Microbial inositol polyphosphate metabolic pathway as drug development target.
Inositol polyphosphates are a diverse and multifaceted class of intracellular messengers omnipresent in eukaryotic cells. These water-soluble molecules regulate many aspects of fundamental cell physiology. Removing this metabolic pathway is deleterious: inositol phosphate kinase null mutations can result in lethality or substantial growth phenotypes. Inositol polyphosphate synthesis occurs through the actions of a set of kinases that phosphorylate phospholipase-generated IP3 to higher phosphorylated forms, such as the fully phosphorylated IP6 and the inositol pyrophosphates IP7 and IP8. Unicellular organisms have a reduced array of the kinases for synthesis of higher phosphorylated inositol polyphosphates, while human cells possess two metabolic routes to IP6. The enzymes responsible for inositol polyphosphate synthesis have been identified in all eukaryote genomes, although their amino acid sequence homology is often barely detectable by common search algorithms. Homology between human and microbial inositol phosphate kinases is restricted to a few catalytically important residues. Recent studies of the inositol phosphate metabolic pathways in pathogenic fungi (Cryptococcus neoformans) and protozoa (Trypanosome brucei) have revealed the importance of the highly phosphorylated inositol polyphosphates to the fitness and thus virulence of these pathogens. Given this, identification of inositol kinase inhibitors specifically targeting the kinases of pathogenic microorganisms is desirable and achievable.


Aorto-iliac aneurysms rupturing into the iliac veins.
Rupture of aorto-iliac aneurysms into the iliac veins is seldom reported in distinct series, and represents only a fraction of ruptures into the abdominal cavity. Four cases are reported: two aortic aneurysms ruptured into the right iliac vein, one right iliac aneurysm ruptured into the right iliac vein, one right iliac aneurysm ruptured into the cava at the bifurcation, one aortoiliac aneurysm ruptured into the right iliac vein. Two patients showed severe shock at admission. Central venous pressure at operation was 43.6 +/- 26.3 cmH2O, and rapidly decreased at aortic clamping. Venous bleeding was controlled by finger compression of the iliac and caval vein and aortic graft replacement followed direct suture repair of the venous breach. De Weese caval clip was implanted twice, to prevent pulmonary embolism in severely compromised iliac veins. This series represents 1% among 373 ruptured abdominal aortic aneurysms operated on as emergencies over the same period. Pulmonary embolism, either paradoxical or postoperative, never occurred. Mortality rate was 25%, compared to 35% overall mortality among the whole ruptured group. The role of the site of rupture is swelling of the lower limbs is suggested, and edema can be suggestive though statistical evidence has not been achieved. Three concepts are stressed: early diagnosis and operation (i.e. before cardiac high output failure and shock); accurate as well as simple surgical technique; careful post-operative intensive care.


The magic of MAGI-1: A scaffolding protein with multi signalosomes and functional plasticity.
MAGI-1 is a critical cellular scaffolding protein with over 110 different cellular and microbial protein interactors. Since the discovery of MAGI-1 in 1997, MAGI-1 has been implicated in diverse cellular functions such as polarity, cell-cell communication, neurological processes, kidney function, and a host of diseases including cancer and microbial infection. Additionally, MAGI-1 has undergone nomenclature changes in response to the discovery of an additional PDZ domain, leading to lack of continuity in the literature. We address the nomenclature of MAGI-1 as well as summarize many of the critical functions of the known interactions. Given the importance of many of the interactors, such as human papillomavirus E6, the Coxsackievirus and adenovirus receptor (CAR), and PTEN, the enhancement or disruption of MAGI-based interactions has the potential to affect cellular functions that can potentially be harnessed as a therapeutic strategy for a variety of diseases.


The effect of hospital-based health promotion on the health practices of full-time hospital nurses: a cross-sectional study.
Many studies have reported positive contributions of health promotion on the health behavior of nursing staff working in hospitals, including the maintenance of a regular healthy diet, engagement in physical activity, performance of routine screening practices, and participation in a health examination. Despite being considered a role model for healthy lifestyles, little is known about the effect of health-promoting hospital settings on nursing staff. The aim of this study was to perform a nationwide, hospital-based, cross-sectional, survey comparing health practices between full-time nurses of health-promoting hospitals and those of non-health-promoting hospitals in Taiwan. We conducted a nationwide, hospital-based, cross-sectional, survey in 100 hospitals from May to July 2011 using a questionnaire as the measurement tool. Nurses aged between 18 and 65 years from certified health-promoting hospitals (n = 14,769) were compared with nurses in non-health-promoting hospitals (n = 11,242). A multiple logistic regression model was conducted to estimate the effect of certified HPH status on the likelihood of performing health behavior, receiving general physical examination, undergoing cancer screening, and participating in hospital-based health-promoting activities. All nurses of HPH hospitals were more likely to perform physical activity, practice cancer screening, receive at least one general physical examination in the past 3 years, and had a higher chance of participating in at least one hospital-based health-promoting activity in the past year (particularly weight-control groups and sports-related clubs) than those of non-HPH hospitals. This study suggests the effectiveness of implementing health promotion on the health behavior of full-time nursing staff in hospitals.


Spin-orbit-driven magnetic structure and excitation in the 5d pyrochlore Cd2Os2O7.
Much consideration has been given to the role of spin-orbit coupling (SOC) in 5d oxides, particularly on the formation of novel electronic states and manifested metal-insulator transitions (MITs). SOC plays a dominant role in 5d(5) iridates (Ir(4+)), undergoing MITs both concurrent (pyrochlores) and separated (perovskites) from the onset of magnetic order. However, the role of SOC for other 5d configurations is less clear. For example, 5d(3) (Os(5+)) systems are expected to have an orbital singlet with reduced effective SOC. The pyrochlore Cd2Os2O7 nonetheless exhibits a MIT entwined with magnetic order phenomenologically similar to pyrochlore iridates. Here, we resolve the magnetic structure in Cd2Os2O7 with neutron diffraction and then via resonant inelastic X-ray scattering determine the salient electronic and magnetic energy scales controlling the MIT. In particular, SOC plays a subtle role in creating the electronic ground state but drives the magnetic order and emergence of a multiple spin-flip magnetic excitation.


