Explaining the disease phenotype of intergenic SNP through predicted long range regulation.
Thousands of disease-associated SNPs (daSNPs) are located in intergenic regions (IGR), making it difficult to understand their association with disease phenotypes. Recent analysis found that non-coding daSNPs were frequently located in or approximate to regulatory elements, inspiring us to try to explain the disease phenotypes of IGR daSNPs through nearby regulatory sequences. Hence, after locating the nearest distal regulatory element (DRE) to a given IGR daSNP, we applied a computational method named INTREPID to predict the target genes regulated by the DRE, and then investigated their functional relevance to the IGR daSNP's disease phenotypes. 36.8% of all IGR daSNP-disease phenotype associations investigated were possibly explainable through the predicted target genes, which were enriched with, were functionally relevant to, or consisted of the corresponding disease genes. This proportion could be further increased to 60.5% if the LD SNPs of daSNPs were also considered. Furthermore, the predicted SNP-target gene pairs were enriched with known eQTL/mQTL SNP-gene relationships. Overall, it's likely that IGR daSNPs may contribute to disease phenotypes by interfering with the regulatory function of their nearby DREs and causing abnormal expression of disease genes.


Fluorescent Immortalized Human Adipose Derived Stromal Cells (hASCs-TS/GFP+) for Studying Cell Drug Delivery Mediated by Microvesicles.
A new tool for the drug delivery is based on the use of Mesenchymal Stromal Cells (MSCs) loaded in vitro with anti-cancer drugs. Unfortunately, the restricted lifespan of MSCs represents a significant limitation to produce them in high amounts and for long time studies. Immortalized MSCs from adipose tissue (hASCs) have been generated as good source of cells with stable features. These cells could improve the development of standardized procedures for both in vitro and preclinical studies. Furthermore they facilitate procedures for preparing large amounts of secretome containing microvesicles (MVs). We used human adipose tissue derived MSCs immortalized with hTERT+SV40 (TS) genes and transfected with GFP (hASCs-TS/GFP+). This line was investigated for its ability to uptake and release anticancer drugs. Microvesicles associated to paclitaxel (MVs/PTX) were isolated, quantified, and tested on pancreatic cancer cells. The line hASCs-TS/GFP+ maintained the main mesenchymal characters and was able to uptake and release, in active form, both paclitaxel and gemcitabine. From paclitaxel loaded hASCs-TS/GFP+ cells were isolated microvesicles in sufficient amount to inhibit "in vitro" the proliferation of pancreatic tumor cells. Our study suggests that human immortalized MSCs could be used for a large scale production of cells for mediated drug delivery. Moreover, the secretion of drug-associated MVs could represent a new way for producing new drug formulation by "biogenesis". In the context of the "advanced cell therapy procedure", the MVs/PTX production would use less resource and time and it could possibly contribute to simplification of GMP procedures.


Patient Involvement in the Health Technology Assessment Process in Taiwan.
The COVID-19 pandemic initially had a smaller impact on Taiwan than on most other industrialized countries. However, an outbreak in late April 2021 led to a sharp surge in cases from mid-May 2021. Patient involvement in the health technology assessment (HTA) process, however, was not much affected by this; virtual meetings were implemented. This descriptive paper presents an overview of patient involvement in the HTA process in Taiwan via the National Health Insurance Administration (NHIA) online submission platform, participation in appraisal committees, education programs, and cooperation with patients' organizations, and outlines its progress and challenges. The National Health Insurance Act, amended in 2013, protects patients' rights and invites them to voice their opinions, which are then presented to the relevant authority. Based on this act, various mechanisms have been developed to involve patients, caregivers, and patient organizations in both the HTA and the reimbursement process. Prior to the Pharmaceutical Benefit and Reimbursement Scheme (PBRS) Joint Committee meeting, the NHIA built an online platform that allows patients to submit their opinions, which are then incorporated into the HTA reports. The results are also discussed with patient representatives, following which the related documents are published on the NHIA website. From May 2015 to December 2020, 30 patients' insights were published before the PBRS Joint Committee meetings. Of these, 19 (63%) were related to oncology cases. In Taiwan, approaches to fostering patient engagement include the use of a platform for patients' and patients groups' input, among others. Although patient engagement is important for understanding the needs of the target patient population, challenges in ensuring timely patient engagement and provision of relevant resources remain. In addition, further efforts are needed to implement and improve the visibility of patient input in the HTA process.


Impact of HDAC Inhibitors on Protein Quality Control Systems: Consequences for Precision Medicine in Malignant Disease.
Lysine acetylation is one of the major posttranslational modifications (PTM) in human cells and thus needs to be tightly regulated by the writers of this process, the histone acetyl transferases (HAT), and the erasers, the histone deacetylases (HDAC). Acetylation plays a crucial role in cell signaling, cell cycle control and in epigenetic regulation of gene expression. Bromodomain (BRD)-containing proteins are readers of the acetylation mark, enabling them to transduce the modification signal. HDAC inhibitors (HDACi) have been proven to be efficient in hematologic malignancies with four of them being approved by the FDA. However, the mechanisms by which HDACi exert their cytotoxicity are only partly resolved. It is likely that HDACi alter the acetylation pattern of cytoplasmic proteins, contributing to their anti-cancer potential. Recently, it has been demonstrated that various protein quality control (PQC) systems are involved in recognizing the altered acetylation pattern upon HDACi treatment. In particular, molecular chaperones, the ubiquitin proteasome system (UPS) and autophagy are able to sense the structurally changed proteins, providing additional targets. Recent clinical studies of novel HDACi have proven that proteins of the UPS may serve as biomarkers for stratifying patient groups under HDACi regimes. In addition, members of the PQC systems have been shown to modify the epigenetic readout of HDACi treated cells and alter proteostasis in the nucleus, thus contributing to changing gene expression profiles. Bromodomain (BRD)-containing proteins seem to play a potent role in transducing the signaling process initiating apoptosis, and many clinical trials are under way to test BRD inhibitors. Finally, it has been demonstrated that HDACi treatment leads to protein misfolding and aggregation, which may explain the effect of panobinostat, the latest FDA approved HDACi, in combination with the proteasome inhibitor bortezomib in multiple myeloma. Therefore, proteins of these PQC systems provide valuable targets for precision medicine in cancer. In this review, we give an overview of the impact of HDACi treatment on PQC systems and their implications for malignant disease. We exemplify the development of novel HDACi and how affected proteins belonging to PQC can be used to determine molecular signatures and utilized in precision medicine.


Redistribution of Monocarboxylate 1 and 4 in Hippocampus and Spatial Memory Impairment Induced by Long-term Ketamine Administration.
The monocarboxylate transporters (MCTs) MCT1, MCT2, and MCT4 are essential components of the astrocyte-neuron lactate shuttle (ANLS), which is a fundamental element of brain energetics. Decreased expression of MCTs can induce cognitive dysfunction of the brain. In the present study, we established a mouse model of long-term ketamine administration by subjecting mice to a 6-month course of a daily intraperitoneal injection of ketamine. These mice demonstrated learning and memory deficits and a significant decline in MCT1 and MCT4 proteins in the hippocampal membrane fraction, while cytoplasmic MCT1 and MCT4 protein levels were significantly increased. In contrast, the levels of global MCT2 protein were significantly increased. Analysis of mRNA levels found no changes in MCT1/4 transcripts, although the expression of MCT2 mRNA was significantly increased. We suggest that redistribution of hippocampal MCT1 and MCT4, but not MCT2 up-regulation, may be related to learning and memory deficits induced by long-term ketamine administration.


Combined 3D-QSAR modeling and molecular docking study on multi-acting quinazoline derivatives as HER2 kinase inhibitors.
A series of new quinazoline derivatives has been recently reported as potent multi-acting histone deacetylase (HDAC), epidermal growth factor receptor (EGFR), and human epidermal growth factor receptor 2 (HER2) inhibitors. HER2 is one of the major targets for the treatment of breast cancer and other carcinomas. Three-dimensional structure-activity relationship (3D-QSAR) is a well-known technique, which is used to drug design and development. This technique is used for quantitatively predicting the interaction between a molecule and the active site of a specific target. For each 3D-QSAR study, a three-dimensional model is created from a large curve fit to find a fitting between computational descriptors and biological activity. This model could be used as a predictive tool in drug design. The best model has the highest correlation between theoretical and experimental data. Self-Organizing Molecular Field Analysis (SOMFA), a grid-based and alignment-dependent 3D-QSAR method, is employed to study the correlation between the molecular properties and HER2 inhibitory potency of the quinazoline derivatives. Before presentation of inhibitor structures to SOMFA study, conformation of inhibitors was determined by AutoDock4, HyperChem and AutoDock Vina, separately. Overall, six independent models were produced and evaluated by the statistical partial least square (PLS) analysis. Among the several generated 3D-QSARs, the best model was selected on the basis of its statistical significance and predictive potential. The model derived from the superposition of docked conformation with AutoDock Vina with reasonable cross-validated q(2) (0.767), non cross-validated r(2) (0.815) and F-test (97.22) values showed a desirable predictive capability. Analysis of SOMFA model could provide some useful information in the design of novel HER2 kinase inhibitors with better spectrum of activity.


Extraordinary biological properties of a new calcium hydroxyapatite/poly(lactide-co-glycolide)-based scaffold confirmed by in vivo investigation.
This study examined the potential of a new porous calcium hydroxyapatite scaffold covered with poly (lactide-co-glycolide) (PLGA) as a bone substitute, identifying its advantages over Geistlich Bio-Oss®, considered the gold standard, in in vivo biofunctionality investigations. Structural and morphological properties of the new scaffold were analyzed by scanning electron and atomic force microscopy. The biofunctionality assays were performed on New Zealand white rabbits using new scaffold for filling full-thickness defects of critical size. The evaluated parameters were: the presence of macrophages, giant cells, monoocytes, plasma cells, granulocytes, neoangiogenesis, fibroplasia, and the percentage of mineralization. Parallel biofunctionality assays were performed using Geistlich Bio-Oss®. The appearance of bone defects 12 weeks after the new scaffold implantation showed the presence of a small number of typical immune response cells. Furthermore, significantly reduced number of capillary buds, low intensity of fibroplasia and high degree of mineralization in a lamellar pattern indicated that the inflammation process has been almost completely overcome and that the new bone formed was in the final phase of remodeling. All biofunctionality assays proved the new scaffold's suitability as a bone substitute for applications in maxillofacial surgery. It showed numerous biological advantages over Geistlich Bio-Oss® which was reflected mainly as a lower number of giant cells surrounding implanted material and higher degree of mineralization in new formed bone.


