months for group 2; p less than 0.01) and on the presence or absence of the ileocecal valve. Parenteral or supportive enteral nutrition, or both, ensured normal growth in both groups. We conclude that more than 90% of infants now survive after extensive small bowel resection with parenteral nutrition and that the remaining small intestine will adapt with time. Home-based parenteral nutrition allowed children to be treated in the best psychosocial environment.


Mutations disrupting histone methylation have different effects on replication timing in S. pombe centromere.
The fission yeast pericentromere comprises repetitive sequence elements packaged into heterchromatin marked by histone H3K9 methylation and Swi6 binding. Transient disruption of Swi6 during S phase allows a period of RNA synthesis which programs the RNAi machinery to maintain histone methylation. However, Swi6 is also required for early replication timing. We show that not only Swi6 but also the chromodomain protein Chp1 are delocalized during S phase. Different from loss of swi6, mutations that disrupt histone methylation in the centromere, chp1Δ and clr4Δ, undergo early DNA replication. However, timing is modestly delayed in RNAi mutants dcr1Δ or rdp1Δ, while hrr1Δ mutants resemble swi6Δ in their replication delay. Finally, we show that recruitment of RNA polymerase II in the centromere occurs independently of replication. These different effects indicate that replication timing is not simply linked to histone methylation.


A role for the 1A and L1 rod domain segments in head domain organization and function of intermediate filaments: structural analysis of trichocyte keratin.
A dynamic model is proposed to explain how the 1A and linker L1 segments of the rod domain in intermediate filament (IF) proteins affect the head domain organization and vice versa. We have shown in oxidized trichocyte IF that the head domain sequences fold back over and interact with the rod domain. This phenomenon may occur widely in reduced IF as well. Its function may be to stabilize the 1A segments into a parallel two-stranded coiled coil or something closely similar. Under differing reversible conditions, such as altered states of IF assembly, or posttranslational modifications, such as phosphorylation etc., the head domains may no longer associate with the 1A segment. This could destabilize segment 1A and cause the two alpha-helical strands to separate. Linker L1 would thus act as a hinge and allow the heads to function over a wide lateral range. This model has been explored using the amino acid sequences of the head (N-terminal) domains of Type I and Type II trichocyte keratin intermediate filament chains. This has allowed several quasi-repeats to be identified. The secondary structure corresponding to these repeats has been predicted and a model has been produced for key elements of the Type II head domain. Extant disulfide cross-link data have been used as structural constraints. A model for the head domain structure predicts that a twisted beta-sheet region may wrap around the 1A segment and this may reversibly stabilize a coiled-coil conformation for 1A. The evidence in favor of the swinging head model for IF is discussed.


Fetal Screening for Chromosomal Abnormalities.
With more and more reproductive-aged women opting to pursue genetic screening during pregnancy, health care professionals must understand the variety of testing options available as well as the advantages and limitations of each testing option. Presently, no single screening test is universally believed to be superior because the combination of the specific test and the population being tested determines the range of potential identifiable conditions as well as the positive predictive values. As a result, pre- and posttest counseling are not always straightforward and may require discussions with multiple specialists including genetic counselors, obstetricians, and pediatricians/neonatologists. The purpose of this review is to summarize the screening options currently available to pregnant women to determine their risk of having a child affected by a chromosomal disorder. Screening for chromosomal abnormalities using ultrasonography, maternal serum analytes, cell-free DNA, and preimplantation genetic testing will be discussed here. Advances in the field, including the possible future use of cell-based noninvasive prenatal screening (NIPS) as a more accurate method for genetic screening and the incorporation of screening for copy number variants (microdeletions and duplications) into traditional cell-free NIPS will also be reviewed.


Bartholin gland carcinoma: A case report.
Bartholin gland carcinomas are rare types of tumours, accounting for less than 1% among females. This case report is of a 20-years patient in her 16th week of pregnancy who presented with pain and swelling in the left groin for two weeks. Examination showed a well-defined fixed swelling with induration in the left vulva, with an irregular margin and a diameter of about 5 cm, just at the Bartholin's gland site with two palpable ipsilateral lymph nodes. Bartholin's gland abscess was suspected. The decision was to drain the pus under general anaesthesia. During the procedure, a hard mass was felt at the left Bartholin glands region. The incision was extended, which unraveled a 5*5 cm abnormal-looking mass; it was hard and fixed to the adjacent tissues. Histopathology confirmed adenocarcinoma of Bartholin gland in stage III. The oncologist advised pregnancy termination and chemoradiation. Although surgical excision is essential for treating tumours, still, a multimodal approach can prove to be more effective.


Biopsy of a renal mass: where are we now?
To review the most recent literature concerning renal mass biopsy with special consideration to three points: variation in results related to the standard used as comparison, biopsy in small renal masses (up to 4 cm in diameter) and the case for nondiagnostic biopsy. The overall rate of failed and indeterminate biopsies shows a trend for improvement. However, selection bias and the lack of a uniform index test for comparison preclude a definitive statement. Fine-needle aspiration may equal results of core biopsy, but its role in the diagnostic algorithm is not yet defined. In-vivo accuracy decreases in small renal masses with the same limitations exposed for the overall literature on renal mass biopsy. When nondiagnostic biopsies are considered, there is a need for standardization of the nomenclature in order to compare results. Re-biopsies or surgery after a nondiagnostic biopsy shows malignancy in up to 75% of the cases of renal cell carcinoma. There is a trend in increasing interest and accuracy on the subject of percutaneous biopsy of renal masses as well as a decreasing trend in the rate of nondiagnostic biopsies. In the small renal masses, most likely to be benign, a diagnostic percutaneous biopsy may have a definitive role. However, the higher rate of nondiagnostic results in this population calls for prospective studies with standard definitions and when possible homogenous index test to properly assess the diagnostic performance of the biopsy.