The effect of clomiphene citrate, herbal mixture, and herbal mixture along with clomiphene citrate on clinical and para-clinical parameters in infertile women with polycystic ovary syndrome: a randomized controlled clinical trial.
To evaluate the effect of a herbal mixture (i.e., Mentha spicata, Zingiber officinale, Cinnamomum zeylanicum, and Citrus sinensis) alone and in combination with clomiphene citrate (CC) compared to CC on the treatment of polycystic ovary syndrome (PCOS). This single-blind randomized clinical trial was conducted on 60 infertile participants with PCOS who were randomly divided into three groups. After spontaneous or progestin-induced withdrawal bleeding, group 1 (n = 20) received routine treatment with CC (50-150 mg) for three menstrual cycles from the 3rd to 5th day of menstruation for 5 days while group 2 (n = 20) and group 3 (n = 20) received herbal mixture capsules 700 mg/day and the herbal mixture along with CC for 3 months, respectively. Finally, several related parameters were measured, including the level of sex steroids, homeostatic model assessment for insulin resistance (HOMA-IR), lipid profile (primary outcomes), thyroid hormones, and clinical features. The analysis was based on intention-to-treat analysis. No statistically significant differences were observed between the groups in terms of socio-demographic characteristics. However, after adjustment for baseline, luteinizing hormone (aMD = 4.9; 95% CI: 3.7-6.2), luteinizing hormone/follicle-stimulating hormone (aMD = 0.9; 95% CI: 0.7-1.2), total testosterone (aMD = -0.12; 95% CI: -0.2 to -0.01) in group 2 and free testosterone (aMD = -6.0; 95% CI: -9.7 to -2.3) in group 3 revealed a significant difference compared to group 1. In addition, HOMA-IR in group 2 (aMD = -1.3; 95% CI: -2.4 to -0.2) decreased significantly compared to group 1. Further, total cholesterol, triglycerides, low-density lipoprotein cholesterol, and very low-density lipoprotein cholesterol decreased significantly in group 2 (aMD = -21.8; 95% CI: -31.5 to -12.1; aMD = -29.9; 95% CI: -47.9 to -12.0; aMD = -21.2; 95% CI: -31.3 to -11.1; aMD = -5.1; 95% CI: -7.5 to -2.7) and group 3 (aMD = -18.3; 95% CI: -27.4 to -9.2; aMD = -26.9; 95% CI: -43.8 to -9.9; aMD = -21.4; 95% CI: -31.1 to -11.7; aMD = -5.9; 95% CI: -8.3 to -3.6) compared to group 1, respectively. However, high-density lipoproteins cholesterol in group 2 (aMD = 6.8; 95% CI: 2.9-10.7) and group 3 (aMD = 10.7; 95% CI: 7.2-14.7) increased remarkably compared to group 1. Overall, clinical outcomes improved significantly in all groups (p < 0.05). In general, the herbal mixture along with CC was found to improve free testosterone, HOMA-IR, lipid profile, and clinical features of PCOS women.


Cell number regulator genes in Prunus provide candidate genes for the control of fruit size in sweet and sour cherry.
Striking increases in fruit size distinguish cultivated descendants from small-fruited wild progenitors for fleshy fruited species such as Solanum lycopersicum (tomato) and Prunus spp. (peach, cherry, plum, and apricot). The first fruit weight gene identified as a result of domestication and selection was the tomato FW2.2 gene. Members of the FW2.2 gene family in corn (Zea mays) have been named CNR (Cell Number Regulator) and two of them exert their effect on organ size by modulating cell number. Due to the critical roles of FW2.2/CNR genes in regulating cell number and organ size, this family provides an excellent source of candidates for fruit size genes in other domesticated species, such as those found in the Prunus genus. A total of 23 FW2.2/CNR family members were identified in the peach genome, spanning the eight Prunus chromosomes. Two of these CNRs were located within confidence intervals of major quantitative trait loci (QTL) previously discovered on linkage groups 2 and 6 in sweet cherry (Prunus avium), named PavCNR12 and PavCNR20, respectively. An analysis of haplotype, sequence, segregation and association with fruit size strongly supports a role of PavCNR12 in the sweet cherry linkage group 2 fruit size QTL, and this QTL is also likely present in sour cherry (P. cerasus). The finding that the increase in fleshy fruit size in both tomato and cherry associated with domestication may be due to changes in members of a common ancestral gene family supports the notion that similar phenotypic changes exhibited by independently domesticated taxa may have a common genetic basis.


Cognitive exploration drives engagement and re-engagement with imaginary worlds, but not spatial exploration as predicted by evolutionary theory.
The empirical evidence for exploration underlying the appeal of imaginary worlds is mostly absent or contradictory. Openness, and the cognitive exploration it represents, provides a better account than the overall drive to explore predicted by evolutionary theory. Furthermore, exploration cannot explain why imaginary worlds foster frequent re-engagement.


One-year MR imaging follow-up of patients with multiple sclerosis under cortisone therapy.
Twenty patients with relapsing/remitting course of MS were studied four times with MR imaging over the course of one year. First MR was undertaken during the acute relapse, afterwards patients were given cortisone therapy for four to six weeks. The second MR study followed 4-6 weeks after the first, the patients at this time being in remission. The third MR study was carried out 4 months after the first, the last scan one year after the first. The total number of lesions varied, though not greatly, over the whole follow-up, but there was an influence of the clinical course of MS on the pattern of lesions in MR imaging, mostly in respect to the number of confluences and the size of the lesions. Follow-up over one year showed that the inflammatory process produced an increase in the number of plaques, independent of the fact that most patients stayed in remission. A delayed effect of the cortisone therapy on the size, number, and confluence of plaques is suggested whilst clinical signs improved in most cases immediately after the beginning of drug therapy. Independent of the clinical course of the disease in some cases plaques previously seen vanished and others appeared in one and the same examination.


Change in self-rated general health is associated with perceived illness burden: a 1-year follow up of patients newly diagnosed with type 2 diabetes.
Diabetic patients' lifestyle adaptations to improve glycaemic control are not always followed by improvements in self-rated general health (SRH). The perceived impact of diabetes on patients' daily lives may influence changes in their SRH. This paper examines the association of illness severity, treatment, behavioural, and coping-related factors with changes in SRH from diagnosis of type 2 diabetes until one year later, in a population-based sample of 599 patients aged 40 years or over who were treated in general practice. Change in SRH was estimated by a cumulative probit model with the inclusion of covariates related to SRH (e.g. illness severity at diagnosis, behaviour, treatment, and the perceived impact of diabetes on patients' daily lives one year later). At diagnosis, 11.6% of patients reported very good, 35.1% good, 44.6% fair and 8.5% poor SRH. Physical inactivity, many diabetes-related symptoms, and cardiovascular disease were related to lower SRH ratings. On average SRH improved by 0.46 (95% CI: 0.37; 0.55) during the first year after diagnosis without inclusion of covariates. Mental and practical illness burden was the only factor associated with change in SRH, independent of patients' diabetes severity and medical treatment (p = 0.03, multivariate analysis). Compared to otherwise similar patients without illness burden, increase in SRH was marginally smaller among patients who expressed minor illness burden, but much smaller among patients with more pronounced illness burden. Much as one would expect, many patients increased their SRH during the first year after diabetes diagnosis. This increase in SRH was not associated with indicators of illness severity or factors reflecting socio-demographic circumstances, but patients experiencing illness burden had a smaller increase than those who reported no illness burden. We suggest that during the diabetes consultation, general practitioners explore further how patients manage their illness burden. We further suggest that diabetes guidelines extend their current focus on clinical and social aspects of diabetes to include questions on patient's perceived illness burden and SRH.