Cecal infusion of butyrate does not alter cecal concentration of butyrate in piglets fed inulin.
Cecal or distal colonic concentration of butyrate has been used as an index of butyrate production from various fermentable carbohydrates. However, we previously found that cecal concentration of butyrate does not correlate with the rate of synthesis of butyrate in the cecal lumen. As part of a larger study of the cellular effects of cecal infusions of butyrate, we sought to rule out the null hypothesis that cecal infusion of butyrate also would not alter butyrate concentration in the cecum. Piglets (n = 10) were fed sow milk replacement formula plus inulin (3 g x L(-1)). After 6 days of oral feeding, the piglets were randomly assigned into 2 equal groups: (I) Cecal infusion of phosphate-buffered NaCl and (II) cecal infusion of butyrate (2.13 micromol x kg(-1) x min(-1)). The concentration of butyrate was measured by gas chromatography in the cecum and distal colon. There was no effect of cecal butyrate infusion on butyrate concentration (mM; I vs II) in the cecum (5.7 +/- 0.4 vs 5.3 +/- 1.1) or distal colon (3.3 +/- 0.6 vs 4.1 +/- 0.8) or on the ratio of cecal butyrate concentration to the sum of the concentrations of butyrate, acetate, propionate, and valerate (0.101 +/- 0.004 vs 0.083 +/- 0.011). There was no effect of cecal butyrate infusion on the concentration of any of these short chain fatty acids. At an entry rate into the cecum within the physiological range, butyrate had no effect on cecal or distal colonic luminal concentration of butyrate.


Genital anatomy and copulatory interactions in the broad snouted Caiman (Caiman latirostris).
The broad snouted caiman is a crocodylian native to South America that is subject to extensive conservation management in both wild and farming environments. Although reproductive behaviors like egg laying and clutch care have been examined in this species, little else is known about their copulatory system. We examined the anatomy of male and female cloacal and genital tissues ex vivo to build hypotheses of their interactions during copulation and the effects of that interaction on insemination. Male phallic glans tissues were artificially inflated to expand into their copulatory state, allowing the examination and quantification of structural changes at the gross and tissue levels. Digital reconstruction of MRI stacks yielded three-dimensional tissue compartment specific glans models of the inflated state. Silicone molds of female cloacae and oviducts in conjunction with dissection and diceCT analysis allowed us to assess internal geometry and infer how male and female features interact in copulo. We observed glans expansion within the female proctodeum would result in a copulatory lock limiting deeper intromission or retraction. Intromission and subsequent creation of the copulatory lock produces extensive clitoral compression, providing a possible mechanism for female assessment of male copulatory performance. Further, glans expansion forms a distal lumen that positions the glans tip in or near the vaginal openings. A coiled, muscular vagina provides a possible mechanism for postcopulatory sexual selection by excluding semen. Together, the complex male-female interaction supports evidence for cryptic selection by female choice, which can act as a driver of genital coevolution.


High-Fish Oil and High-Lard Diets Differently Affect Testicular Antioxidant Defense and Mitochondrial Fusion/Fission Balance in Male Wistar Rats: Potential Protective Effect of ω3 Polyunsaturated Fatty Acids Targeting Mitochondria Dynamics.
High-fat diets rich in fish oil (HFO diet, mainly ω3-PUFAs), in contrast to high-fat diets rich in lard (HL diet, mainly saturated fatty acids) have been shown to induce improvement in mitochondrial function and fusion processes associated with a reduction in reactive oxygen species production in both liver and skeletal muscle. High-fat diets may also impair testicular function, and mitochondria represent important cellular organelles with a pivotal role in reproductive function. Mitochondria are dynamic organelles that frequently undergo fission/fusion processes. A shift toward mitochondrial fusion process has been associated with improvement of mitochondrial function, as well as with ω3-PUFAs protective effects. The present study aimed to analyze the effect of chronic overfeeding (six weeks) with HFO or HL diet on testicular tissue histology, oxidative stress, antioxidant defenses, and mitochondrial fusion (mitofusin 2) and fission (dynamic related protein 1) protein. Our results showed that HFO diet induced less testicular histology impairment, oxidative stress, and apoptosis compared to a HL diet. This finding was associated with an increase in antioxidant activities and a shift toward mitochondrial fusion processes induced by HFO diet compared to HL diet, suggesting that ω3-PUFAs may act as bioactive compound targeting mitochondria dynamics to prevent testicular impairment.


IgG/IgM antiphospholipid antibodies present in the classification criteria for the antiphospholipid syndrome: a critical review of their association with thrombosis.
Essentials The clinical value of IgM antibodies in thrombotic antiphospholipid syndrome (APS) is debated. By review of literature, we reconsidered the clinical value of IgM antibodies in thrombotic APS. More significant correlations with thrombosis were found for the IgG compared to IgM isotype. Unavailability of paired IgG/IgM results hampers evaluating the added value of IgM positivity. Click to hear Dr de Groot's perspective on antiphospholipid syndrome Background Despite the update of the classification criteria for the antiphospholipid syndrome (APS), difficulties persist in the identification of patients at risk for thrombosis. Current guidelines include assays detecting IgG/IgM anti-β2 -glycoprotein I and anti-cardiolipin antibodies, although the relevance of IgM antibodies has been debated. Objectives Through a review of the literature from 2001 to 2014, we aimed to formally establish the thrombotic risk stratification potential of IgM as compared with IgG anti-phospholipid antibodies (aPLs). Patients/methods One thousand two hundred and twenty-eight articles were selected by a computer-assisted search of the literature. Of the 177 studies that met our inclusion criteria, the clinical value of IgG/IgM aPLs was established through analysis of odds ratios for thrombosis or percentage of positives in the thrombotic population. Results/conclusions We clearly found more significant correlations with thrombosis for the IgG than for the IgM isotype. Nonetheless, in a minority of studies, significant associations with thrombosis were found for IgM but not IgG antibodies. The unavailability of paired results of IgG and IgM for each separate patient hampers evaluation of the added value of isolated IgM positivity. To fully take advantage of results obtained by future studies, we strongly encourage scientists to provide all studied information per patient. We planned a large multicenter study to investigate clinical associations of isolated/combined positivity for criteria/non-criteria aPLs. Importantly, because of the presence of non-pathogenic aPLs, quantitative assays are characterized by a high false-positivity rate. Optimization of functional assays, such as thrombin generation measuring the whole scheme of coagulation, may help to reduce APS-related morbidity and mortality.


[Infusion treatment in severe dehydration (infantile toxicosis)].
Severe dehydration in infants is a common life threatening condition. The therapy of different forms of dehydration has been described. Many practical suggestions for diagnostic procedures have been added. Clinical and pathophysiological peculiarities typical of the treatment of severe infantile dehydration have been stressed.


Both the microbiome and the macrobiome can influence immune responsiveness in psoriasis.
It is debatable whether intestinal dysbiosis in autoimmune disease is a cause or a consequence of chronic inflammation, but it is known that intestinal dysbiosis in the course of the disease is accompanied by an increased number of pro-inflammatory lymphocytes in the Th17 population. Yet, little is known about the systemic implications of skin and even the intestinal microbiome for skin immunity and pathogenesis in psoriasis, which the most prevalent autoimmune disease in the Caucasian population. The pathogenesis of psoriasis is multifactorial with notable contributions from genetics and environmental factors (e.g. diet, drugs and infection). This article describes alterations in the microbiome and macrobiome, which are involved in immune regulation. The composition of the gut microbiome can dramatically affect immune development and affect susceptibility to diseases, especially autoimmune disorders such as psoriasis. Understanding the mechanisms of pathogenesis induced by the micro- and macrobiome may prove crucial for innovative future solutions in skin disease treatment.


Zilebesiran, an RNA Interference Therapeutic Agent for Hypertension.
Angiotensinogen is the sole precursor of angiotensin peptides and has a key role in the pathogenesis of hypertension. Zilebesiran, an investigational RNA interference therapeutic agent with a prolonged duration of action, inhibits hepatic angiotensinogen synthesis. In this phase 1 study, patients with hypertension were randomly assigned in a 2:1 ratio to receive either a single ascending subcutaneous dose of zilebesiran (10, 25, 50, 100, 200, 400, or 800 mg) or placebo and were followed for 24 weeks (Part A). Part B assessed the effect of the 800-mg dose of zilebesiran on blood pressure under low- or high-salt diet conditions, and Part E the effect of that dose when coadministered with irbesartan. End points included safety, pharmacokinetic and pharmacodynamic characteristics, and the change from baseline in systolic and diastolic blood pressure, as measured by 24-hour ambulatory blood-pressure monitoring. Of 107 patients enrolled, 5 had mild, transient injection-site reactions. There were no reports of hypotension, hyperkalemia, or worsening of renal function resulting in medical intervention. In Part A, patients receiving zilebesiran had decreases in serum angiotensinogen levels that were correlated with the administered dose (r = -0.56 at week 8; 95% confidence interval, -0.69 to -0.39). Single doses of zilebesiran (≥200 mg) were associated with decreases in systolic blood pressure (>10 mm Hg) and diastolic blood pressure (>5 mm Hg) by week 8; these changes were consistent throughout the diurnal cycle and were sustained at 24 weeks. Results from Parts B and E were consistent with attenuation of the effect on blood pressure by a high-salt diet and with an augmented effect through coadministration with irbesartan, respectively. Dose-dependent decreases in serum angiotensinogen levels and 24-hour ambulatory blood pressure were sustained for up to 24 weeks after a single subcutaneous dose of zilebesiran of 200 mg or more; mild injection-site reactions were observed. (Funded by Alnylam Pharmaceuticals; ClinicalTrials.gov number, NCT03934307; EudraCT number, 2019-000129-39.).


Expression of AKT1 Related with Clinicopathological Parameters in Clear Cell Renal Cell Carcinoma.
Pathways such as VEGF, EGF and mTOR are known to be one of the major mechanisms of tumorigenesis including kidney cancer. To identify potential signaling pathway proteins, we performed differential/correlation analyses of mTOR-associated genes from three public datasets. AKT1 protein, one of the PI3K/AKT/mTOR pathways, turned out to be the potential by showing a consistent discrepancy between ccRCC-associated conditions as well as strong correlation with other mTOR-associated genes across the datasets. Then, we analyzed how AKT1 alteration affects clear cell renal cell carcinoma. The pathology of 58 kidney cancer patients was constructed to analyze the relationship between the expression level of AKT1 through immunohistochemical staining and their clinicopathological data. Gender, age and TNM stage did not show significant results. AKT1 is a known oncogene. However, in this study, high expression of AKT1 showed a slight correlation with lower WHO/ISUP grade, longer recurrence-free and progression-free survival rates.


