Utility of parametrectomy for early stage cervical cancer treated with radical hysterectomy.
Removal of the parametrial soft tissue is recommended for patients with cervical cancer undergoing radical hysterectomy. Parametrectomy results in significant morbidity. The objective of the study was to determine factors predictive of parametrial tumor spread and to define a subset of patients at low risk for parametrial disease. Patients with invasive cervical cancer who underwent radical hysterectomy from 1989-2005 were examined. Analysis was performed to determine factors associated with parametrial tumor spread. Survival estimates were determined using the Kaplan-Meier method. A total of 594 patients were identified. Parametrial metastases were documented in 64 (10.8%). Factors associated with parametrial disease were: histology, advanced grade, deep cervical invasion, lymphovascular space invasion (LVSI), large tumor size, advanced stage, uterine or vaginal involvement, and pelvic or para-aortic lymph node metastases (P < .0001 for each). Parametrial metastases were associated with increased risk of recurrence and decreased disease-free and overall survivals (P < .0001). A subgroup analysis was performed to identify patients at low risk for parametrial spread. In pelvic node-negative women parametrial disease was noted in 6.0% (30/498) compared with 47.9% (34 of 71) of those with positive pelvic nodes (P < .0001). If further stratified to women with negative nodes, no LVSI, and tumors < 2 cm, the incidence of parametrial disease was only 0.4%. Parametrial spread is a strong predictor of recurrence and decreased survival. Parametrial invasion is rare in patients with small tumors, no LVSI, and negative pelvic nodes (no poor prognostic factors). Further study is warranted to determine the feasibility of omitting parametrectomy in these low-risk patients.


[Intramural esophageal dissection].
The intramural esophageal dissection is a rare condition with benign course, good prognosis and low mortality. It is difficult to differentiate it from an esophageal perforation at the presentation time. The conservative therapy should be considered as the treatment of choice in most cases. Complications such as esophageal perforation, massive bleeding or persistence of the symptoms are well accepted indications for surgical procedure. Radical treatment in the form of esophagectomy is extremely rare and it is usually indicated only for patients with complicated esophageal perforation. The authors offer two cases with female patients suffering from the intramural esophageal dissection who were indicated for esophagectomy without any proven esophageal perforation.


[Reconstruction of thumb length by "on-top plasty"].
We report the case of a 23 year old man who suffered amputation of thumb and index finger at the metacarpophalangeal joint by an injury with electrical current. In order to elongate the first ray an on-top plasty of the second on the first metacarpal bone was performed. The second metacarpal was severed above the base and with the intact vessels and the first dorsal interosseous muscle placed on the distal end of the first metacarpal. The second dorsal interosseous muscle was severed. The first and second metacarpal bones united with an intramedullary cortical inlay graft. Nerve and vessels as well as the skin of the back of the hand could be moved without tension. An abdominal tube was applied to the palm because of severe scarring. As a final result pinch grip between the thumb ray and the rest fingers is possible. The other fingers are freely movable.


Mechanisms of NLRP3 inflammasome in rheumatoid arthritis and osteoarthritis and the effects of traditional Chinese medicine.
It has been widely reported that various anti-rheumatic traditional Chinese medicines (TCMs) ameliorate rheumatoid arthritis (RA) and osteoarthritis (OA) through regulating the abnormal production, assembly, and activation of the NOD-like receptor thermal protein domain-associated protein 3 (NLRP3) inflammasome. These TCMs include monomers isolated from Chinese herbs, extracts of Chinese herbs, and Chinese medical formulae with a lengthy application history. This review aimed to summarize and analyze the published articles about the NLRP3 inflammasome and its role in the pathogenesis of RA and OA. We also reviewed existing knowledge on the therapeutic mechanism of TCMs in RA and OA via the regulation of the NLRP3 inflammasome. We searched for relevant articles with the keywords "NLRP3 inflammasome", "traditional Chinese medicine," "Chinese herbal drugs," "rheumatoid arthritis," and "osteoarthritis." The information retrieval was conducted in medical Chinese and English databases from the date of construction to April 19, 2023, including PubMed, MEDLINE, Web of Science, Scopus, Ovid, China National Knowledge Infrastructure (CNKI), Chinese Biomedicine Literature Database (CBM), Chinese Science and Technology Periodicals Database (VIP), and China Online Journals (COJ). According to retrieval results, 35 TCMs have been demonstrated to relieve RA by targeting the NLRP3 inflammasome, including six traditional Chinese prescriptions, seven extracts of Chinese herbs, and 22 monomers extracted from traditional Chinese herbs and formulae. Additionally, 23 TCMs have shown anti-OA effects with abilities to modulate the NLRP3 inflammasome, including five traditional Chinese prescriptions, one extract of Chinese herbs, and 17 monomers from Chinese herbs. We summarized mechanism research about the pivotal roles of the NLRP3 inflammasome in the pathogenesis of RA and OA. Moreover, a review of TCMs with targets of the NLRP3 inflammasome in RA and OA treatment was also conducted. Our work is conducive to a better application of TCMs in complementary and alternative therapies in RA and OA.


Guidelines on the Use of Sex and Gender in Cardiovascular Research.
In cardiovascular research, sex and gender have not typically been considered in research design and reporting until recently. This has resulted in clinical research findings from which not only all women, but also gender-diverse individuals have been excluded. The resulting dearth of data has led to a lack of sex and gender specific clinical guidelines and raises serious questions about evidence-based care. Basic research has also excluded considerations of sex. Including sex and/or gender as research variables not only has the potential to improve the health of society overall now, it also provides a foundation of knowledge on which to build future advances. The goal of this guidelines article is to provide advice on best practices to include sex and gender considerations in study design, as well as data collection, analysis, and interpretation to optimally establish rigor and reproducibility needed to inform clinical decision-making and improve outcomes. In cardiovascular physiology, incorporating sex and gender is a necessary component when optimally designing and executing research plans. The guidelines serve as the first guidance on how to include sex and gender in cardiovascular research. We provide here a beginning path towards achieving this goal and improve the ability of the research community to interpret results through a sex and gender lens to enable comparison across studies and laboratories, resulting in better health for all.


[Effect of myelopeptides on the synthesis of DNA and total protein in the cells of the lymphoid organs in the mouse].
Using double 3H-thymidine/14C-amino acid label, the influence of myelopeptides (MP) on chromosomal DNA and total protein (without histones) synthesis has been studied in vitro in mouse lymphoid organ cells. It has been shown that MP cause a decrease in DNA labelling and a parallel increase in protein labelling in bone marrow cells and have practically no effect on these two parameters in thymus, lymph node and spleen cells. It has been found that MP have only mitogenic effect on the above-mentioned organ cells, but in combination with 2-mercaptoethanol. The data obtained show that MP possess the properties of cell-differentiating factors.


Improved dextran sulfate-calcium chloride method for the removal of nonspecific inhibitors with modifications for nonspecific agglutinin removal in the rubella hemagglutination inhibition test.
The dextran sulfate precipitation method for removal of nonspecific inhibitors from sera to be tested in the rubella hemagglutination-inhibition test was modified by shortening the dextran sulfate-serum mixture incubation period from 2 hr to 30 min. A comparative study using both treatment times showed no significant difference in rubella antibody titers. Reducing the treatment time even further also gave comparable results, indicating the select 30-min incubation period is not marginal for undertreatment. It was shown that 41.5% of the 159 serums tested had nonspecific agglutinins at levels from 1:8 to as high as 1:64. In each specimen, all demonstrable nonspecific agglutinin was removed by adsorption with 10%, 0- to 2-day-old chick red blood cells rather than the usually recommended 50% adult chicken erythrocytes.


Respiratory Symptoms Among Domestic Waste Collectors.
Increasing amount of waste concurrently increases the risk of exposure to hazardous materials among waste collectors. Vigorous exertion in the field intensifies the abundance of contaminated inspirable droplets. If left undetected and untreated, it may provoke significant pernicious health effects and redundant burdens to employees and employers. The purpose of this study is to determine the prevalence of respiratory symptoms and their associated factors among domestic waste collectors. A cross-sectional study was conducted in Kota Kinabalu, Sabah between January and April 2020. Data were collected using a validated questionnaire which encompasses details about sociodemographic, health status, environment, and employment characteristics. A total of 290 waste collectors with a mean age of 40 (±9) years old were participated in the study. Most of them were from Kadazan-Dusun-Murut ethnic origin with educational background till secondary school. The average monthly income of the workers was USD 298.45 (±171.9) per month, and they had been in service for 11 (±9.04) years. Respiratory symptoms were seen in 21% of the workers. The identified significant risk factors were determined as underlying chronic diseases (OR=2.34; 95% CI=1.054, 5.219) and contact with pets (OR=1.87; 95% CI=1.004, 3.288). Respiratory symptoms are prevalent amidst domestic waste collectors and related to their health and field activities.


The expression of adipogenic genes is decreased in obesity and diabetes mellitus.
Obesity is strongly correlated with type 2 diabetes mellitus, a common disorder of glucose and lipid metabolism. Although adipocytes are critical in obesity, their role in diabetes has only recently been appreciated. We conducted studies by using DNA microarrays to identify differences in gene expression in adipose tissue from lean, obese, and obese-diabetic mice. The expression level of over 11,000 transcripts was analyzed, and 214 transcripts showed significant differences between lean and obese mice. Surprisingly, the expression of genes normally associated with adipocyte differentiation were down-regulated in obesity. Not all obese individuals will become diabetic; many remain normoglycemic despite profound obesity. Understanding the transition to obesity with concomitant diabetes will provide important clues to the pathogenesis of type 2 diabetes. Therefore, we examined the levels of gene expression in adipose tissue from five groups of obese mice with varying degrees of hyperglycemia, and we identified 88 genes whose expression strongly correlated with diabetes severity. This group included many genes that are known to be involved in signal transduction and energy metabolism as well as genes not previously examined in the context of diabetes. Our data show that a decrease in expression of genes normally involved in adipogenesis is associated with obesity, and we further identify genes important for subsequent development of type 2 diabetes mellitus.