Deep learning based classification of facial dermatological disorders.
Common properties of dermatological diseases are mostly lesions with abnormal pattern and skin color (usually redness). Therefore, dermatology is one of the most appropriate areas in medicine for automated diagnosis from images using pattern recognition techniques to provide accurate, objective, early diagnosis and interventions. Also, automated techniques provide diagnosis without depending on location and time. In addition, the number of patients in dermatology departments and costs of dermatologist visits can be reduced. Therefore, in this work, an automated method is proposed to classify dermatological diseases from color digital photographs. Efficiency of the proposed approach is provided by 2 stages. In the 1st stage, lesions are detected and extracted by using a variational level set technique after noise reduction and intensity normalization steps. In the 2nd stage, lesions are classified using a pre-trained DenseNet201 architecture with an efficient loss function. In this study, five common facial dermatological diseases are handled since they also cause anxiety, depression and even suicide death. The main contributions provided by this work can be identified as follows: (i) A comprehensive survey about the state-of-the-art works on classifications of dermatological diseases using deep learning; (ii) A new fully automated lesion detection and segmentation based on level sets; (iii) A new adaptive, hybrid and non-symmetric loss function; (iv) Using a pre-trained DenseNet201 structure with the new loss function to classify skin lesions; (v) Comparative evaluations of ten convolutional networks for skin lesion classification. Experimental results indicate that the proposed approach can classify lesions with high performance (95.24% accuracy).


Prognostic Role of Human Epididymis Protein4 (HE4) in Endometrial Lesions: Study in a Tertiary Care Centre.
Endometrial cancer accounts for 20% of malignant tumours in the female reproductive system. A novel biological marker human epididymis protein4 (HE4) represents an important alternative indicator which may benefit patient mortality. To correlate the immunohistochemical expression of HE4 in different non-neoplastic and neoplastic endometrial lesions and with the WHO grade of the tumours. Our study was a cross-sectional, observational study done in a tertiary care hospital from December 2019 to June 2021 on the hysterectomy sample of 50 patients with a clinical history of abnormal uterine bleeding and pelvic pain. The study showed strong positivity of HE4 in cases of endometrial carcinoma, weak positivity in cases of atypical endometrial hyperplasia, and negativity in cases of endometrial hyperplasia without atypia group. WHO grade 3(50%) and grade 2 (29%) endometrioid adenocarcinoma NOS in our study showed strong positivity for HE4 which was statistically significant (P value = 0.001). In recent studies using overexpression of HE4-related genes, the malignant biological behaviour such as cell adhesion, invasion, and proliferation was enhanced. It was seen in our study that strong positivity of HE4 was seen in all endometrial carcinoma groups and with higher WHO grade. So, HE4 may become a potential therapeutic target for advanced-stage endometrial carcinoma which requires further research. Thus, human epididymis-specific protein 4 (HE4) has been shown to be a promising marker for the detection of endometrial carcinoma patients who could be benefitted from targeted therapy.


Radiomics performs comparable to morphologic assessment by expert radiologists for prediction of response to neoadjuvant chemoradiotherapy on baseline staging MRI in rectal cancer.
To compare the performance of advanced radiomics analysis to morphological assessment by expert radiologists to predict a good or complete response to chemoradiotherapy in rectal cancer using baseline staging MRI. We retrospectively assessed the primary staging MRIs [prior to chemoradiotherapy (CRT)] of 133 rectal cancer patients from 2 centers. First, two expert radiologists subjectively estimated the likelihood of achieving a "complete response" (ypT0) and "good response" (TRG 1-2), using a 5-point score (based on TN-stage, MRF/EMVI-status, size/signal/shape). Next, tumor volumes were segmented on high b value DWI (semi-automated, corrected by 2 non-expert and 2-expert readers, resulting in 5 segmentations), copied to the remaining sequences after which a total of 2505 radiomic features were extracted from T2W, low and high b value DWI and ADC. Stability of features for noise due to inter-reader and inter-scanner and protocol variations was assessed using intraclass correlation (ICC) and the Kruskal-Wallis test. Using data from center 1 (n = 86; training set), top 9 features were selected using minimum Redundancy Maximum Relevance and combined in a logistic regression model. Finally, diagnostic performance of the fitted models was assessed on data from center 2 (n = 47; validation set) and compared to the performance of the radiologists. The Radiomic models resulted in AUCs of 0.69-0.79 (with similar results for the segmentations performed by expert/non-expert readers) to predict response, results similar to the morphologic prediction by the expert radiologists (AUC 0.67-0.83). Radiomics using semi-automatically generated segmentations (without manual input) did not result in significant predictive performance. Radiomics could predict response to therapy with comparable diagnostic performance as expert radiologists, regardless of whether image segmentation was performed by non-expert or expert readers, indicating that expert input is not required in order for the radiomics workflow to produce significant predictive performance.


FEV1/FVC Severity Stages for Chronic Obstructive Pulmonary Disease.
Rationale: The diagnosis of chronic obstructive pulmonary disease (COPD) is based on a low FEV1/FVC ratio, but the severity of COPD is classified using FEV1% predicted (ppFEV1). Objectives: To test a new severity classification scheme for COPD using FEV1/FVC ratio, a more robust measure of airflow obstruction than ppFEV1. Methods: In COPDGene (Genetic Epidemiology of COPD) (N = 10,132), the severity of airflow obstruction was categorized by Global Initiative for Chronic Obstructive Lung Disease (GOLD) stages 1-4 (ppFEV1 of ⩾80%, ⩾50-80%, ⩾30-50%, and <30%). A new severity classification (STaging of Airflow obstruction by Ratio; STAR) was tested in COPDGene-FEV1/FVC ⩾0.60 to <0.70, ⩾0.50 to <0.60, ⩾0.40 to <0.50, and <0.40, respectively, for stages 1-4-and applied to the combined Pittsburgh SCCOR and Emphysema COPD Research Registry for replication (N = 2,017). Measurements and Main Results: The agreements (weighted Bangdiwala B values) between GOLD and the new FEV1/FVC ratio severity stages were 0.89 in COPDGene and 0.88 in the Pittsburgh cohort. In COPDGene and the Pittsburgh cohort, compared with GOLD staging, STAR provided significant discrimination between the absence of airflow obstruction and stage 1 for all-cause mortality, respiratory quality of life, dyspnea, airway wall thickness, exacerbations, and lung function decline. No major differences were noted for emphysema, small airway disease, and 6-minute-walk distance. The STAR classification system identified a greater number of adults with stage 3/4 disease who would be eligible for lung transplantation and lung volume reduction procedure evaluations. Conclusions: The new STAR severity classification scheme provides discrimination for mortality that is similar to the GOLD classification but with a more uniform gradation of disease severity. STAR differentiates patients' symptoms, disease burden, and prognosis better than the existing scheme based on ppFEV1, and is less sensitive to race/ethnicity and other demographic characteristics.