Inguinal Hernia in Athletes: Role of Dynamic Ultrasound.
Inguinal hernia is a commonly encountered cause of pain in athletes. Because of the anatomic complexity, lack of standard imaging, and the dynamic condition, there is no unified opinion explaining its underlying pathology. Athletes with persistent groin pain would have a high prevalence of inguinal hernia with dynamic ultrasound, and herniorrhaphy would successfully return athletes to activity. Case-control study. Level 3. Forty-seven amateur and professional athletes with sports-related groin pain who underwent ultrasound were selected based on history and examination. Patients with prior groin surgery or hip pathology were excluded. Clinical and surgical documentation were correlated with imaging. The study group was compared with 41 age-matched asymptomatic athletes. Ultrasound was positive for hernia with movement of bowel, bladder, or omental tissue anterior to the inferior epigastric vessels during Valsalva maneuver. The 47-patient symptomatic study group included 41 patients with direct inguinal hernias, 1 with indirect inguinal hernia, and 5 with negative ultrasound. Of 42 patients with hernia, 39 significantly improved with herniorrhaphy, 2 failed to improve after surgery and were diagnosed with adductor longus tears, and 1 improved with physical therapy. Five patients with negative ultrasound underwent magnetic resonance imaging and were diagnosed with hip labral tear or osteitis pubis. The 41-patient asymptomatic control group included 3 patients with direct inguinal hernias, 2 with indirect inguinal hernias, and 3 with femoral hernias. Inguinal hernias are a major component of groin pain in athletes. Prevalence of direct inguinal hernia in symptomatic athletes was greater than that for controls ( P < 0.001). Surgery was successful in returning these athletes to sport: 39 of 42 (93%) athletes with groin pain and inguinal hernia became asymptomatic. Persistent groin pain in the athlete may relate to inguinal hernia, which can be diagnosed with dynamic ultrasound imaging. Herniorrhaphy is successful at returning athletes to sports activity.


Traditional oral health beliefs and practices of Bulang people in Yunnan, China.
The aim of the present study was to explore traditional oral health beliefs among the Bulang ethnic minority group in China. Eighteen village leaders, chiefs, elders, and seniors in Yunnan, China were assigned to three focus groups for discussion of traditional oral health beliefs. The discussions were led by a facilitator. Transcripts were made, and data were extracted. The focus group discussions on traditional oral health beliefs addressed three themes: (a) the perception of oral health; (b) the impact of healthy teeth (oral health), in that healthy teeth essentially have an impact on physical health, emotions, and social status; and (c) toothache management, which was regarded as tooth decay (dental caries). Blackening teeth was a pain relief method. Blackened teeth were reported to be part of ethnic identity, and considered esthetically pleasing. It also indicated the marital status of women. Some Bulang people believed that healthy dentition has no spacing, pain, or functioning problems. Well-aligned dentition was associated with higher social status. Toothaches were considered to be caused by tooth worms. Consulting a monk was another way to manage a toothache. Tooth blackening was considered traditional, and effective for caries prevention.


Bacterial diversity and spoilage-related microbiota associated with freshly prepared chicken products under aerobic conditions at 4°C.
This study analyzed the bacterial diversity and spoilage-related microbiota associated with freshly prepared chicken products stored aerobically at 4°C, using "bone and chicken string," a product popular in the People's Republic of China, as the study subject. Samples collected from three different factories were tray packaged with cling film and stored at 4°C. Bacterial diversity and dominant bacteria were analyzed using PCR amplification and denaturing gradient gel electrophoresis. Combined with selective cultivation of the dominant bacteria and correlation analysis, the dominant spoilage microbiota was determined. The results showed that bacterial diversity varied with different manufacturers. Such bacteria as Acinetobacter sp., Carnobacterium sp., Rahnella sp., Pseudomonas sp., Brochothrix sp., and Weissella sp. were detected in freshly prepared chicken products during storage. And Carnobacterium sp., Pseudomonas sp., and Brochothrix sp. bacteria were the common dominant spoilage bacteria groups in most freshly prepared chicken products from different factories. Carnobacterium was, for the first time, shown to be an important contributor to the spoilage-related microflora of freshly prepared chicken products stored aerobically under refrigeration. Our work shows the bacterial diversity and dominant spoilage microbiota of freshly prepared chicken products stored aerobically under refrigeration.


Formulation and In-Vitro/Ex-Vivo Characterization of Pregelled Hybrid Alginate-Chitosan Microparticles for Ocular Delivery of Ketorolac Tromethamine.
Innovative hybrid chitosan-sodium alginate (Ch-Ag) microparticles (MPs) were fabricated using both the ionic gelation method as well as the pre-gelation technique. The hybrid Ch-Ag MPs were studied for size, zeta potential, morphology, mucoadhesion, in-vitro release, corneal permeation, and ocular irritation using lens and corneal epithelial cell lines. The average particle size ranged from 1322 nm to 396 nm. The zeta potential for the prepared formulations showed an increase with increasing Ch concentrations up to a value of >35 mV; the polydispersity index (PDI) of some optimized MPs was around 0.1. Compared to drug-free MPs, ketorolac-loaded Ch-Ag MPs demonstrated a drug proportion-dependent increase in their size. SEM, as well as TEM of KT-loaded MPs, confirmed that the formed particles were quasi-spherical to elliptical in shape. The KT release from the MPs demonstrated a prolonged release profile in comparison to the control KT solution. Further, mucoadhesion studies with porcine mucin revealed that the KT-loaded MPs had effective mucoadhesive properties, and polymeric particles were stable in the presence of mucin. Corneal permeation was studied on bovine eyes, and the results revealed that Ch-based MPs were capable of showing more sustained KT release across the cornea compared with that for the control drug solution. Conclusively, the cytotoxicity assay confirmed that the investigated MPs were non-irritant and could confer protection from direct drug irritation of KT on the ocular surface. The MTT cytotoxicity assay confirmed that KT-loaded MPs showed acceptable and reasonable tolerability with both human lens and corneal epithelial cell lines compared to the control samples.


Splenic Marginal Zone Lymphoma with Acquired von Willebrand Syndrome Diagnosed via Splenic Bleeding.
An 85-year-old woman underwent emergent splenectomy due to left abdominal pain and active bleeding in a massively enlarged spleen. The histological diagnosis was splenic marginal zone lymphoma (SMZL). A prolonged activated partial thromboplastin time (APTT) was noted, and additional tests led to the diagnosis of type 2A-like acquired von Willebrand syndrome (AVWS). An APTT cross mixing test ruled out the presence of inhibitors. She received eight courses of rituximab monotherapy. The coagulation data showed no improvement, possibly because the lymphoma showed a poor response to the treatment. AVWS rarely causes bleeding in solid organs. This is the first case of SMZL with AVWS diagnosed via splenic bleeding.


The evolutionary biology of cryptic pregnancy: A re-appraisal of the "denied pregnancy" phenomenon.
Previous research on 'denied pregnancy', i.e. lack of subjective awareness of pregnancy until the end of gestation in pregnant women, is reviewed and reinterpreted in an evolutionary biological framework. Recent epidemiological studies show that this condition has a much higher incidence than previously thought (about 1:475). Very often, bodily symptoms of pregnancy (nausea, amenorrhea and abdomen swelling) are absent or greatly reduced, and neonates tend to be underweight; in many cases, pregnancy goes undetected also by relatives and physicians. Current explanations in the clinical literature are based on psychodynamic hypotheses about pregnancy-related unconscious conflicts; the lack of symptoms is accounted for by 'somatic denial'. I argue that such psychodynamic accounts are misguided for two reasons: (1) they rest on a failure to recognize the active biological role of the fetus in determining the course of pregnancy, and (2) they ignore the many levels of mother-fetus conflict over resource allocation described by biological theories of parent-offspring conflict. Here I propose to redefine this condition as 'cryptic pregnancy', and begin to explore its possible physiological correlates and evolutionary significance. In the light of parent-offspring conflict theory, cryptic pregnancy appears to reduce the costs of pregnancy, both energetic and ecological (mobility, dependence on kin/mate, etc.), thus favoring the mother at the expense of the fetus. Reduced hCG production and/or effectiveness is likely to be involved in the process. I propose and discuss three nonexclusive evolutionary hypotheses to account for this phenomenon: (1) cryptic pregnancy could be a nonadaptive outcome of conflict resolution processes over resource allocation in pregnancy, possibly related to minor disruptions of genomic imprinting mechanisms. (2) Cryptic pregnancy could result from missed spontaneous abortions of low-quality fetuses. (3) Finally, cryptic pregnancy could be an adaptive pattern of 'forced cooperation' between mother and fetus in stressful or threatening ecological circumstances, as suggested by the reported association with elevated psychosocial stress. In case of reduced survival probability, both mother and fetus would benefit if the mother reduced investment in pregnancy in order to maximize her chances of surviving and reaching delivery.


Candida glabrata prosthesis infection following pyelonephritis and septicaemia.
Candida glabrata is a well-known cause of lower urinary tract infections. Systemic infections caused by this organism are less common, but have increased dramatically in recent years. Prosthesis infection caused by C. glabrata is extremely rare. We report a case of prosthesis failure due to C. glabrata 5 y after candidaemia and pyelonephritis caused by this organism. The same C. glabrata strain was isolated from both infections, as confirmed by the random amplified polymorphic DNA (RAPD) method.