Serum transaminases and isocitric dehydrogenase in kwashiorkor.
Serum glutamic oxalacetic transaminase (G-OT), glutamic-pyruvic transaminase (G-PT), and isocitric dehydrogenase (ICD) have been estimated in six groups of subjects, namely, normal British adults, normal Nigerian adults, undernourished Nigerian adults and children, Nigerian children with florid protein malnutrition (kwashiorkor), and cases of protein malnutrition after treatment. The normal range in Nigerian adults was apparently slightly lower than in European adults.A marked increase in serum enzyme activity was found in florid malnutrition, and a slight rise in undernutrition. In all cases serum G-PT showed lesser changes than G-OT and ICD. The causes of these abnormalities are discussed. The increased serum enzyme activity is thought to be derived primarily from the liver and possibly also from voluntary muscle. Estimation of serum enzymes is recommended in the investigation of protein malnutrition.


The Effects of Viewing Cute Pictures on Performance During a Basketball Free-Throw Task.
Previous studies have shown that viewing cute pictures leads to performance improvement in a subsequent fine motor task. We examined the beneficial effects of viewing cute pictures in a more complex sporting skill (i.e., basketball free throws) by comparing three conditions (viewing baby animal pictures, adult animal pictures, and no pictures) and two tests (no-pressure and pressure). The participants, all of whom were college basketball players, performed 16 free throws in each condition. In the no-pressure test, male participants improved performance after viewing pictures of baby animals but not after adult animals and no pictures. In the pressure test, no significant improvement was observed. For female participants, the cuteness-viewing effect was not observed in both tests. The results suggest that viewing cute pictures may improve performance during basketball free throws in a low-pressure situation by narrowing the breadth of attentional focus and inducing approach motivation and caregiving behaviors.


New insights into the utilization of medium-chain triglycerides by the neonate: observations from a piglet model.
Because of their unique digestive and metabolic properties, medium-chain triglycerides (MCT) are used in a variety of nutritional settings, including use as a readily digestible energy source for the neonate. This review examines recent findings from our laboratory related to MCT digestion and metabolism that are drawn from a neonatal piglet model, but which may be clinically relevant to human infants. We have shown that MCT utilization improves rapidly with postnatal age (within 24 h), which is likely due to the ontogeny of pancreatic lipase. Additional data delineate the dramatic effects of emulsification and fatty acid chain length (within the medium-chain family) on utilization, with the suggestion that triacylhexanoate is utilized at the highest rate. Again, these effects are likely mediated via an increase in the kinetics of digestion rather than metabolism. Indeed, using both in vitro and in vivo radiotracer techniques, we were unable to detect metabolic differences among even-chain fatty acid homologues. However, studies with isolated hepatocytes have shown greater oxidation rates of odd-chain fatty acids compared with even-chain homologues, in part as a result of the anaplerotic potential of propionyl-CoA arising from odd-carbon fatty acid oxidation. In vivo radiotracer studies also showed an improvement in octanoate oxidation to CO2, with a concomitant reduction in urinary dicarboxylic acid excretion when colostrum-deprived piglets were supplemented with L-carnitine. Further metabolic research led to the novel finding that piglets have a very limited hepatic capacity to synthesize ketone bodies, and that acetate may be a relatively important product of hepatic fatty acid oxidation in this species.


Beyond detoxification: a role for mouse mEH in the hepatic metabolism of endogenous lipids.
Microsomal and soluble epoxide hydrolase (mEH and sEH) fulfill apparently distinct roles: Whereas mEH detoxifies xenobiotics, sEH hydrolyzes fatty acid (FA) signaling molecules and is thus implicated in a variety of physiological functions. These epoxy FAs comprise epoxyeicosatrienoic acids (EETs) and epoxy-octadecenoic acids (EpOMEs), which are formed by CYP epoxygenases from arachidonic acid (AA) and linoleic acid, respectively, and then are hydrolyzed to their respective diols, the so-called DHETs and DiHOMEs. Although EETs and EpOMEs are also substrates for mEH, its role in lipid signaling is considered minor due to lower abundance and activity relative to sEH. Surprisingly, we found that in plasma from mEH KO mice, hydrolysis rates for 8,9-EET and 9,10-EpOME were reduced by 50% compared to WT plasma. This strongly suggests that mEH contributes substantially to the turnover of these FA epoxides-despite kinetic parameters being in favor of sEH. Given the crucial role of liver in controlling plasma diol levels, we next studied the capacity of sEH and mEH KO liver microsomes to synthesize DHETs with varying concentrations of AA (1-30 μM) and NADPH. mEH-generated DHET levels were similar to the ones generated by sEH, when AA concentrations were low (1 μM) or epoxygenase activity was curbed by modulating NADPH. With increasing AA concentrations sEH became more dominant and with 30 μM AA produced twice the level of DHETs compared to mEH. Immunohistochemistry of C57BL/6 liver slices further revealed that mEH expression was more widespread than sEH expression. mEH immunoreactivity was detected in hepatocytes, Kupffer cells, endothelial cells, and bile duct epithelial cells, while sEH immunoreactivity was confined to hepatocytes and bile duct epithelial cells. Finally, transcriptome analysis of WT, mEH KO, and sEH KO liver was carried out to discern transcriptional changes associated with the loss of EH genes along the CYP-epoxygenase-EH axis. We found several prominent dysregulations occurring in a parallel manner in both KO livers: (a) gene expression of Ephx1 (encoding for mEH protein) was increased 1.35-fold in sEH KO, while expression of Ephx2 (encoding for sEH protein) was increased 1.4-fold in mEH KO liver; (b) Cyp2c genes, encoding for the predominant epoxygenases in mouse liver, were mostly dysregulated in the same manner in both sEH and mEH KO mice, showing that loss of either EH has a similar impact. Taken together, mEH appears to play a leading role in the hydrolysis of 8,9-EET and 9,10-EpOME and also contributes to the hydrolysis of other FA epoxides. It probably profits from its high affinity for FA epoxides under non-saturating conditions and its close physical proximity to CYP epoxygenases, and compensates its lower abundance by a more widespread expression, being the only EH present in several sEH-lacking cell types.


Dietary supplementation with arginine and glutamic acid modifies growth performance, carcass traits, and meat quality in growing-finishing pigs.
Sixty Duroc × Large White × Landrace pigs with an average initial BW of 77.1 ± 1.3 kg were used to investigate the effects of dietary supplementation with arginine and glutamic acid on growth performance, carcass traits, and meat quality in growing-finishing pigs. The animals were randomly assigned to 1 of 5 treatment groups (12 pigs/group, male:female ratio 1:1). The pigs in the control group were fed a basal diet (basal diet group), and those in the experimental groups were fed the basal diet supplemented with 2.05% -alanine (isonitrogenous group), 1.0% -arginine (Arg group), 1% glutamic acid + 1.44% -alanine (Glu group), or 1.0% -arginine + 1.0% glutamic acid (Arg+Glu group). After a 60-d period of supplementation, growth performance, carcass traits, and meat quality were evaluated. The results showed no significant differences ( > 0.05) in growth performance and carcass traits of the pigs in the Arg group relative to the basal diet group; however, the longissimus dorsi (LD) muscle and back fat showed a decrease ( < 0.05) in the percentage of SFA. In the Glu group, the final BW, phase 1 (d 1 to 30) and phase 2 (d 31 to 60) ADFI, and average back fat thickness of the pigs decreased ( < 0.05) by 7.14%, 23.43%, 8.03%, and 33.88%, respectively, when compared with the basal diet group. Dietary Arg+Glu supplementation had no effect ( > 0.05) on the final BW, phase 2 ADFI, and average daily weight gain in pigs but decreased ( < 0.05) their phase 1 ADFI, average back fat thickness, and percentage of SFA in the LD muscle and back fat, and increased ( < 0.05) the i.m. fat (IMF) content of the LD and biceps femoris muscles when compared with the basal diet group. Furthermore, a 16% decrease in yellowness (b* value; < 0.05) was observed in the Arg+Glu group pigs when compared with the isonitrogenous group. These findings suggest that dietary supplementation with both Arg and Glu beneficially increases the IMF deposition and improves the meat color and fatty acid composition without affecting growth performance and s.c. fat in pigs, providing a novel strategy to enhance meat quality in growing-finishing pigs.


The product of the leu-3 cistron as a regulatory element for the production of the leucine biosynthetic enzymes of Neurospora.
A class of intracistronic (or closely linked) partial reversions of leu-3 mutations has been found to be conditionally constitutive with respect to the synthesis of isopropylmalate isomerase (specified by the leu-2 cistron) and beta-isopropylmalate dehydrogenase (specified by the leu-1 cistron), two of the enzymes of leucine biosynthesis in Neurospora. The intermediate level of enzyme production by these leu-3(cc) mutants is independent of the obligatory inducer effector, alpha-isopropylmalate, but dependent upon the presence of the branched-chain amino acids, isoleucine, valine and leucine. The properties of leu-3+, leu-3 and leu-3(cc) in heterokaryons indicate that the transnuclear regulatory activity of the leu-3 product varies specifically as a function of available effector molecules. The information presented suggests that the leu-3 cistron is responsible not only for the production of a "positive" regulatory substance necessary for the expression of the leu-1 and leu-2 cistrons, but that it probably serves also a coordinating role in the expression of many of the genes involved in branched-chain amino acid metabolism.


CD56+ hematological neoplasms presenting in the skin: a retrospective analysis of 23 new cases and 130 cases from the literature.
The aim of this study was to define prognostic parameters and guidelines for diagnosis and treatment for CD56+ hematological neoplasms with first presentation in the skin. The study group included 153 cases (23 new and 130 from the literature). According to the World Health Organization classification, the group included 15 nasal and 38 nasal-type natural killer (NK)/T-cell lymphomas, 63 blastic NK-cell lymphomas, 14 cutaneous CD30+ lymphoproliferations, 10 cases of myeloid leukemia, six cases of subcutaneous panniculitis-like T-cell lymphoma (SCPLTCL) and seven peripheral T-cell lymphomas, unspecified. In general, these CD56+ hematological neoplasms had a poor prognosis, with only 27% of patients alive after a median follow-up of 12 months. The median survival was 13 months. Nasal and nasal-type NK/T-cell lymphomas and CD56+ SCPLTCL had the worst prognosis, with a median survival of 5, 6 and 5 months, respectively. Only nasal-type NK/T-cell lymphomas presenting with only skin lesions had a somewhat better prognosis (median survival 27 months). In blastic NK-cell lymphomas (median survival 14 months), age </=40 years, aggressive treatment with acute leukemia protocols and high TdT expression were associated with a more favorable prognosis. Striking similarities in histology, immunophenotype, clinical presentation and clinical behavior were found between blastic NK-cell lymphomas and CD56+ myeloid leukemias. CD56+ hematological neoplasms presenting in the skin have a poor prognosis, except for primary cutaneous CD30+ lymphoproliferations. The striking similarities between blastic NK-cell lymphomas and CD56+ myeloid leukemias presenting in the skin provide a rationale to treat these patients with more aggressive regimens, rather than with CHOP(-like) regimens and radiotherapy, which have proven to be inadequate therapies for this neoplasm.