Primary Preventive Care of Hematopoietic Stem Cell Transplantation Survivors: Time to Educate and Empower Recipients and Providers.
Increasing use of hematopoietic stem cell transplantation (HCT) and improvements in recipient outcomes have led to a steady increase in the number of allogeneic HCT survivors. In addition to complications specific to the transplantation process, HCT recipients are at increased risk of developing cardiovascular disease (CVD) and subsequent neoplasm (SN). Strict surveillance of risk factors for CVD and cancer in the general population is recommended as an essential component of long-term follow-up (LTFU) care of HCT survivors, but implementation of this has been suboptimal. Various models for improving the provision of survivorship care have been proposed, including a hybrid/combined care approach wherein the HCT provider manages transplantation-specific complications and the primary care physician (PCP) provides general medical care, including surveillance and aggressive management of CVD risk factors and screening for SN. This model also offers a practical approach to LTFU care for HCT survivors who live at a distance from the HCT center, which is a reality for many recipients of HCT at The Ottawa Hospital (TOH). As the success of such a hybrid approach to survivorship care depends on the engagement of HCT recipients with their PCP and compliance with recommended general population surveillance, the aim of the present study was to assess the rates of PCP attendance and adherence to recommended preventive medicine interventions in the years immediately before and after HCT. We hypothesized that rates would be suboptimal and planned to use these results as a baseline for an educational initiative aimed at increasing awareness of HCT recipients and their PCPs about embracing preventive survivorship care. This was a single-center cohort study of allogeneic HCT recipients who underwent transplantation at TOH with linkage to population-based health administrative data. Published clinical practice guidelines were used to define recommended screening for CVD risk factors and cancer. The rates of annual PCP visits and utilization of recommended preventive care interventions in the 5 years before and after HCT were calculated for all eligible patients. Between 2014 and 2020, 409 patients with provincial health care coverage underwent allogeneic HCT at TOH. The median patient age was 51 years (range, 15 to 73 years), with a male predominance (60.9%). Approximately one-quarter of recipients did not attend a PCP visit in the 5 years before and after transplantation, and this proportion increased to one- third in the fifth year post-HCT. Among those recipients who were eligible, only 20% to 25% underwent recommended screening for dyslipidemia and diabetes. Cancer screening rates were also low, at 16% to 18% for cervical cancer, 18% to 22% for colon cancer, and 30% to 31% for breast cancer. These results highlight the need to increase awareness of HCT recipients and their PCPs about the risk of developing CVD and SN post-transplantation and to emphasize the potential to mitigate this risk by adhering to recommendations for surveillance to enable prompt intervention. Patient education should incorporate this information and empower HCT survivors to actively engage in their follow-up care and optimize their long-term outcomes.


A survey of tuberculosis infection control practices at the NIH/NIAID/DAIDS-supported clinical trial sites in low and middle income countries.
Health care associated transmission of Mycobacterium tuberculosis (TB) is well described. A previous survey of infection control (IC) practices at clinical research sites in low and middle income countries (LMIC) funded by the National Institute of Allergy and Infectious Diseases (NIAID) conducting HIV research identified issues with respiratory IC practices. A guideline for TB IC based on international recommendations was developed and promulgated. This paper reports on adherence to the guideline at sites conducting or planning to conduct TB studies with the intention of supporting improvement. A survey was developed that assessed IC activities in three domains: facility level measures, administrative control measures and environmental measures. An external site monitor visited each site in 2013-2014, to complete the audit. A central review committee evaluated the site-level survey and results were tabulated. Fisher's exact test was performed to determine whether there were significant differences in practices at sites that had IC officers versus sites that did not have IC officers. Significance was assessed at p</=.05 RESULTS: Seven of thirty-three sites surveyed (22 %) had all the evaluated tuberculosis IC (TB IC) elements in place. Sixty-one percent of sites had an IC officer tasked with developing and maintaining TB IC standard operating procedures. Twenty-two (71 %) sites promptly identified and segregated individuals with TB symptoms. Thirty (93 %) sites had a separate waiting area for patients, and 26 (81 %) collected sputum within a specific well-ventilated area that was separate from the general waiting area. Sites with an IC officer were more likely to have standard operating procedures covering TB IC practices (p = 0.02) and monitor those policies (p = 0.02) and perform regular surveillance of healthcare workers (p = 0.02). The presence of an IC officer had a positive impact on performance in most of the TB IC domains surveyed including having adequate ventilation (p = 0.02) and a separate area for sputum collection (p = 0.02) CONCLUSIONS: Specific and targeted support of TB IC activities in the clinical research environment is needed and is likely to have a positive and sustained impact on preventing the transmission of TB to both health care workers and vulnerable HIV-infected research participants.


A novel antimycin analogue antimycin A2c, derived from marine Streptomyces sp., suppresses HeLa cells via disrupting mitochondrial function and depleting HPV oncoproteins E6/E7.
Novel antimycin alkaloid antimycin A2c (AE) was isolated from the culture of a marine derived Streptomyces sp. THS-55. We elucidated its chemical structure by extensive spectra and clarified the specific mechanism in HPV infected-cervical cancer. Colony formation assay, cell cycle analysis, hoechst 33342 staining assay, et.al were used to detect the inhibitory effect of AE on cervical cancer cells. Meanwhile, flow cytometry, western blotting, immunoprecipitation, RNA interference and molecular docking were used to analyze the mechanism of AE. AE exhibited potent cytotoxicity in vitro against HPV-transformed cervical cancer HeLa cell line. AE inhibited the proliferation, arrested cell cycle distribution, and triggered caspase dependent apoptosis in HeLa cells. Further studies revealed AE-induced apoptosis is mediated by the degradation of E6/E7 oncoproteins. Molecular mechanic investigation showed that AE degraded the levels of E6/E7 oncoproteins through reactive oxygen (ROS)-mediated ubiquitin-dependent proteasome system activation, and the increased ROS generation was due to the disruption of the mitochondrial function. This present work revealed that this novel marine derived antimycin alkaloid could target the mitochondria and subsequently degrade HPV E6/E7 oncoproteins, and have potential application in the design and development of lead compound for cervical cancer cells, as well as the development for tool compounds to dissect E6/E7 functions.