System-wide identification of myeloid markers of TB disease and HIV-induced reactivation in the macaque model of Mtb infection and Mtb/SIV co-infection.
Mycobacterium tuberculosis (Mtb) has developed specialized mechanisms to parasitize its host cell, the macrophage. These mechanisms allow it to overcome killing by oxidative burst and persist in the wake of an inflammatory response. Mtb infection in the majority of those exposed is controlled in an asymptomatic form referred to as latent tuberculosis infection (LTBI). HIV is a well-known catalyst of reactivation of LTBI to active TB infection (ATB). Through the use of nonhuman primates (NHPs) co-infected with Mtb and Simian Immunodeficiency Virus (Mtb/SIV), we are able to simulate human progression of TB/AIDS comorbidity. The advantage of NHP models is that they recapitulate the breadth of human TB outcomes, including immune control of infection, and loss of this control due to SIV co-infection. Identifying correlates of immune control of infection is important for both vaccine and therapeutics development. Using macaques infected with Mtb or Mtb/SIV and with different clinical outcomes we attempted to identify signatures between those that progress to active infection after SIV challenge (reactivators) and those that control the infection (non-reactivators). We particularly focused on pathways relevant to myeloid origin cells such as macrophages, as these innate immunocytes have an important contribution to the initial control or the lack thereof, following Mtb infection. Using bacterial burden, C-reactive protein (CRP), and other clinical indicators of disease severity as a guide, we were able to establish gene signatures of host disease state and progression. In addition to gene signatures, clustering algorithms were used to differentiate between host disease states and identify relationships between genes. This allowed us to identify clusters of genes which exhibited differential expression profiles between the three groups of macaques: ATB, LTBI and Mtb/SIV. The gene signatures were associated with pathways relevant to apoptosis, ATP production, phagocytosis, cell migration, and Type I interferon (IFN), which are related to macrophage function. Our results suggest novel macrophage functions that may play roles in the control of Mtb infection with and without co-infection with SIV. These results particularly point towards an interplay between Type I IFN signaling and IFN-γ signaling, and the resulting impact on lung macrophages as an important determinant of progression to TB.


[The 5 factor model of childhood schizophrenia].
Forty-four first-admission patients with childhood-onset schizophrenia (age at onset < or = 14 years) were examined retrospectively for 30 clinical symptoms using the Positive and Negative Syndrome Scale (PANSS; 15). A principal component analysis with varimax rotation was applied to the full item set of this scale and revealed five orthogonal independent symptom groups: cognition affect, social withdrawal, anti-social behavior, excitement, and reality distortion. In order to validate these psychopathological dimensions we analyzed the relation between the five factor scores and outcome variables (Disability Assessment Schedule, DAS-M; 13) several years after onset: Social withdrawal was correlated with poor outcome; reality distortion was related to good outcome (P < 0.01). A multivariate ANOVA identified group differences in the anti-social behavior factor between acute and insidious onset of illness and between boys and girls; patients with an acute onset scored significantly higher on the excitement factor than those with an insidious onset (P < 0.05). According to our results more than two dimensions are necessary to describe the psychopathology of childhood-onset schizophrenia, similar to adolescent- and adult-onset schizophrenia.


Involvement of Receptor Activity-Modifying Protein 3 (RAMP3) in the Vascular Actions of Adrenomedullin in Rat Mesenteric Artery Smooth Muscle Cells.
CALCB, ADM, and ADM2 are potent vasodilators that share a seven-transmembrane GPCR, calcitonin receptor-like receptor (CALCRL), whose ligand specificity is dictated by the presence of one of the three receptor activity-modifying proteins (RAMPs). We assessed the relative pharmacologic potency of these peptides in mesenteric artery smooth muscle cells (VSMCs) and the specific RAMP that mediates the effect of ADM in VSMCs. VSMCs, with or without RAMP knockdown, were treated with CALCB, ADM, or ADM2 in the presence or absence of their antagonists, CALCB8-37, ADM22-52, and ADM217-47, respectively, to assess the relative effect of peptides on cAMP production and their pharmacologic potency. Proximity ligation assay was used to assess the specific RAMP that associates with CALCRL to mediate the actions of ADM in VSMCs. All three peptides induced cAMP generation in VSMCs and the order of their potency is CALCB > ADM > ADM2. Effects of CALCB were blocked by CALCB8-37, ADM effects were blocked by CALCB8-37 and ADM217-47 but not ADM22-52, and ADM2 effects were blocked by all three antagonists. Knockdown of RAMP2 was ineffective, whereas knockdown of RAMP3 inhibited ADM-induced cAMP production in VSMCs, suggesting involvement of RAMP3 with CALCRL to mediate ADM effects. Absence of both RAMP2 and RAMP3 further increased CALCB-induced cAMP synthesis compared to control (P < 0.05). ADM increased CALCRL and RAMP3 association and RAMP3 knockdown inhibited the interaction of ADM with CALCRL.


The serum matrix metalloproteinase-9 level is an independent predictor of recurrence after ablation of persistent atrial fibrillation.
This study investigated whether the serum matrix metalloproteinase-9 level is an independent predictor of recurrence after catheter ablation for persistent atrial fibrillation. Fifty-eight consecutive patients with persistent atrial fibrillation were enrolled and underwent catheter ablation. The serum matrix metalloproteinase-9 level was detected before ablation and its relationship with recurrent arrhythmia was analyzed at the end of the follow-up. After a mean follow-up of 12.1±7.2 months, 21 (36.2%) patients had a recurrence of their arrhythmia after catheter ablation. At baseline, the matrix metalloproteinase-9 level was higher in the patients with recurrence than in the non-recurrent group (305.77±88.90 vs 234.41±93.36 ng/ml, respectively, p=0.006). A multivariate analysis showed that the matrix metalloproteinase-9 level was an independent predictor of arrhythmia recurrence, as was a history of atrial fibrillation and the diameter of the left atrium. The serum matrix metalloproteinase-9 level is an independent predictor of recurrent arrhythmia after catheter ablation in patients with persistent atrial fibrillation.


Parkinson's disease and occupational exposure to organic solvents in Finland: a nationwide case-control study.
This study aimed to investigate the association between Parkinson's disease (PD) and occupational exposure to organic solvents generally and chlorinated hydrocarbons (CHC) in particular. We assembled a Finland-wide case-control study for birth years 1930-1950 by identifying incident PD cases from the register of Reimbursement of Medical Costs and drawing two controls per case using incidence density sampling from the Population Information System, matched on sex, birth year, and residency in Finland in 1980-2014. Occupation and socioeconomic status (SES) were identified from national censuses. We assessed cumulative occupational exposures via FINJEM job-exposure matrix. Smoking was based on occupation-specific prevalence by sex from national surveys. We estimated confounder-adjusted PD incidence rate ratios (IRR) via logistic regression and evaluated their sensitivity to errors in FINJEM through probabilistic bias analysis (PBA). Among ever-employed, we identified 17 187 cases (16.0% potentially exposed to CHC) and 35 738 matched controls. Cases were more likely to not smoke and belong to higher SES. Cumulative exposure (CE) to CHC (per 100 ppm-years, 5-year lag) was associated with adjusted IRR 1.235 (95% confidence interval 0.986-1.547), with stronger associations among women and among persons who had more census records. Sensitivity analyses did not reveal notable associations, but stronger effects were seen in the younger birth cohort (1940-1950). PBA produced notably weaker associations, yielding a median IRR 1.097 (95% simulation interval 0.920-1.291) for CHC. Our findings imply that PD is unlikely to be related to typical occupational solvent exposure in Finland, but excess risk cannot be ruled out in some highly exposed occupations.


Deep Learning Models for Stress Analysis in University Students: A Sudoku-Based Study.
Due to the phenomenon of "involution" in China, the current generation of college and university students are experiencing escalating levels of stress, both academically and within their families. Extensive research has shown a strong correlation between heightened stress levels and overall well-being decline. Therefore, monitoring students' stress levels is crucial for improving their well-being in educational institutions and at home. Previous studies have primarily focused on recognizing emotions and detecting stress using physiological signals like ECG and EEG. However, these studies often relied on video clips to induce various emotional states, which may not be suitable for university students who already face additional stress to excel academically. In this study, a series of experiments were conducted to evaluate students' stress levels by engaging them in playing Sudoku games under different distracting conditions. The collected physiological signals, including PPG, ECG, and EEG, were analyzed using enhanced models such as LRCN and self-supervised CNN to assess stress levels. The outcomes were compared with participants' self-reported stress levels after the experiments. The findings demonstrate that the enhanced models presented in this study exhibit a high level of proficiency in assessing stress levels. Notably, when subjects were presented with Sudoku-solving tasks accompanied by noisy or discordant audio, the models achieved an impressive accuracy rate of 95.13% and an F1-score of 93.72%. Additionally, when subjects engaged in Sudoku-solving activities with another individual monitoring the process, the models achieved a commendable accuracy rate of 97.76% and an F1-score of 96.67%. Finally, under comforting conditions, the models achieved an exceptional accuracy rate of 98.78% with an F1-score of 95.39%.


Dopamine induced neurodegeneration in a PINK1 model of Parkinson's disease.
Parkinson's disease is a common neurodegenerative disease characterised by progressive loss of dopaminergic neurons, leading to dopamine depletion in the striatum. Mutations in the PINK1 gene cause an autosomal recessive form of Parkinson's disease. Loss of PINK1 function causes mitochondrial dysfunction, increased reactive oxygen species production and calcium dysregulation, which increases susceptibility to neuronal death in Parkinson's disease. The basis of neuronal vulnerability to dopamine in Parkinson's disease is not well understood. We investigated the mechanism of dopamine induced cell death in transgenic PINK1 knockout mouse neurons. We show that dopamine results in mitochondrial depolarisation caused by mitochondrial permeability transition pore (mPTP) opening. Dopamine-induced mPTP opening is dependent on a complex of reactive oxygen species production and calcium signalling. Dopamine-induced mPTP opening, and dopamine-induced cell death, could be prevented by inhibition of reactive oxygen species production, by provision of respiratory chain substrates, and by alteration in calcium signalling. These data demonstrate the mechanism of dopamine toxicity in PINK1 deficient neurons, and suggest potential therapeutic strategies for neuroprotection in Parkinson's disease.


Severe pulmonary hypertension due to combined pulmonary fibrosis and emphysema: another cause of death among smokers.
In 2005, the combined pulmonary fibrosis and emphysema (CPFE) was first defined as a distinct entity, which comprised centrilobular or paraseptal emphysema in the upper pulmonary lobes, and fibrosis in the lower lobes accompanied by reduced diffused capacity of the lungs for carbon monoxide (DLCO). Recently, the fibrosis associated with the connective tissue disease was also included in the diagnosis of CPFE, although the exposure to tobacco, coal, welding, agrochemical compounds, and tire manufacturing are the most frequent causative agents. This entity characteristically presents reduced DLCO with preserved lung volumes and severe pulmonary hypertension, which is not observed in emphysema and fibrosis alone. We present the case of a 63-year-old woman with a history of heavy tobacco smoking abuse, who developed progressive dyspnea, severe pulmonary hypertension, and cor pulmonale over a 2-year period. She attended the emergency facility several times complaining of worsening dyspnea that was treated as decompensate chronic obstructive pulmonary disease (COPD). The imaging examination showed paraseptal emphysema in the upper pulmonary lobes and fibrosis in the middle and lower lobes. The echo Doppler cardiogram revealed the dilation of the right cardiac chambers and pulmonary hypertension, which was confirmed by pulmonary trunk artery pressure measurement by catheterization. During this period, she was progressively restricted to the minimal activities of daily life and dependent on caregivers. She was brought to the hospital neurologically obtunded, presenting anasarca, and respiratory failure, which led her to death. The autopsy showed signs of pulmonary hypertension and findings of fibrosis and emphysema in the histological examination of the lungs. The authors highlight the importance of the recognition of this entity in case of COPD associated with severe pulmonary hypertension of unknown cause.