An arrested late endosome-lysosome intermediate aggregate observed in a Chinese hamster ovary cell mutant isolated by novel three-step screening.
Chinese hamster ovary cell mutants defective in the post-uptake degradation of low-density lipoprotein (LDL) in lysosomes were selected from mutagenized cells by novel three-step screening. First, in the presence of LDL, clones sensitive to an inhibitor of the rate-limiting enzyme of the cholesterol biosynthetic pathway, 3-hydroxy-3-methylglutaryl-CoA reductase, were isolated. Second, from the selected clones, those lacking in the degradation of a constituent of a fluorescent LDL were qualitatively screened by microscopy. Third, the clones were further screened by previously established quantitative analytical flow cytometry that detects the early-phase disintegration of LDL by lysosomal acid hydrolases. One of the isolated mutant clones, LEX1 (Lysosome-Endosome X 1), was a recessive mutant, and exhibited a specific disorder in the late endocytic pathway. LEX1 cells showed an unusual perinuclear aggregate of vesicles, heterogeneously positive for lysosomal glycoprotein-B/cathepsin D and rab7, yet negative for the cation-independent mannose 6-phosphate receptor. The aggregate was formed around the microtubule organizing center, and was disrupted by nocodazole treatment. Internalized octadecyl rhodamine B-labeled LDL (R18-LDL) was accumulated in the perinuclear rab7-positive vesicles. In a Percoll density gradient, neither internalized R18-LDL nor internalized horseradish peroxidase was efficiently chased into heavy lysosomal fractions positive for beta-hexosaminidase. LEX1 cells showed differences in the activity and subcellular distribution of lysosomal enzymes. These characteristics of LEX1 cells are consistent with the ideas that the perinuclear vesicle aggregate is an arrested intermediate of direct fusion or divergence between lysosomes and rab7-positive, cation-independent mannose 6-phosphate receptor-negative late endosomes, and that equilibrium between the lysosomes and the late endosomes is shifted towards the late endosomes in LEX1 cells. Such fusion or divergence between the late endosomes and the lysosomes would determine an appropriate equilibrium between them, and might thereby play an important role for proper lysosomal digestive functions. LEX1 mutant cells would be helpful for the dissection of the as yet unrevealed details of the late endocytic membrane dynamics and for the identification of factors involved in the process arrested by the mutation.


High resolution allelotype of nonfunctional pancreatic endocrine tumors: identification of two molecular subgroups with clinical implications.
A high resolution allelotype for nonfunctional pancreatic endocrine tumors (NF-PETs) has been generated by microsatellite analysis of DNA from 16 frozen cases, each probed with 394 markers. Two subgroups of NF-PETs were found. Seven cases showed frequent, large allelic deletions [loss of heterozygosity (LOH)] with an average fractional allelic loss (FAL) of 0.55, whereas nine cases showed a small number of random losses with a FAL of 0.15. Designated high or low FAL, respectively, these genetic phenotypes showed correlation with the ploidy status: high-FAL tumors were aneuploid, low-FAL were diploid. Chromosomes 6q and 11q showed LOH in >60% of cases. About 50% of cases had losses on 11p, 20q, and 21. Selected LOH analysis on an additional 16 paraffin-embedded NF-PETs confirmed the high frequency of 6q and 11q LOH. The allelotype of NF-PET is markedly different from that of either ductal or acinar tumors of the pancreas as well as from that of functional-PETs. Moreover, whereas deletions involving chromosome 11 also are a feature of functional-PETs, the involvement of chromosome 6q is characteristic of NF-PETs. Survival analysis showed that none of the single chromosomal alterations was associated with outcome, whereas ploidy status is an independent factor adding prognostic information to that furnished by the proliferative index measured by Ki-67 immunohistochemistry.


Expression of the Qa-2k phenotype encoded by the Q5k gene on the surface of tumor cells derived from H-2k mice.
Immunological and biochemical characteristics of the Qa-2 murine nonclassical histocompatibility class 1 antigen expressed on tumor cells derived from H-2k (Qa-2-) mice were studied. It was found that the Qa-2 antigen on normal H-2b lymphocytes reacted with Qa-2-specific monoclonal antibodies (mAbs) 34-1.2, 59 (both specific to the alpha 1/alpha 2 region) and 141-15.8 (specific to the alpha 3 domain), and the Qa-2 antigen on H-2k tumor cells (Qa-2k antigen) reacted with mAbs 59 and 141-15.8, but not with 34-1.2. The normal Qa-2 antigen was susceptible to treatment with phosphatidylinositol-specific phospholipase C, but the Qa-2k antigen was insensitive to it. By Northern hybridization, polymerase chain reaction (PCR) studies on cDNA, Southern hybridization, Western blotting, and nucleotide sequence analysis, the Q5k gene was identified as the gene encoding the Qa-2k antigen expressed on BW5147 lymphoma cells derived from a mouse of AKR strain (H-2k, Qa-2-). The nucleotide sequence of PCR-amplified BW5147 Q5k cDNA showed complete agreement with the reported sequence of exons 1-5 of the Q5k gene of C3H/He. It also showed complete deletion of the region corresponding to exons 6 and 7, and a very short coding region in exon 8, resulting in very short cytoplasmic domain of the product compared with regular class 1 antigens. These characteristics were expected from the reported Q5k genomic sequence. These results revealed that the Qa-2k antigen was distinct from the normal Qa-2 antigen expressed on H-2b lymphocytes although it cross-reacted with some Qa-2-specific mAbs.


A J-protein is an essential subunit of the presequence translocase-associated protein import motor of mitochondria.
Transport of preproteins into the mitochondrial matrix is mediated by the presequence translocase-associated motor (PAM). Three essential subunits of the motor are known: mitochondrial Hsp70 (mtHsp70); the peripheral membrane protein Tim44; and the nucleotide exchange factor Mge1. We have identified the fourth essential subunit of the PAM, an essential inner membrane protein of 18 kD with a J-domain that stimulates the ATPase activity of mtHsp70. The novel J-protein (encoded by PAM18/YLR008c/TIM14) is required for the interaction of mtHsp70 with Tim44 and protein translocation into the matrix. We conclude that the reaction cycle of the PAM of mitochondria involves an essential J-protein.


The use of functional performance tests by primary health-care providers to determine walking ability with and without awalking device in community-dwelling elderly.
Background: Primary health-care (PHC) providers are important for community involvement in health promotion and prevention efforts, particularly today that the number of elderly is increasing dramatically. The use of a standard practical measure would help in promoting the effectiveness of referral and monitoring processes. Objectives: To investigate the use of functional performance tests in terms of the appropriate cutoff point to determine walking ability with and without a walking device in community-dwelling elderly, and reliability of the tests when used by PHC providers. Methods: Community-dwelling people aged 65 years or older who walked with or without a walking device (n = 309) were interviewed and assessed for information related to the use of a walking device in daily living. Then, they were randomly assessed for their functional ability using the Timed Up and Go Test (TUG), Five Times Sit-to-Stand Test (FTSST), and the 10-Meter Walk Test (10MWT). PHC providers, including a physical therapist, village health volunteer, and a caregiver, assessed 30 participants' functional performances, to address rater reliability of the tests. Results: The findings suggested that outcomes of the tests (TUG < 12 s, FTSST < 15 s, and 10MWT > 0.8 m/s) can indicate the ability of walking without a walking device of the participants. These tests could be used by PHC providers, except for the FTSST by a caregiver. Conclusions: The findings offer a clear cutoff point for promoting the involvement of PHC providers and the standardization of a screening, monitoring, and referral process among many clinical and community settings.


Posterior lumbar interbody fusion using compressive bone graft with allograft and autograft in the pyogenic discitis.
This is a retrospective study. To evaluate the advantages and effects of posterior lumbar interbody fusion (PLIF) using allograft and posterior instrumentation in the lumbar pyogenic discitis, which are resistant to antibiotics. To present preliminary results of PLIF using a compressive bone graft with allograft and pedicle screw fixation in the lumbar pyogenic discitis. Fifteen patients who had lumbar pyogenic discitis were treated by posterior approach from May 2004 to July 2008. The mean follow-up duration was 27.2 ± 18.68 months. The standing radiographs of the lumbar spine and clinical results were compared and analyzed in order to assess the bony union, the changes in the distance between the two vertebral bodies and the changes in the lordotic angle formed between the fused bodies immediately after surgery and at the final follow-up. Fifteen solid unions at an average of 15.2 ± 3.5 weeks after operation. The mean preoperative lordotic angle of the affected segments was 14.3 ± 15.1°, compared to 20.3 ± 12.3° after surgery and 19.8 ± 15.2° at last follow-up. For the functional result according to the Kirkaldy-Willis criteria, the outcome was excellent in 9, good in 5, fair in 1, and there were no poor cases. The average visual analogue scale score was decreased from 7.4 before surgery to 3.4 at 2 weeks postoperative. The main advantage in the procedure of PLIF using compressive bone graft with allograft and post instrumentation is early ambulation. We believe that this is another good procedure for patients with poor general condition because a further autograft bone harvest is not required.


The preprocessing of retinal images for the detection of fluorescein leakage.
Images of the human retina are routinely used in clinical practice for the diagnosis and management of eye disease. Increased permeability of retinal blood vessels, which is a clinically significant feature, can be visualized with a process known as fluorescein angiography as leakage of fluorescence dye into the surrounding tissues. Analyses of such images can be quantified but significant degradation of images due to uneven illumination or occluded optical pathways is often incurred during image capture. We describe a procedure to restore fluorescein angiographic retinal images so that quantitative computation can be reliably performed. Analysis of the image acquisition system reveals that captured images are composed of two functions, one describing the true underlying image and the other the degradation incurred. These two functions are independent of one another and it is possible to estimate the degradation from an isolated captured image and restore it appropriately. Any leakage of fluorescein dye is then detected by analysing the restored angiographic sequence over time and finding areas of the image that do not have the usual decrease in fluorescence intensity.