Feedback from Arctic charr: Feed flavour stimulation and re-feeding after feed deprivation stimulate genes encoding both orexigenic and anorexigenic neuropeptides.
Despite vast research attention, the knowledge about central mechanisms of appetite regulation in teleost remains inconclusive. A common strategy in studies on appetite regulating mechanisms is to measure the response to feed restriction or - deprivation, but responses vary between fish species and between experiments, and are also likely dependent on the degree of energy perturbation. The anadromous Arctic charr is an interesting model for studying appetite regulation as its feeding cycle comprises months of winter anorexia, and hyperphagia during summer. Here we studied how the gene expression of putative hypothalamic appetite regulators were affected by two days, one week and one month feed deprivation during summer, and subsequent re-feeding and exposure to feed flavour. Short-term feed deprivation caused only a minor reduction in condition factor and had no effect on hypothalamic gene expression. Long-term feed-deprivation caused a marked reduction in weight and condition factor which contrasted the increase in weight and condition factor seen in ad libitum fed controls. A marked energy perturbation by feed deprivation was also indicated by a lower hypothalamic expression of the genes encoding insulin-like growth factor 1 (IGF1) and IGF1 binding protein 5 in the feed deprived charr compared to fed controls. Surprisingly, long-term feed deprivation and energy perturbation did not induce changes in hypothalamic appetite regulators. Unexpectedly, re-feeding and exposure to feed flavour caused an increase in the expression of the genes encoding the orexigenic agouti-related peptide and the anorexigenic melanocortin receptor 4 and cocaine- and amphetamine-regulated transcript. Our study gives strong evidence for a role of these in appetite regulation in Arctic charr, but their mechanisms of action remain unknown. We suggest that changes in gene expression are more likely to be registered during transition phases, e.g. from fasting to feeding and upon stimulatory inputs such as feed flavour.


Monitoring the growth dynamics of Tetragenococcus halophilus strains in lupine moromi fermentation using a multiplex-PCR system.
The microbiota of a seasoning sauce fermentation process is usually complex and includes multiple species and even various strains of one species. Moreover, composition and cell numbers of individual strains vary over the course of the entire fermentation. This study demonstrates the applicability of a multiplex PCR system to monitor growth dynamics of Tetragenococcus (T.) halophilus strains in order to evaluate their performance and help to select the most competitive starter strain. In a previous study we isolated T. halophilus strains from multiple lupine moromi fermentation processes and characterized them. In this study we wanted to monitor the growth dynamics of these strains in a competitive lupine moromi model fermentation process using a multiplex PCR system. Therefore, pasteurized lupine koji was inoculated with eight different T. halophilus strains, six from lupine moromi, one from an experimental buckwheat moromi fermentation process and the type strain DSM 20,339T, to create the inoculated lupine moromi pilot scale fermentation process. With the multiplex PCR system, we could detect that all strains could grow in lupine moromi but, that TMW 2.2254 and TMW 2.2264 outperformed all other strains. Both strains dominated the fermentation after three weeks with cell counts between 4 × 106 to 4 × 107 CFU/mL for TMW 2.2254 and 1 × 107 to 5 × 107 CFU/mL for TMW 2.2264. The pH dropped to value below 5 within the first 7 days, the selection of these strains might be related to their acid tolerance.


Influence of body weight on patients' satisfaction with ambulatory care.
Patients with obesity experience psychosocial consequences because of their weight and report physician bias. We examined whether obesity is associated with lower patient satisfaction with ambulatory care among 2,858 patients seen at 11 academically affiliated primary care practices in Boston. Compared with normal weight patients (body mass index [BMI], 19.0 to 24.9 kg/M 2), overweight (BMI, 25.0 to 29.9 kg/M 2) and obese patients (BMI > or =30 kg/M 2) reported lower overall satisfaction scores at their most recent visit; the scores were 85.5, 85.0, and 82.6 out a possible 100, respectively (P =.05). After adjustment for potential confounders including illness burden, obese patients reported lower scores but the difference was not statistically significant (mean difference, 1.23 [95% confidence interval -0.67 to 3.12]). Patient satisfaction with their usual provider and their practice did not vary by BMI group. Obesity is associated with only modest decreases in satisfaction scores with the most recent visit, which were explained largely by higher illness burden among obese patients.


High bacterial biodiversity increases degradation performance of hydrocarbons during bioremediation of contaminated harbor marine sediments.
We investigated changes of bacterial abundance and biodiversity during bioremediation experiments carried out on oxic and anoxic marine harbor sediments contaminated with hydrocarbons. Oxic sediments, supplied with inorganic nutrients, were incubated in aerobic conditions at 20 °C and 35 °C for 30 days, whereas anoxic sediments, amended with organic substrates, were incubated in anaerobic conditions at the same temperatures for 60 days. Results reported here indicate that temperature exerted the main effect on bacterial abundance, diversity and assemblage composition. At higher temperature bacterial diversity and evenness increased significantly in aerobic conditions, whilst decreased in anaerobic conditions. In both aerobic and anaerobic conditions, biodegradation efficiencies of hydrocarbons were significantly and positively related with bacterial richness and evenness. Overall results presented here suggest that bioremediation strategies, which can sustain high levels of bacterial diversity rather than the selection of specific taxa, may significantly increase the efficiency of hydrocarbon degradation in contaminated marine sediments.


YIPF5 (p.W218R) mutation induced primary microcephaly in rabbits.
Primary microcephaly (PMCPH) is a rare autosomal recessive neurodevelopmental disorder with a global prevalence of PMCPH ranging from 0.0013% to 0.15%. Recently, a homozygous missense mutation in YIPF5 (p.W218R) was identified as a causative mutation of severe microcephaly. In this study, we constructed a rabbit PMCPH model harboring YIPF5 (p.W218R) mutation using SpRY-ABEmax mediated base substitution, which precisely recapitulated the typical symptoms of human PMCPH. Compared with wild-type controls, the mutant rabbits exhibited stunted growth, reduced head circumference, altered motor ability, and decreased survival rates. Further investigation based on model rabbit elucidated that altered YIPF5 function in cortical neurons could lead to endoplasmic reticulum stress and neurodevelopmental disorders, interference of the generation of apical progenitors (APs), the first generation of progenitors in the developing cortex. Furthermore, these YIPF5-mutant rabbits support a correlation between unfolded protein responses (UPR) induced by endoplasmic reticulum stress (ERS), and the development of PMCPH, thus providing a new perspective on the role of YIPF5 in human brain development and a theoretical basis for the differential diagnosis and clinical treatment of PMCPH. To our knowledge, this is the first gene-edited rabbit model of PMCPH. The model better mimics the clinical features of human microcephaly than the traditional mouse models. Hence, it provides great potential for understanding the pathogenesis and developing novel diagnostic and therapeutic approaches for PMCPH.