The prognostic value of interim positron emission tomography scan in patients with classical Hodgkin lymphoma.
The prognostic value of interim positron emission tomography (PET) was evaluated after 2 cycles of doxorubicin, bleomycin, vinblastin and dacarbazine in classical Hodgkin lymphoma patients (n = 229), based on Deauville criteria. In early stage non-bulky disease, bulky stage II disease, advanced stage low International Prognostic Score (IPS ≤2) and advanced stage (IPS ≥3), 3-year progression-free survival rates in PET2-negative vs. PET2-positive groups were 95·9% vs. 76·9% (P < 0·0018), 83·3% vs. 20·0% (P = 0·017), 77·0% vs. 30·0% (P < 0·001) and 71·0% vs. 44·4%(P = 0·155), respectively. The outcome after positive PET2 was better than previously reported. The results from non-randomized studies of PET2-guided therapy would be valuable with careful interpretation.


[Intrahospital outbreak of enterovirus encephalitis].
The authors examined 9 children aged from 13 months to 9 years who, while being in a general hospital, contracted enteroviral encephalitis induced by Coxsackie B3 virus with a possible involvement of Coxsackie B1 virus. This report on a group hospital outbreak of enteroviral encephalitis appears to be the first of the kind in the available literature. The authors discuss the possibility of enteroviral encephalitis and Economo's encephalitis being identical in nature.


High-Performance Magnetic-core Coils for Targeted Rodent Brain Stimulations.
Objective and Impact Statement. There is a need to develop rodent coils capable of targeted brain stimulation for treating neuropsychiatric disorders and understanding brain mechanisms. We describe a novel rodent coil design to improve the focality for targeted stimulations in small rodent brains. Introduction. Transcranial magnetic stimulation (TMS) is becoming increasingly important for treating neuropsychiatric disorders and understanding brain mechanisms. Preclinical studies permit invasive manipulations and are essential for the mechanistic understanding of TMS effects and explorations of therapeutic outcomes in disease models. However, existing TMS tools lack focality for targeted stimulations. Notably, there has been limited fundamental research on developing coils capable of focal stimulation at deep brain regions on small animals like rodents. Methods. In this study, ferromagnetic cores are added to a novel angle-tuned coil design to enhance the coil performance regarding penetration depth and focality. Numerical simulations and experimental electric field measurements were conducted to optimize the coil design. Results. The proposed coil system demonstrated a significantly smaller stimulation spot size and enhanced electric field decay rate in comparison to existing coils. Adding the ferromagnetic core reduces the energy requirements up to 60% for rodent brain stimulation. The simulated results are validated with experimental measurements and demonstration of suprathreshold rodent limb excitation through targeted motor cortex activation. Conclusion. The newly developed coils are suitable tools for focal stimulations of the rodent brain due to their smaller stimulation spot size and improved electric field decay rate.


Nail Involvements as an Indicator of Skin Lesion Severity in Psoriatic Patients.
Psoriasis is a recurrent chronic inflammatory skin disease with various mild to severe clinical manifestations. The relationship between severity of the skin lesions and nail involvement has always been underestimated. Aim of the study was to evaluate the severity of skin involvement in psoriatic patients with and without nail manifestations. In this analytic cross-sectional study, patients with psoriasis referred to Razi University Hospital of Rasht from November 2015 to March 2016 were enrolled. Demographical features (i.e. age, gender) were obtained. Psoriasis severity and nail involvement criteria were assessed by Psoriasis Area and Severity Index (PASI) and Nail Psoriasis Severity Index (NAPSI), respectively. All the gathered data were analyzed by SPSS software. In this study, 71 psoriatic patients with a mean age of 39.23±17.9 years (mean ± Standard Deviation; range: 4 to 77 years old) were studied. 22 patients (31%) had nail involvements. PASI scores were 11.7±5.7 and 5.7±4.5 in the two groups with and without nail involvements, respectively (P<0.001). There were no significant differences between age, age of onset, and duration of the disease between the two groups (P>0.05).The correlation coefficient between PASI and NAPSI was 0.367, which was statistically significant (P<0.001). Based on the findings of our study, nail involvement is an important criterion in determining the severity of skin manifestations in psoriatic patients. Additionally, a high percentage of such patients probably manifest both skin and nail manifestations. Therefore it is highly recommended to consider nail involvement when evaluating psoriasis.


[Structural and functional analysis of the cortico-spinal tract in multiple sclerosis].
The study aimed at comparison of transcranial magnetic stimulation (TMS) parameters to MRI data in multiple sclerosis (MS), taking into account clinical course of the disease. TMS was conducted in 56 patients and in 10 age- and sex-matched healthy controls. According to stage of activity and variant of the course, the patients were divided into 2 groups: one at the stage of exacerbation and another--at the stage of remission with secondary progressive MS. Latency of segmental evoked motor response was similar in patients and controls. Statistically significant between-group difference or trend toward changing of some TMS parameters were found that indicates a slow-down of the impulse conducting along the cortico-spinal tract, which is characteristic of demyelinization process. The changes were more pronounced in secondary-progressive MS. According to MRI data, a square of the spinal cord on the level of the intervertebral disk center C2/C3 was significantly smaller in the group of patients with secondary progressive MS than in that with exacerbation of the disease. The TMS parameters correlated both with the level of the focal demyelinization process and with the square of the spinal cord on a level selected. It allows to estimate expression of atrophic changes caused, in its turn, not only by myelin loss but also by axon loss.


Fabrication and Characterization of 2D Layered Perovskites with a Gradient Band Gap.
Vertical gradient band-gap heterostructures of two-dimensional (2D) layered perovskites have attracted considerable research interest due to their superior optoelectronic properties and demonstrated potential for use in optical devices. However, its fabrication has been challenging. In this investigation, 2D Ruddlesden-Popper mixed halide perovskite single crystals with a vertical gradient band gap were synthesized by using a solid-state halide diffusion process. X-ray diffraction (XRD) and scanning electron microscopy (SEM) measurements after diffusion confirm that the crystalline and morphology remain intact. The transmittance and photoluminescence (PL) spectra show the formation of a vertical gradient band gap that is ascribed to gradient halide distribution through halide intermixing. The mixed halide crystal exhibits high stability with completely suppressed phase segregation in the time-dependent PL measurement. The time-resolved photoluminescence (TRPL) spectra prove that the mixed halide sample has an enhanced carrier transport due to the Förster resonance energy transfer (FRET) effect. Besides, the halide diffusion behavior is found to be different from the previously proposed "layer-by-layer" diffusion model in exfoliated crystals. The gradient band-gap structure is critical for various applications in which vertical carrier transport is demanded.


Thermodynamic analysis of allosamidin binding to a family 18 chitinase.
Inhibition of family 18 chitinases is emerging as a target for pest and fungal control as well as asthma and inflammatory therapy. One of the best known inhibitors for these enzymes is allosamidin, a natural product. While interactions of this compound with family 18 chitinases have been studied in much detail by X-ray crystallography and standard enzymology, details of the driving forces behind its tight binding remain unknown. We have studied the thermodynamics of allosamidin binding to chitinase B (ChiB), a family 18 chitinase from Serratia marcescens, using isothermal titration calorimetry. At pH 6.0, Kd is 0.16 +/- 0.04 microM, and the binding reaction is entropically driven (DeltaSr = 44 cal/K mol) with an enthalpic penalty (DeltaHr = 3.8 +/- 0.2 kcal/mol). Dissection of the entropic term shows that a favorable conformational change in the allosamidin-ChiB complex (DeltaSconf = 37 cal/K mol) is the main contributor to the reaction. At pH 8.5, Kd decreases to 0.03 muM and the binding reaction is less entropically favorable (DeltaSr = 30 cal/K mol). While the solvation entropy change (DeltaSsolv) increases from 15 cal/K mol at pH 6.0 to 46 cal/K mol at pH 8.5, DeltaSconf becomes small and negative (-8 cal/K mol) because of an enthalpy-entropy compensation. Analyses of proton transfer showed that at pH 6.0 binding of allosamidin requires deprotonation of the Asp142-Glu144 catalytic diad. At pH 8.5, the 142-144 diad is ionized in the native enzyme, relieving the deprotonation penalty of binding and explaining why binding becomes enthalpically favorable (DeltaHr = -1.2 +/- 0.2 kcal/mol).


[Construction effect of fertile cultivated layer in black soil].
The clayey farmland soil in black soil region of Northeast China, due to the existence of thicker plough pan created by unreasonable tillage, is a main limiting factor for local agricultural production. In this paper, a field experiment was conducted to study the construction effect of fertile cultivated layer on crop yield, soil physical properties, soil moisture content, and soil microbial number. After the construction of fertile cultivated layer, the soil had a thicker cultivated layer, and the crop yield was increased. Comparing with traditional tillage, applying straw and organic manure into 20-35 cm soil layer decreased soil bulk density by 9.88% and 6.20%, increased soil porosity by 9.58% and 6.02%, and enhanced soil saturated hydraulic conductivity by 167.99 and 73.78%, respectively, indicating that the construction of fertile cultivated layer could improve soil aeration and water permeability, and enhance the infiltration of rainfall. The soil moisture content and water use efficiency under the application of straw and organic manure into plough pan were higher than those under traditional tillage, and a positive correlation was observed between the moisture content in 0-35 cm soil layer and the emergence of maize seedlings. Due to the increased organic carbon source and aeration in the constructed fertile cultivated layer, soil microbial number was also increased.