HER3-targeted protein chimera forms endosomolytic capsomeres and self-assembles into stealth nucleocapsids for systemic tumor homing of RNA interference in vivo.
RNA interference represents a potent intervention for cancer treatment but requires a robust delivery agent for transporting gene-modulating molecules, such as small interfering RNAs (siRNAs). Although numerous molecular approaches for siRNA delivery are adequate in vitro, delivery to therapeutic targets in vivo is limited by payload integrity, cell targeting, efficient cell uptake, and membrane penetration. We constructed nonviral biomaterials to transport small nucleic acids to cell targets, including tumor cells, on the basis of the self-assembling and cell-penetrating activities of the adenovirus capsid penton base. Our recombinant penton base chimera contains polypeptide domains designed for noncovalent assembly with anionic molecules and tumor homing. Here, structural modeling, molecular dynamics simulations, and functional assays suggest that it forms pentameric units resembling viral capsomeres that assemble into larger capsid-like structures when combined with siRNA cargo. Pentamerization forms a barrel lined with charged residues mediating pH-responsive dissociation and exposing masked domains, providing insight on the endosomolytic mechanism. The therapeutic impact was examined on tumors expressing high levels of HER3/ErbB3 that are resistant to clinical inhibitors. Our findings suggest that our construct may utilize ligand mimicry to avoid host attack and target the siRNA to HER3+ tumors by forming multivalent capsid-like structures.


Ezrin Promotes Antigen Receptor Diversity during B Cell Development by Supporting Ig H Chain Variable Gene Recombination.
Genome-level rearrangements of Ig genes during B cell development are critical for generation of a diverse repertoire of BCRs that bind to a multitude of foreign Ags and some self Ags. Bone marrow B cell development involves a variety of cell-cell interactions, cell migration, and receptor signaling that likely benefit from the activity of membrane-cytoskeletal reorganizing proteins. However, the specific contribution of such proteins toward BCR repertoire diversification is poorly understood. Ezrin is a membrane-cytoskeletal linker protein that regulates mature B cell activation through spatial organization of the BCR. We employed next-generation sequencing to investigate whether Ezrin plays a role in IgH rearrangements and generation of BCR diversity in developing bone marrow B cells. BCR repertoire development occurred stochastically in B cell progenitors from both control and B cell conditional Ezrin-deficient mice. However, the loss of Ezrin resulted in fewer unique CDRs (CDR3s) in the BCRs and reduced Shannon entropy. Ezrin-deficient pre-B cells revealed similar utilization of joining (J) genes but significantly fewer variable (V) genes, thereby decreasing V-J combinatorial diversity. V-J junctional diversity, measured by CDR3 length and nucleotide additions and deletions, was not altered in Ezrin-deficient pre-B cells. Mechanistically, Ezrin-deficient cells showed a marked decrease in RAG1 gene expression, indicating a less efficient DNA recombination machinery. Overall, our results demonstrate that Ezrin shapes the BCR repertoire through combinatorial diversification.


Decreased bone mineral density in Costello syndrome.
Costello syndrome (CS) is a multisystemic disorder characterized by postnatal reduced growth, facial dysmorphism, cardiac defects, cognitive impairment, skin and musculo-skeletal anomalies, and predisposition to certain cancers. CS is caused by activating germline mutations in the HRAS proto-oncogene. Similar to what is observed in other RASopathies, CS causative HRAS mutations promote enhanced signal flow through the RAF-MEK-ERK and PI3K-AKT signaling cascades. While decreased bone mineralization has been documented in other RASopathies, such as neurofibromatosis type 1 and Noonan syndrome, systematic studies investigating bone mineral density (BMD) are lacking in CS. Dual-energy X-ray absorptiometry (DXA) was utilized to assess BMD and body composition (fat and fat-free mass) in a cohort of subjects with molecularly confirmed diagnosis of CS (n = 9) and age-matched control individuals (n = 29). Using general linear regression, subtotal body (total body less head), lumbar, femoral neck and femur BMD parameters were compared considering age, sex, body mass index (BMI) and Tanner stage. Blood and urine biomarkers of bone metabolism were also assessed. All individuals with CS showed significantly lower mean values of subtotal, lumbar and femoral neck BMD compared to the control group (p ≤ 0.01). Similarly, mean total body mass and fat-free mass parameters were lower among the CS patients than in controls (p < 0.01). Low 25-OH vitamin D concentration was documented in all individuals with CS, with values below the reference range in two patients. No significant correlation between vitamin D levels and BMD parameters was observed. CS belongs to a family of developmental disorders, the RASopathies, that share skeletal defects as a common feature. The present data provide evidence that, similar to what is recently seen in NF1 and NS, bone homeostasis is impaired in CS. The significant decrease in BMD and low levels of vitamin D documented in the present cohort, along with the risk for pathologic fractures reported in adult individuals with CS, testifies the requirement for a preventive treatment to alleviate evolutive complications resulting from dysregulated bone metabolism.


Effects of the subacute administration of nefiracetam on abnormal behavior in aged rats.
We investigated the effects of nefiracetam on learning and memory by the Morris water maze task and water-finding test, and on emotional behavior by forced swimming, hole-board and open-field tests in old male Kbl Wistar rats aged 90 and 108 weeks. In the water maze task, the acquisition of the task in aged rats was slower than that in young rats. Subacute administration of nefiracetam (1 and 3 mg/kg daily) for 24 days tended to shorten the goal latency to escape onto the platform in a dose-dependent manner in the retention test, conducted 24 days after acquisition training. Nefiracetam (1 mg/kg) administration for 49 days decreased the duration of immobility in aged rats in the forced swimming test. Locomotor activity in young rats during the dark period was significantly higher than that during the light period, while there was no difference in locomotor activity between the light and dark periods in aged rats, suggesting that locomotor activity during the dark period and nocturnal habits may be impaired in aged rats. Subacute administration of nefiracetam for 14 days significantly increased the locomotor activity during the dark, but not light, period in a dose-dependent manner. In addition, nefiracetam given for 38 days, significantly shortened the increased time elapsed before animals started exploring the environment in aged rats compared with young rats in the water-finding tests. These findings suggest that nefiracetam may improve the impaired nocturnal habits and some of emotional behavior in aged rats.


The rise of hyperabundant native generalists threatens both humans and nature.
In many disturbed terrestrial landscapes, a subset of native generalist vertebrates thrives. The population trends of these disturbance-tolerant species may be driven by multiple factors, including habitat preferences, foraging opportunities (including crop raiding or human refuse), lower mortality when their predators are persecuted (the 'human shield' effect) and reduced competition due to declines of disturbance-sensitive species. A pronounced elevation in the abundance of disturbance-tolerant wildlife can drive numerous cascading impacts on food webs, biodiversity, vegetation structure and people in coupled human-natural systems. There is also concern for increased risk of zoonotic disease transfer to humans and domestic animals from wildlife species with high pathogen loads as their abundance and proximity to humans increases. Here we use field data from 58 landscapes to document a supra-regional phenomenon of the hyperabundance and community dominance of Southeast Asian wild pigs and macaques. These two groups were chosen as prime candidates capable of reaching hyperabundance as they are edge adapted, with gregarious social structure, omnivorous diets, rapid reproduction and high tolerance to human proximity. Compared to intact interior forests, population densities in degraded forests were 148% and 87% higher for wild boar and macaques, respectively. In landscapes with >60% oil palm coverage, wild boar and pig-tailed macaque estimated abundances were 337% and 447% higher than landscapes with <1% oil palm coverage, respectively, suggesting marked demographic benefits accrued by crop raiding on calorie-rich food subsidies. There was extreme community dominance in forest landscapes with >20% oil palm cover where two pig and two macaque species accounted for >80% of independent camera trap detections, leaving <20% for the other 85 mammal species >1 kg considered. Establishing the population trends of pigs and macaques is imperative since they are linked to cascading impacts on the fauna and flora of local forest ecosystems, disease and human health, and economics (i.e., crop losses). The severity of potential negative cascading effects may motivate control efforts to achieve ecosystem integrity, human health and conservation objectives. Our review concludes that the rise of native generalists can be mediated by specific types of degradation, which influences the ecology and conservation of natural areas, creating both positive and detrimental impacts on intact ecosystems and human society.


Colorectal cancer screening knowledge, attitudes and behavioural intention among Indigenous Western Australians.
Indigenous Australians are significantly less likely to participate in colorectal cancer (CRC) screening compared to non-Indigenous people. This study aimed to identify important factors influencing the decision to undertake screening using Faecal Occult Blood Testing (FOBT) among Indigenous Australians. Very little evidence exists to guide interventions and programmatic approaches for facilitating screening uptake in this population in order to reduce the disparity in colorectal cancer outcomes. Interviewer-administered surveys were carried out with a convenience sample (n = 93) of Indigenous Western Australians between November 2009-March 2010 to assess knowledge, awareness, attitudes and behavioural intent in regard to CRC and CRC screening. Awareness and knowledge of CRC and screening were low, although both were significantly associated with exposure to media advertising (p = 0.008; p < 0.0001). Nearly two-thirds (63%; 58/92) of respondents reported intending to participate in screening, while a greater proportion (84%; 77/92) said they would participate on a doctor's recommendation. Multivariate analysis with logistic regression demonstrated that independent predictors of screening intention were, greater perceived self-efficacy (OR = 19.8, 95% CI = 5.5-71.8), a history of cancer screening participation (OR = 6.8, 95% CI = 2.0-23.3) and being aged 45 years or more (OR = 4.5, 95% CI = 1.2-16.5). A higher CRC knowledge score (medium vs. low: OR = 9.9, 95% CI = 2.4-41.3; high vs. low: 13.6, 95% CI = 3.4-54.0) and being married or in a de-facto relationship (OR = 6.9, 95% CI = 2.1-22.5) were also identified as predictors of intention to screen with FOBT. Improving CRC related knowledge and confidence to carry out the FOBT self-screening test through education and greater promotion of screening has the potential to enhance Indigenous participation in CRC screening. These findings should guide the development of interventions to encourage screening uptake and reduce bowel cancer related deaths among Indigenous Australians.