The insular cortex: a review.
The human insular cortex forms a distinct, but entirely hidden lobe, situated in the depth of the Sylvian fissure. Here, we first review the recent literature on the connectivity and the functions of this structure. It appears that this small lobe, taking up less than 2% of the total cortical surface area, receives afferents from some sensory thalamic nuclei, is (mostly reciprocally) connected with the amygdala and with many limbic and association cortical areas, and is implicated in an astonishingly large number of widely different functions, ranging from pain perception and speech production to the processing of social emotions. Next, we embark on a long, adventurous journey through the voluminous literature on the structural organization of the insular cortex. This journey yielded the following take-home messages: (1) The meticulous, but mostly neglected publications of Rose (1928) and Brockhaus (1940) are still invaluable for our understanding of the architecture of the mammalian insular cortex. (2) The relation of the insular cortex to the adjacent claustrum is neither ontogenetical nor functional, but purely topographical. (3) The insular cortex has passed through a spectacular progressive differentiation during hominoid evolution, but the assumption of Craig (2009) that the human anterior insula has no homologue in the rhesus monkey is untenable. (4) The concept of Mesulam and Mufson (1985), that the primate insula is essentially composed of three concentrically arranged zones, agranular, dysgranular, and granular, is presumably correct, but there is at present much confusion concerning the more detailed architecture of the anterior insular cortex. (5) The large spindle-shaped cells in the fifth layer of the insular cortex, currently known as von Economo neurons (VENs), are not only confined to large-brained mammals, such as whales, elephants, apes, and humans, but also occur in monkeys and prosimians, as well as in the pygmy hippopotamus, the Atlantic walrus, and Florida manatee. Finally, we point out that the human insula presents a unique opportunity for performing an in-depth comparative analysis of the relations between structure and function in a typical sensory and a typical cognitive cortical domain.


Three-year outcome following neonatal encephalopathy in a high-survival cohort.
This study investigated the 3-year clinical outcomes in relation to the severity of encephalopathy in high-survival infants who underwent therapeutic hypothermia. This retrospective observational study was conducted in level II/III neonatal intensive care units in Japan. The nationwide cohort included 474 infants registered in the Baby Cooling Registry of Japan between January 2012 and December 2016. Clinical characteristics, mortality rate and severe neurological impairment at age 3 years were evaluated. Of the infants, 48 (10.4%), 291 (63.1%) and 122 (26.5%) had mild, moderate and severe encephalopathy, respectively, upon admission. By age 3, 53 (11.2%) infants died, whereas 110 (26.1%) developed major disabilities. The mild group survived up to age 3. In the moderate group, 13 (4.5%) died and 44 (15.8%) developed major disabilities. In the severe group, 39 (32.0%) died by age 3. Adverse outcomes were observed in 100 (82.0%) infants. Mortality was relatively low in all subgroups, but the incidence of major disabilities was relatively high in the severe group. The relatively low mortality and high morbidity may be due to Japanese social and ethical norms, which rarely encourage the withdrawal of intensive life support. Cultural and ethical backgrounds may need to be considered when assessing the effect of therapeutic interventions.


Increased Sperm DNA Fragmentation in Infertile Men with Varicocele: Relationship with Apoptosis, Seminal Oxidative Stress, and Spermatic Parameters.
This study intends to determine the extent of nuclear sperm injury in patients with varicocele and to investigate its relationship with apoptosis and oxidative stress (OS). Ejaculated sperm samples from 51 patients diagnosed with varicocele and 29 fertile men were examined. According to the guidelines, the patient's sperm samples were classified into varicocele with normal semen parameters (n = 11) and varicocele with abnormal semen parameters (n = 40). Sperm DNA fragmentation was assessed using terminal deoxynucleotidyl transferase dUTP nick end labeling assay. The proportion of both viable and dead spermatozoa with externalized phosphatidylserine (PS) was detected by the bivariate annexin V/6-CFDA staining method. Seminal malondialdehyde (MDA) amounts and antioxidant enzymes activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were measured spectrophotometrically. Sperm DNA fragmentation, viable spermatozoa with externalized PS, and MDA levels were significantly higher in studied subgroups of patients with varicocele, either with normal or with abnormal semen parameters than controls. The seminal antioxidant enzymes activities were significantly reduced in both subgroups of patients with varicocele compared to the controls. The percentage of spermatozoa with fragmented DNA was positively correlated to the MDA level as well as the proportion of viable spermatozoa with externalized PS. However, the decreased seminal antioxidant status was negatively correlated with the increased proportion of sperm DNA fragmentation and apoptotic spermatozoa. Impaired seminal antioxidant profile and increased seminal level of lipid peroxidation may be involved in the pathophysiological mechanisms of cell death-mediated DNA breaks in patients with varicocele.


Innervation of the posterior fossa dura of the cat.
This study was designed to identify the location of neurons giving rise to fibers innervating the posterior fossa dura in the cat using horseradish peroxidase (HRP, Sigma, Type VI). Investigations since the 19th century have implicated innervation by cranial nerves V, VII, IX, X and XII and the upper cervical nerves, C1-3. The meninges of the posterior fossa of 14 cats was exposed using one of three surgical approaches: (1) a suboccipital craniectomy and C1 laminectomy, (2) a parieto-occipital craniectomy with removal of the occipital lobe and bony tentorium exposing the meninges over the cerebellum, or (3) an anterior approach through the upper neck, exposing the dura of the ventral surface of the caudal brainstem. A unilateral, curvilinear incision was made in the dura and HRP was applied to the exposed dural edges. Following 48 hours the animals were sacrificed and fixed by perfusion. Cranial nerve ganglia of V, VII, IX, X, dorsal root ganglia (DRG) of C1-3, and superior cervical ganglia (SCG) were removed bilaterally, sectioned and processed with tetramethylbenzidine (TMB). HRP labeled cells were located bilaterally, always more ipsilaterally, in DRG of C1, C2, C3, and SCG with application of HRP to all three regions of the dura. Labeled cells were also located in trigeminal ganglia and superior ganglia of CN X, occasionally bilaterally, depending on the site of application. No HRP was ever identified in neurons of the geniculate ganglion, inferior ganglion of CN X or superior or inferior ganglia of CN IX. This information is valuable to an understanding of the innervation of intracranial structures and the problems of head pain.