Astemizole, an H1 antagonist, has no additional therapeutic effect in rheumatoid arthritis.
Mast cells are found in increased numbers in the synovium of patients with rheumatoid arthritis (RA) and may play a role in synovitis. They produce proinflammatory substances including histamine. A double blind placebo controlled study was undertaken in 60 patients with active RA using the H1 receptor blocker, astemizole, as an adjuvant to usual therapies. After 3 months of treatment there were no significant differences in clinical and laboratory variables between the groups. We conclude that H1 antagonist treatment with astemizole has no additional therapeutic effect in RA.


Social Media Use and Its Impact on Physician Review Website Ratings in Hip and Knee Arthroplasty.
The growth in social media (SM) use and consumer-driven health care has led more patients to rate surgeons on physician review websites (PRWs). This study assessed surgeon's professional SM presence and its relationship to PRW ratings. This was a cross-sectional study of the American Association of Hip and Knee Surgeons members as of June 15, 2021. The presence of SM (Facebook, Twitter, Instagram, YouTube, LinkedIn, ResearchGate, and personal professional website) and PRW (Google [G], Healthgrades [HG], and Vitals [V]) ratings were collected. Statistical analyses compared PRW ratings among surgeons who did and did not have Any SM, defined as having at least one of the following SM accounts: Facebook; Twitter; Instagram; or YouTube. Of the 2,455 surgeons, 550 (22%) had Any SM. Compared to surgeons who did not have Any SM, surgeons who had Any SM had significantly higher G, HG, and V overall scores (G:4.1 versus 3.7; HG:4.3 versus 4.1; V:4.0 versus 3.8; P < .01), number of ratings (G:36.9 versus 26.5; HG:56.8 versus 38.3; V:45.6 versus 30.9; P < .01), and number of comments (G:24.4 versus 16.4; HG:35.2 versus 22.0; V:21.5 versus 12.3; P < .01). Surgeons who had Any SM were 1.8 (1.4 to 2.3; P < .01), 1.5 (1.2 to 1.9; P < .01), and 1.5 (1.2 to 1.9; P < .01) times more likely to have a G, HG, and V score of ≥4.0, respectively, than surgeons who did not have Any SM. Surgeons who had Any SM demonstrated a significant association with higher PRW overall scores, number of ratings, and number of comments, suggesting that SM presence may increase surgeon PRW ratings.


Acute stress disorder symptoms in children and their parents after pediatric traffic injury.
The American Academy of Pediatrics highlights the important role of pediatricians in recognizing adverse child responses to tragic events, such as traffic crashes. One challenge in effectively identifying children and their parents with troubling psychological responses to trauma is that little is known about the normal range of acute psychological responses in children and their parents in the immediate aftermath of traumatic events, making identification of adverse child responses difficult. Within the first month after a traumatic event, individuals may display reexperiencing, avoidance, and hyperarousal symptoms as well as dissociation (eg, feelings of unreality or emotional numbing). The presence of these responses, collectively known as acute stress disorder (ASD), alerts providers to those who may be at risk for ongoing difficulties. For beginning to develop an evidence base to guide pediatric care providers in addressing acute traumatic responses, the aim of the current investigation was to describe systematically the range and type of symptoms of ASD in children and their parents after pediatric traffic injury. A prospective cohort study was conducted of traffic-injured children, who were 5 to 17 years of age and admitted to the hospital for treatment of injuries from traffic crashes, and their parents. All children who met eligibility criteria between July 1999 and May 2000 were invited to participate in the study. After consent/assent was obtained, children and their custodial parents (or guardians) were interviewed within 1 month after injury via a structured assessment to determine the circumstances of the crash and the presence of ASD symptoms. Relevant demographic and clinical information (eg, age, race, gender, date of injury) was abstracted from the medical records of subjects. A survey instrument to assess the presence of ASD symptoms was completed by both the child and his or her guardian. All children completed the Child Acute Stress Questionnaire, and all parents completed the Stanford Acute Stress Reaction Questionnaire. Responses were scored for the presence of dissociation, reexperiencing, avoidance, and/or hyperarousal symptoms as well as broad distress (symptoms present in every category). Symptoms of ASD were commonly observed in the children and parents. Eighty-eight percent of children and 83% of parents reported having at least 1 clinically significant symptom; this affected 90% of the families. Broad distress was observed for a large minority: 28% of children and 23% of parents. No statistically significant association was found between child broad distress and either child age (r = -0.12) or child injury severity score (r = -0.05). chi(2) analyses revealed no significant association between broad distress and child gender, child race, or mechanism of injury. No statistically significant association was found between parent broad distress and child age (r = -0.06) or child injury severity score (r = 0.09). chi(2) analyses revealed no significant association between parent broad distress and child gender or parent presence at the crash scene. Associations were found between parent broad distress and race in that fewer white parents reported broad distress. In addition, mechanism of injury was associated with parent broad distress: more parents reported broad distress when their children were involved in pedestrian-motor vehicle crashes, and fewer parents reported broad distress when their children were injured in a bicycle fall. Pediatric care providers can expect to see some ASD symptoms in most children and parents in the immediate aftermath of traffic-related injury. Brief education is appropriate to explain that these symptoms are normal reactions that are likely to resolve. If symptoms persist for >1 month or are particularly distressing in their intensity, then referral for psychological care may be necessary for treatment of posttraumatic stress disorder. Given the high prevalence of pediatric traffic crashes and the underdiagnosis of posttraumatic stress disorder, probing for recent crash exposure might be appropriate during routine child health maintenance. The following are recommendations for pediatricians: 1) routinely call the family several days and 1 to 2 weeks after a traffic injury and ask about behavioral symptoms and family function; 2) make use of the ongoing physician-patient relationship to explore symptom presence and intensity and any functional impairment in the injured child; a brief office visit with the child and parents could serve this purpose; 3) be sure to explore the effect that the child's injury has had on the family; remember that the parent's experience posttraumatic stress symptoms after pediatric traffic-related injuries and these symptoms may limit the parent's ability to support the child; 4) provide supportive care and give families the opportunity to discuss the crash and their current feelings; do not force families to talk about the crash; 5) although any child in a traffic crash or his or her parent is at risk for posttraumatic symptomatology, regardless of injury severity, particular attention should be paid to the parents of child pedestrians who are struck by motor vehicles. These parents experience posttraumatic symptoms more commonly than parents of children in other traffic crashes.


Predictors of liver failure in primary biliary cirrhosis.
The disease progression of patients with primary biliary cirrhosis (PBC) varies significantly, and the prognostic markers that identify those patients who will develop liver failure have been scarcely studied from a Chinese cohort. Aims. We aimed to determine the predictive factors of liver failure in patients with PBC. Patients who were first diagnosed as PBC with hepatic compensation between January 2007 and December 2009 were enrolled in this cohort study. Altogether 398 patients were finally included. Of these patients, 80% were women, 98% had positive antimitochondrial antibodies, and 45% had positive antinuclear antibodies (ANA). To December 2012, a total of 38 patients developed liver failure. According to the outcome, patients who developed liver failure had had higher serum concentration of baseline total bilirubin (TBil) (p = 0.013) and total bile acid (TBA) (p < 0.001), and lower concentrations of baseline total cholesterol (Tch) (p = 0.008), than patients who did not develop liver failure. Additionally, the proportion of ANA positivity was statistically different between the two groups (p = 0.009). In the established model for predicting liver failure in PBC, three variables were finally selected out, including Tch (odds ratio (OR) 0.552, 95% confidence interval (CI) 0.394-0.774, p < 0.001), TBA (OR 1.006, 95% CI 1.002-1.010, p = 0.002), and ANA (+ versus -, OR 5.518, 95% CI 1.155-26.376, p = 0.032). ANA, Tch, and TBA are predictors of liver failure in PBC.


Regulatory mechanisms of microRNAs in colorectal cancer and colorectal cancer stem cells.
Colorectal cancer (CRC) is one of the most lethal and hard-to-treat cancers in the world, which in its advanced stages, surgery and chemotherapy are the main common treatment approaches. The microRNAs (miRNAs), as novel markers for CRC detection, promote their regulatory effects via the 3'-untranslated binding region (3'-UTR) of target messenger RNA in posttranscriptional regulation of genes and also play a pivotal role in modulating resistance to chemotherapeutic agents. These small noncoding RNAs have also a critical role in CRC stem cells (CRCSCs) regulation, comprising self-renewal, differentiation, and tumorigenesis. Cancer stem cells (CSCs) are distinctive cell types inside a tumor tissue that are believed to derive from normal somatic stem cells. The CSCs have self-renewal abilities, angiogenesis, as well as specific surface markers expression characteristics. Furthermore, they are frequently criticized for tumor maintenance, treatment resistance, tumor development, and distant metastasis. In this review, we discuss the current understandings of CRCSCs and their environment with a focus on the role of miRNAs on the regulation of CSCs and their targeting application in CRC treatment.


How Does Social Support Affect the Retention Willingness of Cross-Border E-Commerce Sellers?
E-commerce research usually focuses more on how to protect consumers' rights and increase their purchase intention from the perspective of consumers. However, we still lack understanding of e-commerce sellers, especially cross-border e-commerce sellers. Based on the stimulus-body-response theory, this paper built a moderated mediation model to test the relationships among social support, perceived benefits, perceived usefulness and sellers' willingness to retain. The results show that social support has a positive impact on perceived benefits and sellers' willingness to retain; perceived benefits play a partial intermediary role between social support and sellers' willingness to retain; and perceived usefulness moderates these mediating effects. The research results further expand the perspective of e-commerce research and reveal the mechanism and boundary conditions of the influence of social support on the retention willingness of cross-border e-commerce sellers.


Genomics and public health at CDC.
Genomics is the study of the entire genome, including all genes and their interactions with each other and with the environment. The scope of public health genomics is even broader, encompassing genetic variation in populations, both human and microbial. Molecular typing of pathogens--a mainstay of infectious disease surveillance, prevention, and control--already is used to trace epidemics, provide information for vaccine development, and monitor drug resistance. Now genomic research is producing powerful new tools for public health; for example, a newly described, microchip-based method promises to diagnose influenza infection, distinguish among viruses of human or animal origin, and detect mutations that suggest increasing virulence--all in a matter of hours.