VA-086 methacrylate gelatine photopolymerizable hydrogels: A parametric study for highly biocompatible 3D cell embedding.
The ability to replicate in vitro the native extracellular matrix (ECM) features and to control the three-dimensional (3D) cell organization plays a fundamental role in obtaining functional engineered bioconstructs. In tissue engineering (TE) applications, hydrogels have been successfully implied as biomatrices for 3D cell embedding, exhibiting high similarities to the natural ECM and holding easily tunable mechanical properties. In the present study, we characterized a promising photocrosslinking process to generate cell-laden methacrylate gelatin (GelMA) hydrogels in the presence of VA-086 photoinitiator using a ultraviolet LED source. We investigated the influence of prepolymer concentration and light irradiance on mechanical and biomimetic properties of resulting hydrogels. In details, the increasing of gelatin concentration resulted in enhanced rheological properties and shorter polymerization time. We then defined and validated a reliable photopolymerization protocol for cell embedding (1.5% VA-086, LED 2 mW/cm2) within GelMA hydrogels, which demonstrated to support bone marrow stromal cells viability when cultured up to 7 days. Moreover, we showed how different mechanical properties, derived from different crosslinking parameters, strongly influence cell behavior. In conclusion, this protocol can be considered a versatile tool to obtain biocompatible cell-laden hydrogels with properties easily adaptable for different TE applications.


Season is associated with Pseudomonas aeruginosa acquisition in young children with cystic fibrosis.
Pseudomonas aeruginosa, the principal respiratory pathogen in cystic fibrosis (CF) patients, is ubiquitous in the environment. Initial P. aeruginosa isolates in CF patients are generally environmental in nature. However, little information regarding seasonality of P. aeruginosa acquisition is available. We conducted a retrospective study to evaluate the seasonality of initial P. aeruginosa acquisition in young children with CF in the USA using the Cystic Fibrosis Foundation National Patient Registry from 2003 to 2009. Additionally, we assessed whether seasonal acquisition varied by climate zone. A total of 4123 children met inclusion criteria and 45% (n = 1866) acquired P. aeruginosa during a mean 2.0 years (SD 0.2 years) of follow up. Compared with winter, increased P. aeruginosa acquisition was observed in summer (incidence rate ratio (IRR): 1.22; 95% CI: 1.07-1.40) and autumn (IRR: 1.34; 95% CI: 1.18-1.52), with lower acquisition observed in spring (IRR: 0.81; 95% CI: 0.70-0.94). Seasonal variations in P. aeruginosa acquisition rates in the temperate and continental climate zones were similar to those in the overall cohort. In contrast, no significant seasonal effect was observed in the dry climate zone. In a corresponding analysis, no seasonal difference was observed in the rate of acquisition of Staphylococcus aureus, another common CF respiratory pathogen. These results provide preliminary support that climatic factors may be associated with initial P. aeruginosa acquisition in CF patients. Investigation and identification of specific risk factors, as well as awareness of seasonal variation, could potentially inform clinical recommendations including increased awareness of infection control and prevention strategies.


Identification of an aggression-promoting pheromone and its receptor neurons in Drosophila.
Aggression is regulated by pheromones in many animal species. However, in no system have aggression pheromones, their cognate receptors and corresponding sensory neurons been identified. Here we show that 11-cis-vaccenyl acetate (cVA), a male-specific volatile pheromone, robustly promotes male-male aggression in the vinegar fly Drosophila melanogaster. The aggression-promoting effect of synthetic cVA requires olfactory sensory neurons (OSNs) expressing the receptor Or67d, as well as the receptor itself. Activation of Or67d-expressing OSNs, either by genetic manipulation of their excitability or by exposure to male pheromones in the absence of other classes of OSNs, is sufficient to promote aggression. High densities of male flies can promote aggression by the release of volatile cVA. In turn, cVA-promoted aggression can promote male fly dispersal from a food resource, in a manner dependent on Or67d-expressing OSNs. These data indicate that cVA may mediate negative-feedback control of male population density, through its effect on aggression. Identification of a pheromone-OSN pair controlling aggression in a genetic organism opens the way to unravelling the neurobiology of this evolutionarily conserved behaviour.


Insights into Lantibiotic Immunity Provided by Bioengineering of LtnI.
The lantibiotic lacticin 3147 has been the focus of much research due to its broad spectrum of activity against many microbial targets, including drug-resistant pathogens. In order to protect itself, a lacticin 3147 producer must possess a cognate immunity mechanism. Lacticin 3147 immunity is provided by an ABC transporter, LtnFE, and a dedicated immunity protein, LtnI, both of which are capable of independently providing a degree of protection. In the study described here, we carried out an in-depth investigation of LtnI structure-function relationships through the creation of a series of fusion proteins and LtnI determinants that have been the subject of random and site-directed mutagenesis. We establish that LtnI is a transmembrane protein that contains a number of individual residues and regions, such as those between amino acids 20 and 27 and amino acids 76 and 83, which are essential for LtnI function. Finally, as a consequence of the screening of a bank of 28,000 strains producing different LtnI derivatives, we identified one variant (LtnI I81V) that provides enhanced protection. To our knowledge, this is the first report of a lantibiotic immunity protein with enhanced functionality.


The quality of dental faculty work-life: report on the 2007 dental school faculty work environment survey.
This report is the third in a series of articles on the dental school work environment commissioned by the American Dental Education Association's Commission on Change and Innovation in Dental Education. The report is based on the most extensive research to date on faculty satisfaction in the dental school environment. The purpose of the study was to assess faculty perceptions and recommendations related to work environment, sources of job satisfaction and dissatisfaction, and professional development needs. More broadly, the study intends to provide insight into the "change readiness" of dental schools to move forward with curricular improvements and innovations. Findings are based on 1,748 responses from forty-nine U.S. dental schools obtained during the time frame of February to April 2007. The total number of respondents constituted 17 percent of all U.S. dental school faculty. The average response rate per school was thirty-six (21 percent). To elucidate the data in terms of issues related to the quality of faculty work-life based on demographics, the authors compared perceptions of various aspects of the work culture in academic dentistry among faculty with different academic ranks and academic degrees and by other variables such as age and gender, tenure versus non-tenure appointments, and full- versus part-time status. Quantitative and qualitative analyses show that the majority of faculty members described themselves as very satisfied to satisfied with their dental school overall and with their department as a place to work. Tenured associate professors expressed the greatest level of dissatisfaction. Opportunities for and support of professional development emerged as an area requiring substantially more attention from dental schools. The authors of the study suggest that dental school leaders use these findings to assess their individual dental school's work environment and to plan changes as needed.


Controllable coating and reshaping of gold nanorods with tetracyanoquinodimethane.
Various nanoparticle surface layers allow unique functionalities. We developed a coating method with tetracyanoquinodimethane that forms solid layers through π stacking on gold nanorod surfaces. Its reaction mechanism was investigated with reaction time, aging time and surfactant concentration. Our method could be generalizable to different nanoparticle shapes and crystal facets.


The critical need for postmarketing surveillance in gene therapy for haemophilia.
The goal of gene therapy for haemophilia is to alter the clinical phenotype to a milder form or even cure, by increasing endogenous coagulation factor levels through transfer of a functional gene encoding the respective deficient coagulation factor and subsequent transgene expression. Over the past decade, there has been tremendous progress in gene therapy, particularly in use of liver-directed adeno-associated viral vectors, such that several programmes for both haemophilia A and B are in phase 3 trials. With regulatory approval of the first gene therapy product expected as early as mid-2020, there is an urgent need for a mechanism to collect long-term data on safety and variability and durability of efficacy. There will be elements required by regulators for postmarketing surveillance and additional data needed to enhance our understanding of gene therapy outcomes and their impact on the lives of people with haemophilia. The aim of this manuscript was to describe efforts underway by the American Thrombosis and Hemostasis Network and the World Federation of Hemophilia to collect long-term harmonized data and considerations of the European and US regulatory agencies, which will inform ongoing data collection. The status of data collection around gene therapy in haemophilia and important outcome measures were obtained by literature review. Each author described elements relevant to the activities of their organization. Support of all stakeholders in gene therapy, providers, patients, industry and regulators, augers successful capture of uniform long-term safety and efficacy data to ensure optimal treatment of people with haemophilia.


Meckel's diverticulum: value of ectopic tissue.
Cases of symptomatic Meckel's diverticulum treated surgically on an emergency basis during the last decade are reviewed. A series of 18 patients were divided into two groups depending on the presence or absence of ectopic tissue in the diverticulum. Group 1 consisted of five patients (28 percent) without ectopic tissue, and Group 2 consisted of 13 patients (72 percent) with ectopic tissue (8 gastric ectopia, 2 pancreatic ectopia, and 3 both gastric and pancreatic ectopia). Previous clinical records related to Meckel's diverticulum were found for 54 percent of the patients in Group 2 and for none of those in Group 1. The most common acute manifestations were intestinal occlusion (seven patients), digestive hemorrhage (five patients), and peritonitis (three patients). The postoperative course was 7 days in Group 1 and 15 days in Group 2 regardless of the surgical technique used. We conclude that the presence of ectopic tissue in patients with Meckel's diverticulum seems to be the main risk for occurrence of an acute nonmechanical complication. This complication appears more frequently and with more severity in young patients.


Optimization of the All-D Peptide D3 for Aβ Oligomer Elimination.
The aggregation of amyloid-β (Aβ) is postulated to be the crucial event in Alzheimer's disease (AD). In particular, small neurotoxic Aβ oligomers are considered to be responsible for the development and progression of AD. Therefore, elimination of thesis oligomers represents a potential causal therapy of AD. Starting from the well-characterized d-enantiomeric peptide D3, we identified D3 derivatives that bind monomeric Aβ. The underlying hypothesis is that ligands bind monomeric Aβ and stabilize these species within the various equilibria with Aβ assemblies, leading ultimately to the elimination of Aβ oligomers. One of the hereby identified d-peptides, DB3, and a head-to-tail tandem of DB3, DB3DB3, were studied in detail. Both peptides were found to: (i) inhibit the formation of Thioflavin T-positive fibrils; (ii) bind to Aβ monomers with micromolar affinities; (iii) eliminate Aβ oligomers; (iv) reduce Aβ-induced cytotoxicity; and (v) disassemble preformed Aβ aggregates. The beneficial effects of DB3 were improved by DB3DB3, which showed highly enhanced efficacy. Our approach yielded Aβ monomer-stabilizing ligands that can be investigated as a suitable therapeutic strategy against AD.