Hair penalties: the negative influence of Afrocentric hair on ratings of Black women's dominance and professionalism.
Women are penalized if they do not behave in a stereotype-congruent manner (Heilman, 1983, 2001; Eagly and Carli, 2007). For example, because women are not expected to be agentic they incur an "agency penalty" for expressing anger, dominance or assertiveness (Rudman, 1998; Rudman and Glick, 1999, 2001; Eagly and Karau, 2002; Rudman and Fairchild, 2004; Brescoll and Uhlmann, 2008; Livingston et al., 2012). Yet, all women are not equally penalized (Livingston et al., 2012). We make a novel contribution by examining how both White and Black evaluators respond to displays of Black women's dominance, in this case, whether Black women choose to wear Afrocentric or Eurocentric hairstyles. We conducted three experimental studies to examine the influence of target hairstyle and participant race on ratings of the target's professionalism (Studies 1, 2, and 3) and dominance (Study 2). Study 1 was an online experimental study with 200 participants (112 females, 87 males, 1 missing gender; 160 Whites, 19 Blacks, 11 Latinos, 7 Asian Americans and 3 who identify as "other"; M age = 35.5, SD = 11.4). Study 2 was an online experimental study with 510 participants (276 women, 234 males; 256 Blacks, 254 Whites; M age = 41.25 years, SD = 12.21). Study 3 was an online experimental study with 291 participants (141 Blacks, 150 Whites, M age = 47.5 years, SD = 11.66). Black, as compared to White, evaluators gave higher agency penalties to Black employment candidates when they donned Afrocentric versus Eurocentric hair, rating them as more dominant and less professional. The present research illustrates the significance of considering both target and evaluator race when examining the influence of agency, and specifically dominance, on ratings of professionalism.


Total Synthesis of the Aminopropyl Functionalized Ganglioside GM(1).
GM1 is a common ganglioside pentasaccharide present on mammalian cell surface. It has been shown to play important roles in cellular communications and initiation of β-amyloid aggregation. In order to synthesize GM1, an efficient synthetic route was developed via a [3+2] strategy. The GM3 trisaccharide acceptor bearing an azido propyl group at the reducing end was prepared using the traditional acetamide protected sialyl thioglycosyl donor, which gave better stereoselectivity than sialyl donors protected with trichloroacetamide or oxazolidinone. The glycosylation of the axial 4-hydroxyl group of GM3 by the disaccharide donor was found to be highly dependent on donor protective groups. Donor bearing the more rigid benzylidene group gave low glycosylation yield. Replacing the benzylidene with acetates led to productive coupling and formation of the fully protected GM1 pentasaccharide. Deprotection of the pentasaccharide produced GM1 functionalized with the amino propyl side chain, which will be a valuable probe for biological studies.


Fibrolipomatous Hamartoma of the Median Nerve: An Outcome of Surgical Management in Six Consecutive Cases.
Lipoma is a nonneurogenic benign tumor. Neurolipoma and fibrolipomatous hamartoma are variants of this universal tumor. All these variants are grouped under lipomatosis of the nerve. Majority of these tumors are asymptomatic, which can be observed. Symptomatic patients require surgery, which is not standardized. As there are insufficient number of cases, no randomized controlled studies have been performed in the treatment of fibrolipomatous hamartoma. The aim of our study was to determine the pattern of presentation of fibrolipomatous hamartoma, surgical management offered, and the outcome in the form of recovery and complications. This retrospective descriptive study includes six patients diagnosed with fibrolipomatous hamartoma over a period of 12 years. Patient details were collected from the medical records. Patients diagnosed of fibrolipomatous hamartoma in the hand were included. Patients with other soft-tissue tumors were excluded from the study. Out of six patients, four required excision of nerve followed by reconstruction using sural nerve graft and two underwent microsurgical dissection of neural element. Patients were instructed to take care of the operated hand during the recovery phase. Institutional physiotherapy protocol was started during the 3rd postoperative week. Follow-up period was between 1 and 3 years. All the six patients were free from symptoms postoperatively. Minimal complications were noted in two patients, which were managed conservatively. Surgical excision of fibrolipomatous hamartoma of median nerve below elbow, with nerve dissection or with nerve reconstruction using sural nerve graft, followed by proper postoperative care and physiotherapy has proven beneficial for the patients in our study.


High- versus low-dose ACE inhibition in chronic heart failure: a double-blind, placebo-controlled study of imidapril.
To determine dose-related clinical and neurohumoral effects of angiotensin-converting enzyme (ACE) inhibitors in patients with chronic heart failure (CHF), we conducted a double-blind, placebo-controlled, randomized study of three doses (2.5 mg, 5 mg and 10 mg) of the long-acting ACE inhibitor imidapril. The ACE inhibitors have become a cornerstone in the treatment of CHF, but whether high doses are more effective than low doses has not been fully elucidated, nor have the mechanisms involved in such a dose-related effect. In a parallel group comparison, the effects of three doses of imidapril were examined. We studied 244 patients with mild to moderate CHF (New York Heart Association class II-III: +/-80%/20%), who were stable on digoxin and diuretics. Patients were treated for 12 weeks, and the main end points were exercise capacity and plasma neurohormones. At baseline, the four treatment groups were well-matched for demographic variables. Of the 244 patients, 25 dropped out: 3 patients died, and 9 developed progressive CHF (3/182 patients on imidapril vs. 6/62 patients on placebo, p < 0.05). Exercise time increased 45 s in the 10-mg group (p = 0.02 vs. placebo), but it did not significantly change in the 5-mg (+16 s), and 2.5-mg (+11 s) imidapril group, compared to placebo (+3 s). Physical working capacity also increased in a dose-related manner. Plasma brain and atrial natriuretic peptide decreased (p < 0.05 for linear trend), while (nor)epinephrine, aldosterone and endothelin were not significantly affected. Renin increased in a dose-related manner, but plasma ACE activity was suppressed similarly (+/-60%) on all three doses. Already within 3 months after treatment initiation, high-dose ACE inhibition (with imidapril) is superior to low-dose. This is reflected by a more pronounced effect on exercise capacity and some of the neurohormones, but it does not appear to be related to the extent of suppression of plasma ACE.