Interplay between Diffusion and Bond Cleavage Reaction for Determining Release in Polymer-Drug Conjugates.
In conjugated polymeric drug delivery systems, both the covalent bond degradation rate and the diffusion of the freely moving drug particles affect the release profile of the formulation. Using Monte Carlo simulations in spherical matrices, the release kinetics resulting from the competition between the reaction and diffusion processes is discussed. For different values of the relative bond cleavage rate, varied over four orders of magnitude, the evolution of (i) the number of bonded drug molecules, (ii) the fraction of the freely moved detached drug within the polymer matrix, and (iii) the resulting fractional release of the drug is presented. The characteristic release time scale is found to increase by several orders of magnitude as the cleavage reaction rate constant decreases. The two extreme rate-limiting cases where either the diffusion or the reaction dominates the release are clearly distinguishable. The crossover between the diffusion-controlled and reaction-controlled regimes is also examined and a simple analytical formula is presented that can describe the full dependence of the release time on the bond cleavage rate constant. This simple relation is provided simply by the sum of the characteristic time for purely diffusional release and the bond cleavage decay time, which equals the inverse of the reaction rate constant.


Quantum spin dynamics and entanglement generation with hundreds of trapped ions.
Quantum simulation of spin models can provide insight into problems that are difficult or impossible to study with classical computers. Trapped ions are an established platform for quantum simulation, but only systems with fewer than 20 ions have demonstrated quantum correlations. We studied quantum spin dynamics arising from an engineered, homogeneous Ising interaction in a two-dimensional array of (9)Be(+) ions in a Penning trap. We verified entanglement in spin-squeezed states of up to 219 ions, directly observing 4.0 ± 0.9 decibels of spectroscopic enhancement, and observed states with non-Gaussian statistics consistent with oversqueezed states. The good agreement with ab initio theory that includes interactions and decoherence lays the groundwork for simulations of the transverse-field Ising model with variable-range interactions, which are generally intractable with classical methods.


Quantitative spatial distribution of sirolimus and polymers in drug-eluting stents using confocal Raman microscopy.
Raman spectroscopy was used to differentiate each component found in the CYPHER Sirolimus-eluting Coronary Stent. The unique spectral features identified for each component were then used to develop three separate calibration curves to describe the solid phase distribution found on drug-polymer coated stents. The calibration curves were obtained by analyzing confocal Raman spectral depth profiles from a set of 16 unique formulations of drug-polymer coatings sprayed onto stents and planar substrates. The sirolimus model was linear from 0 to 100 wt % of drug. The individual polymer calibration curves for poly(ethylene-co-vinyl acetate) [PEVA] and poly(n-butyl methacrylate) [PBMA] were also linear from 0 to 100 wt %. The calibration curves were tested on three independent drug-polymer coated stents. The sirolimus calibration predicted the drug content within 1 wt % of the laboratory assay value. The polymer calibrations predicted the content within 7 wt % of the formulation solution content. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectra from five formulations confirmed a linear response to changes in sirolimus and polymer content.


Early-life family income and subjective well-being in adolescents.
Subjective well-being (SWB) in youths positively relates to family income, however its association with income during childhood is unclear. Using longitudinal data from the US Panel Study of Income Dynamics (n = 2234 adolescents, age 12-19 years), we examined whether the timing and duration of low family income in childhood was associated with adolescent SWB. We categorized family income during childhood into state-specific quintiles. Adolescent SWB was assessed using a 12-item questionnaire (score range 3-18). We used marginal structural modelling to test for sensitive periods of exposure to low income and tested cumulative effects of income by modelling the number of years spent in the poorest income quintiles. A period in early childhood (age 0-2 years) was particularly sensitive to low family income. Adolescent SWB was 1.65 (95% CI 0.40, 2.91) points lower in those who grew up in the poorest income quintiles during early childhood compared with the top quintile. Further, each childhood year spent in the poorest income quintiles was associated with a 0.10 point (95% CI 0.04, 0.16) lower SWB score in adolescence. The timing and duration of low family income in childhood both predict individual differences in adolescent SWB. Further studies are needed to clarify the mechanisms of these models and inform public policies.


Inhibition of fructose-1,6-bisphosphatase by a new class of allosteric effectors.
A highly constrained pseudo-tetrapeptide (OC252-324) further defines a new allosteric binding site located near the center of fructose-1,6-bisphosphatase. In a crystal structure, pairs of inhibitory molecules bind to opposite faces of the enzyme tetramer. Each ligand molecule is in contact with three of four subunits of the tetramer, hydrogen bonding with the side chain of Asp187 and the backbone carbonyl of residue 71, and electrostatically interacting with the backbone carbonyl of residue 51. The ligated complex adopts a quaternary structure between the canonical R- and T-states of fructose-1,6-bisphosphatase, and yet a dynamic loop essential for catalysis (residues 52-72) is in a conformation identical to that of the T-state enzyme. Inhibition by the pseudo-tetrapeptide is cooperative (Hill coefficient of 2), synergistic with both AMP and fructose 2,6-bisphosphate, noncompetitive with respect to Mg2+, and uncompetitive with respect to fructose 1,6-bisphosphate. The ligand dramatically lowers the concentration at which substrate inhibition dominates the kinetics of fructose-1,6-bisphosphatase. Elevated substrate concentrations employed in kinetic screens may have facilitated the discovery of this uncompetitive inhibitor. Moreover, the inhibitor could mimic an unknown natural effector of fructose-1,6-bisphosphatase, as it interacts strongly with a conserved residue of undetermined functional significance.


Microprobe XRF mapping and XAS investigations of the intracellular metabolism of arsenic for understanding arsenic-induced toxicity.
Arsenic (As) is responsible for mass-poisonings worldwide following the ingestion of As-contaminated drinking water. Importantly, however, As toxicity is exploited in the antileukemia drug, Trisenox (As2O3), which successfully cures 65-80% of patients suffering relapsed acute promyelocytic leukemia. In an effort to determine the intracellular organelle and biomolecular targets of As, microprobe X-ray fluorescence (XRF) and X-ray absorption spectroscopy (XAS) analyses were performed on HepG2 cells following their exposure to high doses of arsenite (1 mM) or arsenate (20 mM). Microprobe XRF elemental mapping of thin-sectioned cells showed As accumulation in the euchromatin region of the cell nucleus (following arsenite exposure) synonymous with As targeting of DNA or proteins involved in DNA transcription. X-ray absorption near edge spectroscopy (XANES) and extended X-ray absorption fine structure (EXAFS) analysis of arsenite-treated cells, however, showed the predominance of an As tris-sulfur species, providing increased credence to As interactions with nuclear proteins as a key factor in As-induced toxicity.


[Intermediate results of aortic valve replacement using 19 mm bioprosthetic valve].
Pre- and postoperative hemodynamic parameters and activity of daily life were reviewed to estimate the effectiveness of the valve in 12 cases of single aortic valve replacement (AVR) using 19 mm bioprosthesis. All implanted prostheses were stented-valves. Carpentier-Edwards pericardial valve was used in 7 cases and Mosaic valve in 5. Left ventricular mass index (LVMI) significantly decreased from 167 +/- 36 to 133 +/- 27 g/m2 in the early postoperative period, and to 115 +/- 24 g/m2 in the intermediate phase. However, postoperative LVMI remained higher in patients with body surface area (BSA) over 1.5 m2 than in those under 1.5 m2. Postoperative activity indicated by New York Heart Association (NYHA) grade significantly improved from 2.3 +/- 1.1 to 1.4 +/- 0.5. These results indicates usefulness of 19 mm bioprosthetic valve for reducing left ventricular hypertrophy and improving activity, especially in patients with BSA smaller than 1.5 m2.


Lithium chloride promotes host resistance against Pseudomonas aeruginosa keratitis.
To explore the role of lithium chloride (LiCl) in Pseudomonas aeruginosa (PA) keratitis. B6 mice were subconjunctivally injected with LiCl in contrast to appropriate control sodium chloride (NaCl), and then routinely infected with PA. Clinical score, slit-lamp photography, hematoxylin and eosin (H&E) staining, and bacterial plate counts were used to determine the role of LiCl in PA keratitis. Messenger ribonucleic acid and protein levels of inflammatory cytokines in PA-challenged mouse corneas and in vitro cultured macrophages and neutrophils were measured with real-time PCR and enzyme-linked immunosorbent assay (ELISA), respectively. Apoptosis of the infiltrating inflammatory cells in the PA-infected murine corneas was assessed using terminal deoxynucleotidyl transferase-mediated uridine 5'-triphosphate-biotin nick end labeling staining and propidium iodide staining associated with flow cytometry. In cultured murine macrophages and neutrophils, cell apoptosis was determined with annexin V/propidium iodide double staining associated with flow cytometry and western blot analysis for cleaved caspase-3 and cleaved poly(ADP-ribose) polymerase. Treatment with LiCl reduced the severity of corneal disease by reducing corneal inflammatory response and bacterial burden. Moreover, LiCl increased anti-inflammatory cytokine interleukin-10 levels, decreased proinflammatory cytokine tumor necrosis factor-α levels, and enhanced apoptosis of infiltrating macrophages and neutrophils in the PA-infected mouse corneas. In vitro studies further confirmed that LiCl elevated anti-inflammatory cytokine expression but reduced proinflammatory cytokine production, as well as promoted cell apoptosis in murine macrophages and neutrophils. This study demonstrates a protective role of LiCl in PA keratitis. LiCl promotes host resistance against PA infection by suppressing inflammatory responses, enhancing inflammatory cell apoptosis, and promoting bacterial clearance.


Validation of automated data abstraction for SCCM discovery VIRUS COVID-19 registry: practical EHR export pathways (VIRUS-PEEP).
The gold standard for gathering data from electronic health records (EHR) has been manual data extraction; however, this requires vast resources and personnel. Automation of this process reduces resource burdens and expands research opportunities. This study aimed to determine the feasibility and reliability of automated data extraction in a large registry of adult COVID-19 patients. This observational study included data from sites participating in the SCCM Discovery VIRUS COVID-19 registry. Important demographic, comorbidity, and outcome variables were chosen for manual and automated extraction for the feasibility dataset. We quantified the degree of agreement with Cohen's kappa statistics for categorical variables. The sensitivity and specificity were also assessed. Correlations for continuous variables were assessed with Pearson's correlation coefficient and Bland-Altman plots. The strength of agreement was defined as almost perfect (0.81-1.00), substantial (0.61-0.80), and moderate (0.41-0.60) based on kappa statistics. Pearson correlations were classified as trivial (0.00-0.30), low (0.30-0.50), moderate (0.50-0.70), high (0.70-0.90), and extremely high (0.90-1.00). The cohort included 652 patients from 11 sites. The agreement between manual and automated extraction for categorical variables was almost perfect in 13 (72.2%) variables (Race, Ethnicity, Sex, Coronary Artery Disease, Hypertension, Congestive Heart Failure, Asthma, Diabetes Mellitus, ICU admission rate, IMV rate, HFNC rate, ICU and Hospital Discharge Status), and substantial in five (27.8%) (COPD, CKD, Dyslipidemia/Hyperlipidemia, NIMV, and ECMO rate). The correlations were extremely high in three (42.9%) variables (age, weight, and hospital LOS) and high in four (57.1%) of the continuous variables (Height, Days to ICU admission, ICU LOS, and IMV days). The average sensitivity and specificity for the categorical data were 90.7 and 96.9%. Our study confirms the feasibility and validity of an automated process to gather data from the EHR.