Family adjustment of deployed and non-deployed mothers in families with a parent deployed to Iraq or Afghanistan.
Almost nothing is known about the family and individual adjustment of military mothers who have deployed to the conflicts in Iraq or Afghanistan (Operations Iraqi and Enduring Freedom, and Operation New Dawn; OIF, OEF, OND), constituting a gap in psychologists' knowledge about how best to help this population. We report baseline data on maternal, child, parenting, and couple adjustment for mothers in 181 families in which a parent deployed to OIF/OEF/OND. Among this sample, 34 mothers had deployed at least once, and 147 mothers had experienced the deployment of a male spouse/partner. Mothers completed self-report questionnaires assessing past year adverse life events, war experiences (for deployed mothers only), posttraumatic stress disorder (PTSD) and depression symptoms, difficulties in emotion regulation, parenting, couple adjustment, and child functioning. Mothers who had deployed reported greater distress than non-deployed mothers (higher scores on measures of PTSD and depression symptoms), and slightly more past year adverse events. A moderate number of war experiences (combat and post-battle aftermath events) were reported, consistent with previous studies of women in current and prior conflicts. However, no differences were found between the two groups on measures of couple adjustment, parenting, or child functioning. Results are discussed in terms of the dearth of knowledge about deployed mothers, and implications for psychologists serving military families.


Cardiac rehabilitation is associated with reduced long-term mortality in patients undergoing combined heart valve and CABG surgery.
No reports have been published to date on the impact of cardiac rehabilitation (CR) on mortality in patients undergoing combined heart valve and coronary artery bypass graft (CABG) surgery (V + CABG), a procedure that has increased significantly in frequency in recent years. We identified consecutive patients who underwent V + CABG surgery in the Olmsted County from 1996 to 2007. Propensity scores were developed using more than 40 clinical, operative, and post-operative characteristics. The impact of CR on long-term mortality was assessed via landmark analysis and using propensity score regression adjustment and stratification techniques. A total of 201 patients were included in our study, in whom 86 deaths occurred over a mean follow up of 6.8 years. Forty-seven per cent of patients participated in CR, with a significant trend towards increased participation in recent years (p = 0.04). Conditional on 6-month survival and controlling for propensity factors as well as mortality risk factors, CR participation was associated with a significant reduction in mortality (propensity score adjustment: HR 0.48, p = 0.009; propensity score stratification: HR 0.48, p = 0.016). The absolute risk reduction over 10 years was 14.5% (number needed to treat = 7). Results did not differ significantly based on age, gender, emergent status, or history of heart failure or arrhythmias, but CR participation was more beneficial for patients who underwent a mitral valve procedure (HR 0.24, 95% CI 0.08-0.77). This is the first study reporting a significant survival benefit with CR participation in patients who have undergone combined V + CABG surgery. These findings provide evidence in support of recommendations for CR participation after V + CABG surgery.


The C. elegans HP1 homologue HPL-2 and the LIN-13 zinc finger protein form a complex implicated in vulval development.
HP1 proteins are essential components of heterochromatin and contribute to the transcriptional repression of euchromatic genes via the recruitment to specific promoters by corepressor proteins including TIF1 and Rb. The Caenorhabditis elegans HP1 homologue HPL-2 acts in the "synMuv" (synthetic multivulval) pathway, which defines redundant negative regulators of a Ras signaling cascade required for vulval induction. Several synMuv genes encode for chromatin-associated proteins involved in transcriptional regulation, including Rb and components of the Mi-2/NuRD and TIP60/NuA4 chromatin remodeling complexes. Here, we show that HPL-2 physically interacts in vitro and in vivo with the multiple zinc finger protein LIN-13, another member of the synMuv pathway. A variant of the conserved PXVXL motif found in many HP1-interacting proteins mediates LIN-13 binding to the CSD of HPL-2. We further show by in vivo localization studies that LIN-13 is required for HPL-2 recruitment in nuclear foci. Our data suggest that the LIN-13/HPL-2 complex may physically link a subset of the Rb related synMuv proteins to chromatin.


Delivering Coordinated Cancer Care by Building Transactive Memory in a Team of Teams.
Cancer care delivery is highly complex. Treatment involves coordination within oncology health-care teams and across other teams of referring primary and specialty providers (a team of teams). Each team interfaces with patients and caregivers to offer component parts of comprehensive care. Because patients frequently obtain specialty care from divergent health-care systems resulting in cross-system health-care use, oncology teams need mechanisms to coordinate and collaborate within and across health-care systems to optimize clinical outcomes for all cancer patients. Transactive memory is one potential strategy that can help improve comprehensive patient care delivery. Transactive memory is a process by which two or more team professionals develop a shared system for encoding, storing, and retrieving information. Each professional is responsible for retaining only part of the total information. Applying this concept to a team of teams results in system benefits wherein all teams share an understanding of specialized knowledge held by each component team. The patient's role as the unifying member of the team of teams is central to successful treatment delivery. This clinical case presents a patient who is receiving oral treatment for advanced prostate cancer within two health systems. The case emphasizes the potential for error when multiple teams function without a point team (the team coordinating efforts of all other primary and specialty teams) and when the specialty knowledge of providers and patients is not well integrated into all phases of the care delivery process.


Validation, Optimal Threshold Determination, and Clinical Utility of the Infant Risk of Overweight Checklist for Early Prevention of Child Overweight.
Previous research has demonstrated the predictive validity of the Infant Risk of Overweight Checklist (IROC). This study further establishes the predictive accuracy of the IROC using data from the Avon Longitudinal Study of Parents and Children (ALSPAC) and examines the optimal threshold for determining high risk of childhood overweight. Using the IROC algorithm, we calculated the risk of being overweight, based on International Obesity Task Force criteria, in the first year of life for 980 children in the ALSPAC cohort at 5 years. Discrimination was assessed by the area under the receiver operating curve (AUC c-statistic). Net reclassification index (NRI) was calculated for risk thresholds ranging from 2.5% to 30%, which determine cutoffs for identifying infants at risk of becoming overweight. At 5 years of age, 12.3% of boys and 19.6% of girls were categorized overweight. Discrimination (AUC c-statistic) ranged from 0.67 (95% confidence interval [CI], 0.62-0.72) when risk scores were calculated directly to 0.93 (95% CI, 0.88-0.98) when the algorithm was recalibrated and missing values of the risk factor algorithm were imputed. The NRI showed that there were positive gains in reclassification using risk thresholds from 5% to 20%, with the maximum NRI being at 10%. This study confirms that the IROC has moderately good validity for assessing overweight risk in infants and offers an optimal threshold for determining high risk. The IROC algorithm has been imbedded into a computer program for Proactive Assessment of Obesity Risk during Infancy, which facilitates early overweight prevention through communication of risk to parents.


Two-hit mechanism of cardiac arrhythmias in diabetic hyperglycaemia: reduced repolarization reserve, neurohormonal stimulation, and heart failure exacerbate susceptibility.
Diabetic hyperglycaemia is associated with increased arrhythmia risk. We aimed to investigate whether hyperglycaemia alone can be accountable for arrhythmias or whether it requires the presence of additional pathological factors. Action potentials (APs) and arrhythmogenic spontaneous diastolic activities were measured in isolated murine ventricular, rabbit atrial, and ventricular myocytes acutely exposed to high glucose. Acute hyperglycaemia increased the short-term variability (STV) of action potential duration (APD), enhanced delayed afterdepolarizations, and the inducibility of APD alternans during tachypacing in both murine and rabbit atrial and ventricular myocytes. Hyperglycaemia also prolonged APD in mice and rabbit atrial cells but not in rabbit ventricular myocytes. However, rabbit ventricular APD was more strongly depressed by block of late Na+ current (INaL) during hyperglycaemia, consistent with elevated INaL in hyperglycaemia. All the above proarrhythmic glucose effects were Ca2+-dependent and abolished by CaMKII inhibition. Importantly, when the repolarization reserve was reduced by pharmacological inhibition of K+ channels (either Ito, IKr, IKs, or IK1) or hypokalaemia, acute hyperglycaemia further prolonged APD and further increased STV and alternans in rabbit ventricular myocytes. Likewise, when rabbit ventricular myocytes were pretreated with isoproterenol or angiotensin II, hyperglycaemia significantly prolonged APD, increased STV and promoted alternans. Moreover, acute hyperglycaemia markedly prolonged APD and further enhanced STV in failing rabbit ventricular myocytes. We conclude that even though hyperglycaemia alone can enhance cellular proarrhythmic mechanisms, a second hit which reduces the repolarization reserve or stimulates G protein-coupled receptor signalling greatly exacerbates cardiac arrhythmogenesis in diabetic hyperglycaemia.


Legume-rhizobium symbiotic promiscuity and effectiveness do not affect plant invasiveness.
The ability to fix atmospheric nitrogen is thought to play an important role in the invasion success of legumes. Interactions between legumes and nitrogen-fixing bacteria (rhizobia) span a continuum of specialization, and promiscuous legumes are thought to have higher chances of forming effective symbioses in novel ranges. Using Australian Acacia species in South Africa, it was hypothesized that widespread and highly invasive species will be more generalist in their rhizobial symbiotic requirements and more effective in fixing atmospheric nitrogen compared with localized and less invasive species. To test these hypotheses, eight localized and 11 widespread acacias were examined using next-generation sequencing data for the nodulation gene, nodC , to compare the identity, species richness, diversity and compositional similarity of rhizobia associated with these acacias. Stable isotope analysis was also used to determine levels of nitrogen obtained from the atmosphere via symbiotic nitrogen fixation. No differences were found in richness, diversity and community composition between localized and widespread acacias. Similarly, widespread and localized acacias did not differ in their ability to fix atmospheric nitrogen. However, for some species by site comparisons, significant differences in δ15N isotopic signatures were found, indicating differential symbiotic effectiveness between these species at specific localities. Overall, the results support recent findings that root nodule rhizobial diversity and community composition do not differ between acacias that vary in their invasiveness. Differential invasiveness of acacias in South Africa is probably linked to attributes such as differences in propagule pressure, reasons for (e.g. forestry vs. ornamental) and extent of, plantings in the country.


Integrated pathway clusters with coherent biological themes for target prioritisation.
Prioritising candidate genes for further experimental characterisation is an essential, yet challenging task in biomedical research. One way of achieving this goal is to identify specific biological themes that are enriched within the gene set of interest to obtain insights into the biological phenomena under study. Biological pathway data have been particularly useful in identifying functional associations of genes and/or gene sets. However, biological pathway information as compiled in varied repositories often differs in scope and content, preventing a more effective and comprehensive characterisation of gene sets. Here we describe a new approach to constructing biologically coherent gene sets from pathway data in major public repositories and employing them for functional analysis of large gene sets. We first revealed significant overlaps in gene content between different pathways and then defined a clustering method based on the shared gene content and the similarity of gene overlap patterns. We established the biological relevance of the constructed pathway clusters using independent quantitative measures and we finally demonstrated the effectiveness of the constructed pathway clusters in comparative functional enrichment analysis of gene sets associated with diverse human diseases gathered from the literature. The pathway clusters and gene mappings have been integrated into the TargetMine data warehouse and are likely to provide a concise, manageable and biologically relevant means of functional analysis of gene sets and to facilitate candidate gene prioritisation.


Issues in Ethics in Orthopaedic Surgery Following the COVID-19 Pandemic.
The COVID-19 pandemic has caused changes in health care as well as human suffering, and consideration of the principles of ethics can build a foundation to consider dilemmas that have arisen. Diversity, equity, and inclusion have become key issues. Simulation training and the related ethics of its application have taken on new meaning. Access to health care continues to evolve and will need further evaluation in the years following the COVID-19 pandemic.


Positive correlation between adenosine deaminase activity and superoxide formation during phagocytosis.
Phagocytosis and membrane perturbation in mouse macrophages results in an increased superoxide ion production which is in direct proportion to the concomitant increase in adenosine deaminase activity. Since adenosine deaminase activity controls the amount of substrate available to xanthine oxidase, and the latter produces superoxide during turnover of its substrates, it is proposed that the purine salvage pathway is an important source of the superoxide requirement of macrophages. It is further proposed that this may be the basis, at least in part, for the mechanism of the association of immunodeficiency with adenosine deaminase deficiency.


Lack of Oestrogen Receptor Expression in Breast Cancer Cells Does Not Correlate with Kisspeptin Signalling and Migration.
Kisspeptin is an anti-metastatic mediator in many cancer types, acting through its receptor, KISS1R. However, controversy remains regarding its role in breast cancer since both pro- and anti-metastatic roles have been ascribed to it. In KISS1R overexpressing triple-negative breast cancer (TNBC) cells, stimulation has been associated with increased invasion and MMP-9 expression, leading to the suggestion that hormone receptor status determines the metastatic effects of kisspeptin. To assess the veracity of this claim, we compared endogenous KISS1R signalling and physiological output in the hormone receptor-negative MDA-MB-231 and BT-20 cell lines after KP-10 (shortest active kisspeptin peptide) stimulation. MDA-MB-231 cells are metastatic when implanted in mice while BT-20 are not and remain epithelial-like. We show that both cell lines express KISS1R mRNA and respond to KP-10 by elevating calcium mobilisation. However, KP-10 stimulation induced migration of MDA-MB-231, but not BT-20 cells, in a calcium-dependent manner. Moreover, only BT-20 cells responded to KP-10 by increasing ERK phosphorylation in a β-arrestin-dependent manner. Interestingly, both cell lines displayed different complements of β-arrestin 1 and 2 expression. Overall, our data shows that, in TNBC, it is not universally true that kisspeptin/KISS1R stimulate migration or pro-metastatic behaviour, as divergent responses were observed in the two TNBC lines tested. Whether this divergence is related to the observed differences in β-arrestin complements warrants further investigation and may enable further stratification of the ability of kisspeptin to influence breast tumour behaviour.


MRI Findings Suggestive of Metastatic Axillary Lymph Nodes in Patients with Invasive Breast Cancer.
This study aimed to investigate the diagnostic performance of features suggestive of nodal metastasis on preoperative MRI in patients with invasive breast cancer. We retrospectively reviewed the preoperative breast MRI of 192 consecutive patients with surgically proven invasive breast cancer. We analyzed MRI findings of axillary lymph nodes with regard to the size, long/short ratio, cortical thickness, shape and margin of the cortex, loss of hilum, asymmetry, signal intensity (SI) on T2-weighted images (T2WI), degree of enhancement in the early phase, and enhancement kinetics. Receiver operating characteristic (ROC) analysis, chi-square test, t test, and McNemar's test were used for statistical analysis. Increased shorter diameter, uneven cortical shape, increased cortical thickness, loss of hilum, asymmetry, irregular cortical margin, and low SI on T2WI were significantly suggestive of metastasis. ROC analysis revealed the cutoff value for the shorter diameter and cortical thickness as 8.05 mm and 2.75 mm, respectively. Increased cortical thickness (> 2.75 mm) and uneven cortical shape showed significantly higher sensitivity than other findings in McNemar's test. Irregular cortical margins showed the highest specificity (100%). Cortical thickness > 2.75 mm and uneven cortical shape are more sensitive parameters than other findings, and an irregular cortical margin is the most specific parameter for predicting axillary metastasis in patients with invasive breast cancer.


Nested case-control study of autoimmune disease in an asbestos-exposed population.
To explore the potential association between asbestos exposure and risk of autoimmune disease, we conducted a case-control study among a cohort of 7,307 current and former residents of Libby, Montana, a community with historical occupational and environmental exposure to asbestos-contaminated vermiculite. Cases were defined as those who reported having one of three systemic autoimmune diseases (SAIDs): systemic lupus erythematosus, scleroderma, or rheumatoid arthritis (RA). Controls were randomly selected at a 3:1 ratio from among the remaining 6,813 screening participants using frequency-matched age and sex groupings. The odds ratios (ORs) and 95% confidence intervals (CIs) for SAIDs among those >or=65 years of age who had worked for the vermiculite mining company were 2.14 (95% CI, 0.90-5.10) for all SAIDs and 3.23 (95% CI, 1.31-7.96) for RA. In this age group, exposure to asbestos while in the military was also an independent risk factor, resulting in a tripling in risk. Other measures of occupational exposure to vermiculite indicated 54% and 65% increased risk for SAIDs and RA, respectively. Those who had reported frequent contact with vermiculite through various exposure pathways also demonstrated elevated risk for SAIDs and RA. We found increasing risk estimates for SAIDs with increasing numbers of reported vermiculite exposure pathways (p<0.001). These preliminary findings support the hypothesis that asbestos exposure is associated with autoimmune disease. Refined measurements of asbestos exposure and SAID status among this cohort will help to further clarify the relationship between these variables.


Serosurvey of selected reproductive pathogens in domestic ruminants from Upper Egypt.
Toxoplasmosis, neosporosis, and Q fever are among the most important abortifacient diseases in ruminants worldwide. These diseases result in huge economic losses in livestock besides the fact that some of are of public health concern. The present study aimed to update the data about the current seroepidemiological situation of these diseases in Upper Egypt. A total of 411 blood samples were collected from small and large ruminants and serologically tested against the presence of T. gondii, N. caninum, and C. burnetii. Generalized estimating equation (GEE) models were performed to assess the potential risk factors associated with the exposure to these pathogens. The overall seroprevalence of T. gondii was 47.9% (197/411) with an individual seropositivity of 59.4% (63/106), 58.6% (17/29), 38.8% (54/139) and 46% (63/137) in cattle, buffalo, sheep and goats, respectively. Meanwhile, 9.7% (38/411) of the examined animals were tested positive for anti-N. caninum antibodies, with an individual seropositivity of 13.2% (12/106), 34.5% (10/29), 8.6% (12/139) and 2.9% (4/137) in cattle, buffalo, sheep and goats, respectively. Furthermore, the overall prevalence of antibodies against C. burnetii was 17.3% (63/411), and exposure to this pathogen was detected in 4.7% (5/106) of cattle, 19.3% (20/129) of sheep, 29.2% (38/130) of goats but none of the examined buffalo were found to be seropositive. A total of 12.1% (50/411) of the examined animals showed co-exposure to at least two of the tested pathogens. Regarding the potential risk factors, there were statistically significant differences among species in the frequency of exposure to the three tested pathogens. Age (> 6 months) was also shown to be a significant risk factor associated with T. gondii exposure. The results obtained provided updated information about the occurrence of three of the main reproductive pathogens in Upper Egypt. The high seropositivity values found for the tested zoonotic pathogens in most of the analyzed ruminant species suggest the necessity of performing additional in-depth studies to evaluate the epidemiology of these pathogens in the study area.


Natural history of sigmoid colon cancer: report of a patient observed for 4 years.
We report a case of sigmoid colon cancer that was left untreated for a period of 4 years, because the patient declined treatment. A 59-year-old man was found to have an early carcinoma of the sigmoid colon measuring approximately 12 mm in diameter. The lesion, initially a flat cancer, increased in height and became sessile 4 months later. Subsequently, the central portion of the lesion became ulcerated, leaving an elevated ring along its periphery. The lesion eventually evolved into an ulcerated, invasive cancer. This sequence has not been observed with colonoscopy before.


Effectiveness of pro-active organizational models in primary care for diabetes patients.
Demographic changes and chronicity are posing new challenges to health care systems. Our study aimed to examine how effectively the three different types of proactive primary care models adopted by three different regional health care systems in Italy were improving the quality of diabetes management by general practitioners. A coordinated Italian nationwide project to compare systematically the new proactive organizational models implemented at regional and local level (the MEDINA Project) involved several regions and their local health units (LHUs). A quasi-experimental study was conducted on a large dataset obtained by processing administrative databases. A combined indicator was developed to assess the quality of care delivered by primary care physicians, based on adherence to recommendations concerning patient monitoring and treatment. The study concerned 602 Italian general practitioners (GPs), 174 of them female, who were caring for a total of 753,366 patients (47,575 of them diabetic). Analyzing a total score, representing global adherence to a quality management of patients with diabetes, confirmed that GPs who had adopted the new model of care for their diabetic patients obtained better results than those who had not, so the new policy was generally effective. Our study showed that introducing new, proactive primary care models could sustain efforts made around the world to guarantee good-quality chronic disease management in the primary care setting.


[USJ Herbarium of Costa Rica: history and contributions].
In 2011 the Herbarium USJ of the University of Costa Rica became 80 years old and came up with 100 000 specimens of all the taxa that traditional botany studies. Data and figures on the history, the founders, and contributions of USJ to the knowledge of Costa Rican flora are summarized.