Ellipticity dependence of the third-order nonlinear optical response of graphene irradiated by two-color lights.
We theoretically investigate the third-order optical response of graphene irradiated by two-color lights, paying special attention to the quantum interference. Our studies reveal that the interplay between harmonic generation and four-wave mixing leads to unusual nonlinear optical properties with nontrivial ellipticity dependence. It is shown that, unlike the case of monochromatic light with frequency ω 0 (where the intensity of the third order response (I(3ω 0)) decreases monotonically with the ellipticity and becomes zero for circularly polarized light), I(3ω 0) may increase monotonically with the ellipticity, or be nearly independent on the ellipticity. In particular, under suitable condition, there exists an optimal nonzero ellipticity for the highest I(3ω 0). Moreover, circularly polarized third-order optical response can be generated. We have found effective methods of modulating the nonlinear optical processes, which may have various applications in optical devices based on two dimensional materials.


The interface between child/adolescent and adult mental health services: results from a European 28-country survey.
Transition-related discontinuity of care is a major socioeconomic and societal challenge for the EU. The current service configuration, with distinct Child and Adolescent Mental Health (CAMHS) and Adult Mental Health Services (AMHS), is considered a weak link where the care pathway needs to be most robust. Our aim was to delineate transitional policies and care across Europe and to highlight current gaps in care provision at the service interface. An online mapping survey was conducted across all 28 European Countries using a bespoke instrument: The Standardized Assessment Tool for Mental Health Transition (SATMEHT). The survey was directed at expert(s) in each of the 28 EU countries. The response rate was 100%. Country experts commonly (12/28) reported that between 25 and 49% of CAMHS service users will need transitioning to AMHS. Estimates of the percentage of AMHS users aged under 30 years who had has previous contact with CAMHS were most commonly in the region 20-30% (33% on average).Written policies for managing the interface were available in only four countries and half (14/28) indicated that no transition support services were available. This is the first survey of CAMHS transitional policies and care carried out at a European level. Policymaking on transitional care clearly needs special attention and further elaboration. The Milestone Study on transition should provide much needed data on transition processes and outcomes that could form the basis for improving policy and practice in transitional care.


The use of layer by layer self-assembled coatings of hyaluronic acid and cationized gelatin to improve the biocompatibility of poly(ethylene terephthalate) artificial ligaments for reconstruction of the anterior cruciate ligament.
In this study layer by layer (LBL) self-assembled coatings of hyaluronic acid (HA) and cationized gelatin (CG) were used to modify polyethylene terephthalate (PET) artificial ligament grafts. Changes in the surface properties were characterized by scanning electron microscopy, attenuated total reflection Fourier transform infrared spectroscopy, energy-dispersive X-ray spectroscopy, and contact angle and biomechanical measurements. The cell compatibility of this HA-CG coating was investigated in vitro on PET films seeded with human foreskin dermal fibroblasts over 7days. The results of our in vitro studies demonstrated that the HA-CG coating significantly enhanced cell adhesion, facilitated cell growth, and suppressed the expression of inflammation-related genes relative to a pure PET graft. Furthermore, rabbit and porcine anterior cruciate ligament reconstruction models were used to evaluate the effect of this LBL coating in vivo. The animal experiment results proved that this LBL coating significantly inhibited inflammatory cell infiltration and promoted new ligament tissue regeneration among the graft fibers. In addition, the formation of type I collagen in the HA-CG coating group was much higher than in the control group. Based on these results we conclude that PET grafts coated with HA-CG have considerable potential as substitutes for ligament reconstruction.


External auditory canal stenosis.
Stenosis of the external auditory canal may occur from a variety of causes. The numerous surgical techniques illustrate that repair may be difficult. The problems were resolved in two cases with the use of a silicone-rubber insert that is made at the time of the surgical procedure and is used throughout the postoperative period.


Adsorbed para-nitrophenol on HDTMAB organoclay--a TEM and infrared spectroscopic study.
para-Nitrophenol adsorbed on hexadecyltrimethylammonium bromide modified montmorillonite has been studied using a combination of X-ray diffraction TEM and infrared spectroscopy. Upon formation of the organoclay, the properties change from hydrophilic to hydrophobic. It is proposed that para-nitrophenol is adsorbed onto the water in the cation hydration sphere of the organoclay. As the cation is replaced by the surfactant molecules the para-nitrophenol replaces the surfactant molecules in the clay interlayer. Significant changes in the water vibrations occur in this process. Bands attributed to CH stretching and bending vibrations in general decrease as the concentration of the surfactant (CEC) increases up to 1.0 CEC. After this concentration the bands increase approaching a value the same as that of the surfactant. Strong changes occur in the HCH deformation modes of the methyl groups of the surfactant. These changes are attributed to the methyl groups locking into the siloxane surface of the montmorillonite. Such a concept is supported by changes in the SiO stretching bands of the montmorillonite siloxane surface. This study demonstrates that para-nitrophenol will penetrate into the untreated clay interlayer and replace the intercalated surfactant in surfactant modified clay, resulting in the change of the arrangement of the intercalated surfactant.


Multiple organ gigantism caused by mutation in VmPPD gene in blackgram (Vigna mungo).
Seed size is one of the most important traits in leguminous crops. We obtained a recessive mutant of blackgram that had greatly enlarged leaves, stems and seeds. The mutant produced 100% bigger leaves, 50% more biomass and 70% larger seeds though it produced 40% less number of seeds. We designated the mutant as multiple-organ-gigantism (mog) and found the mog phenotype was due to increase in cell numbers but not in cell size. We also found the mog mutant showed a rippled leaf (rl) phenotype, which was probably caused by a pleiotropic effect of the mutation. We performed a map-based cloning and successfully identified an 8 bp deletion in the coding sequence of VmPPD gene, an orthologue of Arabidopsis PEAPOD (PPD) that regulates arrest of cell divisions in meristematic cells. We found no other mutations in the neighboring genes between the mutant and the wild type. We also knocked down GmPPD genes and reproduced both the mog and rl phenotypes in soybean. Controlling PPD genes to produce the mog phenotype is highly valuable for breeding since larger seed size could directly increase the commercial values of grain legumes.


Effect of doxorubicin on mouse hybridoma B cells: stimulation of immunoglobulin synthesis and secretion.
The purpose of these studies was to investigate whether the cell-differentiating effect of anthracyclines can trigger an over-production and secretion of molecules that may interfere with the tumor-host relationship. We exposed mouse hybridoma B-cells, which are devoted to immunoglobulin production, to doxorubicin (10-40 ng/ml). We found that most doxorubicin-treated cells secreted 3- to 5-fold higher amounts of immunoglobulin than untreated cells, along with an accumulation of 50% of them in the G2+M phase of the cell cycle. The antigenic specificity of the immunoglobulin and its size pattern as determined by polyacrylamide gel electrophoresis were similar whether or not cells were treated with doxorubicin. The enhancement of immunoglobulin secretion by doxorubicin was associated with an increase of the intracellular pool of heavy and light chains of the immunoglobulin. Furthermore, an elevated synthesis of immunoglobulin was observed. The synthesis of other proteins also appeared to be modified in these circumstances. These data suggest that doxorubicin can potentiate the biological functions of target cells when used at low concentrations, elevating the production and secretion of effector molecules that interfere with the tumor-host relationship.


ConFind: a robust tool for conserved sequence identification.
ConFind (conserved region finder) identifies regions of conservation in multiple sequence alignments that can serve as diagnostic targets. Designed to work with a large number of closely related, highly variable sequences, ConFind provides robust handling of alignments containing partial sequences and ambiguous characters. Conserved regions are defined in terms of minimum region length, maximum informational entropy (variability) per position, number of exceptions allowed to the maximum entropy criterion and the minimum number of sequences that must contain a non-ambiguous character at a position to be considered for inclusion in a conserved region. Comparison of the calculated entropy for an alignment of 95 influenza A hemagglutinin sequences with random deletions results in a 98% reduction in the average error in ConFind relative to the 'Find Conserved Regions' option in BioEdit. ConFind requires Python 2.3, but Python 2.4 or an upgrade of the optparse module to Optik 1.5 is suggested. The program is known to run under Linux and DOS.


Sensorimotor and Physiological Indicators of Impairment in Mild Traumatic Brain Injury: A Meta-Analysis.
To systematically review the literature with meta-analysis to determine whether persistence of sensorimotor or physiological impairment exists between 4 weeks to 6 months post mild traumatic brain injury (mTBI), and assign level of evidence to findings. The databases PubMed, pscyINFO, SPORTdiscus, Medline, CINAHL and Embase were searched from inception to November 2016 using a priori inclusion criteria. Critical appraisal was performed, and an evidence matrix established level of evidence. Meta-analysis of pooled results identified standardized mean difference (SMD) and 95% confidence intervals (95% CI) between mTBI and healthy controls for a variety of physiological and sensorimotor indicators. Eighteen eligible articles, with a mean quality score of 15.67 (SD = 2.33) were included in the final review. Meta-analysis of center of motion variable; maximal mediolateral center of motion/center of pressure separation distance SMD [95% CI] approached significance at (-0.42 [-0.84, -0.00], I2 = 0%) for dual task, level walking indicating a potential reduction in maximal mediolateral excursion during gait in the mTBI group compared to healthy controls. Significantly reduced variability in the standard deviation of heart beat intervals was observed in the mTBI group (-0.51 [-0.74, -0.28], I2 = 0%). Overall, significant group differences in 36 sensorimotor and physiological variables (eg, balance, gait velocity and motion analysis outcomes, various oculomotor tasks, as well as heart rate variability frequency domains) were identified. Findings demonstrate that persistence of sensorimotor and physiological changes beyond expected recovery times following subacute mTBI in an adult population is possible. These findings have implications for post-injury assessment and management.