Prenatal exposure to imipramine alters early behavioral development and beta adrenergic receptors in rats.
Offspring of rats exposed to water or 15 mg/kg/dy of imipramine (IMI) on gestational day 8 through 20 were examined for behavioral and neurochemical development. IMI-treated mothers gained significantly less weight during pregnancy, but the percentage giving birth, length of gestation and litter size were unaffected. Body weights of IMI-exposed offspring were significantly lower than controls until postnatal day 14 and brain weights were lower until day 30. In IMI-exposed pups, eye-opening occurred significantly earlier, development of the surface righting reflex was delayed and development of negative geotaxis was altered. Hypothalamic levels of noradrenaline and adrenaline were unchanged in IMI pups at 7, 14 and 30 days, but dopamine levels, unaffected at 7 and 14 days, were significantly lower than controls at 30 days. The number of cortical beta adrenergic receptors, measured by [3H]dihydroalprenolol binding, was significantly decreased by 18.7% at 14 days and 9.1% at 30 days. Affinity for binding was increased at 30 days. Brain levels of IMI and its metabolite desmethylimipramine were detectable in newborns prenatally exposed to IMI and desmethylimipramine/IMI ratios were 1.8 times those found in adults. The results indicate that prenatal exposure to IMI produces behavioral and neurochemical consequences lasting well past cessation of drug exposure.


Molecular characterization of alpha-keratins in comparison to associated beta-proteins in soft-shelled and hard-shelled turtles produced during the process of epidermal differentiation.
The tough corneous layer in the carapace and plastron of hard-shelled turtles derives from the accumulation of keratin-associated beta-proteins (KAbetaPs, formerly called beta-keratins) while these proteins are believed to be absent in soft-shelled turtles. Our bioinformatics and molecular study has instead shown that the epidermis of the soft-shelled turtle Apalone spinifera expresses beta-proteins like or even in higher amount than in the hard-shelled turtle Pseudemys nelsoni. The analysis of a carapace cDNAs library has allowed the identification and characterization of three alpha-keratins of type I and of ten beta-proteins (beta-keratins). The acidic alpha-keratins probably combine with the basic beta-proteins but the high production of beta-proteins in A. spinifera is not prevalent over that of alpha-keratin so that their combination does not determine the formation of hard corneous material. Furthermore the presence of a proline and cisteine in the beta-sheet region of beta-proteins in A. spinifera may be unsuited to form hard masses of corneous material. The higher amount of beta-proteins over alpha-keratins instead occurs in keratinocytes of the hard and inflexible epidermis of P. nelsoni determining the deposition of hard corneous material. The study suggests that the hardness of the corneous layer derives not exclusively from the interactions between alpha-keratins with KAbetaPs but also from the different dynamic of accumulation and loss of corneocytes in the corneous layer of the hard shelled turtles where a prevalent accumulation and piling of corneocytes takes place versus the soft shelled turtle where a rapid turnover of the stratum corneum occurs.


Pillar[5]arene-based [1]rotaxane: high-yield synthesis, characterization and application in Knoevenagel reaction.
We report a quantitative synthetic strategy of a [1]rotaxane from a representative pseudo[1]rotaxane. The structure of the [1]rotaxane was characterized by 1H NMR, 13C NMR, 2D NMR, mass spectroscopy, and melting point, and its optimized geometry in CHCl3 by theoretical calculation at the B3LYP/6-31G(D) level using the PCM model matched well with 2D NOESY. This [1]rotaxane proves to be a good catalyst for the Knoevenagel reaction in CHCl3, which follows second order kinetics.


Elevated levels of CD4(+)CD25(+)FoxP3(+) T cells in systemic sclerosis patients contribute to the secretion of IL-17 and immunosuppression dysfunction.
Immune imbalance between regulatory T (Treg) and Th17 cells is a characteristic of systemic sclerosis (SSc). The functional heterogeneity among Treg can be elucidated by separating Treg into different subsets based on the expression of FoxP3 and CD45RA. The aim of this study was to investigate the role of Treg subsets in the immune imbalance in naïve SSc. Peripheral blood mononuclear cells (PBMCs) of 31 SSc patients and 33 healthy controls were analyzed for the expression of CD4, CD25, CD45RA, CTLA-4, FoxP3, and IL-17 using flow cytometry. Treg immunesuppression capacity was measured in co-culture experiments. The expression of FoxP3, CTLA-4, IL-17A, and RORC mRNA was measured by real-time PCR. The frequency of CD4(+)CD25(+)FoxP3(+) Treg cells was significantly elevated in patients with SSc (3.62±1.14 vs 1.97±0.75, p<0.001) with diminished immunosuppression capacity. In SSc, the proportion of FoxP3(high)CD45RA(-) activated Treg cells (aTreg) was decreased, the proportion of FoxP3(low)CD45RA(-) T cells was increased, and the proportion of FoxP3(low)CD45RA(+) resting Treg cells (rTreg) was decreased. The immune suppression capacity of aTreg and rTreg was diminished, while FoxP3(low)CD45RA(-) T cells exhibited a lack of suppression capacity. The immune dysfunction of aTreg was accompanied by the abnormal expression of CTLA-4. Th17 cell numbers were elevated in SSc, FoxP3(low)CD45RA(-) T cells produced IL-17, confirming their Th17 potential, which was consistent with the elevated levels of FoxP3(+)IL-17(+) cells in SSc. A decrease in aTreg levels, along with functional deficiency, and an increase in the proportion of FoxP3(low)CD45RA(-) T cells, was the reason for the increase in dysfunctional Treg in SSc patients, potentially causing the immune imbalance between Treg and Th17 cells.


Virtual Reality in Preoperative Planning of Complex Cranial Surgery.
Changing paradigms of neurosurgical training and limited operative exposure during the residency period have made it necessary to evaluate newer technologies for training. Virtual reality (VR) technology provides three-dimensional reconstruction of routine imaging, along with the ability to see as well as interact. The application of VR technology in operative planning, which is an important part of neurosurgical training, has been incompletely studied so far. Sixteen final-year residents, post-M.Ch. (magister chirurgiae) residents, and fellows were included as study participants. They were divided into 2 groups based on their seniority for further analysis. Five complex cranial cases were selected and a multiple-choice question-based test was prepared by the authors, with 5 questions for each of the cases. The pretest score was determined based on performance on the test after participants accessed routine preoperative imaging. The posttest score was calculated after use of the VR system (ImmersiveTouch VR System, ImmersiveTouch Inc.). Analysis was performed by the investigators, who were blinded to the identity of the participant. Subanalysis based on the type of case and type of question was performed. Feedback was obtained from each participant regarding VR use. There was an overall improvement in scores from pretest to posttest, which was also noted in the analysis based on the participants' seniority. This improvement was noted to be more for the vascular cases (15.89%) compared with the tumor cases (7.84%). Participants also fared better in questions related to surgical anatomy and surgical approach, compared with questions based on the diagnosis. There was overall positive feedback from participants regarding VR use, and most participants wanted VR to become a routine part of operative planning. Our study shows that there is improvement in understanding of surgical aspects after use of this VR system.


Physicians' Opinions on Anticoagulant Therapy in Patients with a Limited Life Expectancy.
Patients with a limited life expectancy have an increased risk of thromboembolic and bleeding complications. Anticoagulants are often continued until death, independent of their original indication. We aimed to identify the opinions of physicians about the use of anticoagulants at the end of life. A mixed-method research design was used. A secondary analysis was performed on data from a vignette study and an interview study. Participants included general practitioners and clinical specialists. Physicians varied in their opinions: some would continue and others would stop anticoagulants at the end of life because of the risk of thromboembolic or bleeding complications. The improvement or preservation of patients' quality of life was a reason for both stopping and continuing anticoagulants. Other factors considered in the decision-making were the types of anticoagulant, the indication for which the anticoagulant was prescribed, underlying diseases, and the condition and life expectancy of the patient. Factors that made decision-making difficult were the lack of evidence on either strategy, uncertainty about patients' life expectancy, and the fear of harming patients. Which decision was eventually made seems largely dependent on the choice of the patient. In conclusion, there is a substantial variation in physicians' opinions regarding the use of anticoagulants in patients with a limited life expectancy. Physicians agree that the primary goal of medical care at end of life is the improvement or preservation of patients' quality of life. An important barrier to decision-making is the lack of evidence about the risks and benefits of stopping anticoagulants.


Comparative 16S rRNA oligonucleotide analyses and murein types of round-spore-forming bacilli and non-spore-forming relatives.
The phylogenetic incoherency of the genus Bacillus as presently described is demonstrated by analysis of both published and new data from comparative 16S rRNA cataloguing of nine Bacillus species and a number of related non-Bacillus taxa, i.e. Caryophanon latum, Filibacter limicola and Planococcus citreus. While the ellipsoidal-spore-forming bacilli, e.g. B. subtilis and allied species, formed a coherent cluster, the round-spore-forming bacilli showed a higher degree of relationship to the non-spore-forming organisms than these bacilli show among each other. Thus B. sphaericus clustered with C. latum, B. globisporus grouped with F. limicola, B. pasteurii with Sporosarcina ureae, and 'B. aminovorans' with P. citreus, respectively. These organisms formed two related subclusters which, in their phylogenetic depth, are comparable to that of the B. subtilis subline. With the exception of 'B. aminovorans', the 16S rRNA phylogeny was entirely consistent with the distribution of murein types. Even more distantly related to and grouping outside the main Bacillus cluster was B. stearothermophilus, which displayed a moderate relationship to Thermoactinomyces vulgaris. Taxonomic problems arising from the new insights into the intrageneric relationships of Bacillus are discussed.


Fitness Consequences of Female Alternative Reproductive Tactics in House Mice (Mus musculus domesticus).
Alternative reproductive tactics (ARTs) are defined as discrete differences in morphological, physiological, and/or behavioral traits associated with reproduction that occur within the same sex and population. House mice provide a rare example of ARTs in females, which can rear their young either solitarily or together with one or several other females in a communal nest. We assessed the fitness consequences of communal and solitary breeding in a wild population to understand how the two tactics can be evolutionarily stable. Females switched between the two tactics (with more than 50% of all females having two or more litters using both tactics), pointing toward communal and solitary breeding being two tactics within a single strategy and not two genetically determined strategies. Communal breeding resulted in reduced pup survival and negatively impacted female reproductive success. Older and likely heavier females more often reared their litters solitarily, indicating that females use a condition-dependent strategy. Solitary breeding seems the more successful tactic, and only younger and likely less competitive females might opt for communal nursing, even at the cost of increased pup mortality. This study emphasizes the importance of analyzing phenotypic plasticity and its role in cooperation in the context of female ARTs.