Reexamining the role of choline transporter-like (Ctlp) proteins in choline transport.
In Saccharomyces cerevisiae, choline enters the cell via a single high-affinity transporter, Hnmlp. hnm1delta cells lacking HNM1 gene are viable. However, they are unable to transport choline suggesting that no additional active choline transporters are present in this organism. A complementation study of a choline auxotrophic mutant, ctrl-ise (hnm1-ise), using a cDNA library from Torpedo marmorata electric lobe identified a membrane protein named Torpedo marmorata choline transporter-like, tCtl1p. tCtllp was proposed to mediate a high-affinity choline transport (O'Regan et al., 1999, Proc. Natl. Acad. Sci.). Homologs of tCtl1p have been identified in other organisms, including yeast (Pns1p, YOR161c) and are postulated to function as choline transporters. Here we provide several lines of evidence indicating that Ctlp proteins are not involved in choline transport. Loss of PNS1 has no effect on choline transport and overexpression of either PNS1 or tCTL1 does not restore choline uptake activity of choline transport-defective mutants. The data presented here call into question the role of proteins of the CTL family in choline transport and suggest that the mechanism by which tCTL1 complements hnm1-ise mutant is independent of its ability to transport choline.


Development and validation of an LC-MS/MS method for the determination of SB-505124 in rat plasma: Application to pharmacokinetic study.
A sensitive, selective and rapid liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-MS/MS) method has been developed for the quantification of the novel transforming growth factor-β (TGF-β) inhibitor SB-505124 in rat plasma and then validated. Plasma samples were prepared by simple protein precipitation. Separation was performed on a Diamonsil ODS chromatography column using a mobile phase of acetonitrile and 0.1% (v/v) aqueous formic acid. SB-505124 and the internal standard doxorubicin were detected in the positive ion mode using multiple reaction monitoring of the transitions at m/z 336.2→320.1 and 544.2→397.2, respectively. Calibration curve was linear (r>0.9996) over a concentration range of 10-5000 ng/mL with the lower quantification limit of 10 ng/mL. Both intra- and inter-day precision were within 6.5% and trueness were not more than 3.1%. Extraction recovery and matrix effect were within acceptable limits. Stability tests showed that SB-505124 and the IS remained stable throughout the analytical procedure. The validated LC-MS/MS method was then used to analyze the pharmacokinetics of SB-505124 administered to rats intravenously (8 mg/kg) or orally (10 mg/kg). Oral bioavailability of SB-505124 was calculated as 76.4%, indicating the potential of SB-505124 as an orally administered drug.


The efficacy of progesterone 1 mg kg-1 every 12 hours over 5 days in moderate-to-severe traumatic brain injury: A meta-analysis of randomized controlled trials.
The objective of this meta-analysis was to assess the efficacy of progesterone (PG) at 1.0 mg kg-1 once every 12 h for 5 consecutive days in patients with moderate-to-severe (Glasgow Coma Score [GCS] 3-12) traumatic brain injury (TBI). The Cochrane Library, OvidSP, Web of Science, PubMed, CNKI, WFSD, and VIP databases were systematically searched from inception to May 1, 2020. The quality of included studies was evaluated using the bias risk assessment tool from the Cochrane systematic evaluator manual 5.1.0. A pooled analysis of relevant data was conducted using RevMan 5.3 software. The primary outcome was good functional outcome (GFO), and the secondary outcome was mortality. Subgroup analysis was performed to explore the impact of time, administration route, type of injury, and GCS on outcome measures. A total of 7 randomized controlled trials involving 504 participants were included in this meta-analysis. The findings indicated a statistically significant difference in terms of GFO (RR, 1.48; 95 % confidence interval [CI], 1.25-1.76; P < 0.00001) and mortality (RR, 0.66; 95 % CI, 0.44-0.84; P = 0.002) between the PG and control groups. Subgroup analyses demonstrated that administration route was an important influencing factor for improving GFO in the PG group, and administration route and follow-up time were important for reducing mortality in the PG group. We conclude that PG, at a dose of 1.0 mg kg-1 via intramuscular injection every 12 h for 5 consecutive days, could significantly improve GFO (1 month, 3 months, 6 months, and 12 months) and reduce the medium-term (3-month and 6-month) mortality rate. Larger studies are needed to support our findings.


New ultrastructural evidence for a protein transport system in endothelial cells of gerbil brains.
Pathways for transfer of horseradish peroxidase (HRP) across the cerebral microvasculature were studied in Mongolian gerbils after inducing either unilateral carotid-artery ligation or intracarotid air embolism. Electron microscopy on samples from both ipsilateral and contralateral brain hemispheres showed the reaction product (HRP) in the endothelial cytoplasm of capillaries and arteriols in all animals; in the basal lamina, HRP was seen only after long (3--4 h) ischemic periods. HRP was seen within both spherical and tubular structures, with the latter appearing in obliquely or longitudinally sectioned blood vessels. These endothelial channel-like structures are part of a tubulovesicular network which seemingly exists in the normal endothelium of the gerbil brain. The tubules serve as pathways for proteins into the brain; protein transfer becomes enhanced following circulatory injuries such as ischemia.


Solubilization and resolution of thromboxane synthesizing system from microsomes of bovine blood platelets.
The thromboxane synthetase system of the microsomes of bovine blood platelets was solubilized by the treatment with Triton X-100. The solubilized preparation was separated into two enzyme fractions by DEAE-cellulose chromatography. One catalyzed the formation of prostaglandin H2 from arachidonic acid in the presence of heme and tryptophan. The other fraction converted prostaglandin H2 to thromboxane B2 and 12L-hydroxy-5,8,10-heptadecatrienoic acid. However, incubation of the latter fraction with prostaglandin H2 at lower temperature produced an unstable compound with platelet-aggregating activity, which was presumably thromboxane A2 and which decomposed readily to thromboxane B2 and 12L-hydroxy-5,8,10-heptadecatrienoic acid.


Factors associated with wrong blood in tube errors: An international case series - The BEST collaborative study.
A wrong blood in tube (WBIT) error signifies a blood sample that does not match the patient identified on the sample label. WBIT errors can result in ABO mistransfusions. In this international, multicenter, descriptive study, healthcare facilities provided detailed information on WBIT errors occurring from 1/1/2019 to 12/31/2020. Factors contributing to WBIT errors were classified as protocol violations, knowledge gaps, and slips/lapses. 331 WBIT errors were compiled from 36 centers in 11 countries. WBIT errors were most frequently detected through pretransfusion sample testing (191, 58%), with 38 (20%) detected by a second ("check") sample. WBIT errors were divided almost evenly between intended patient drawn/wrong label applied (166, 50%) and wrong patient drawn/intended label applied (158, 48%). Information on contributing factors was available for 260 WBIT errors; most involved a combination of protocol violations and slips/lapses (139, 53%). The most frequent contributing factor was another patient's sample labels or tubes being available during phlebotomy (61%). Protocol violations were more likely to result in wrong patient being drawn (p = .0007). In 43 WBIT errors, electronic positive patient identification (ePPID) was not used when available or was used incorrectly. Protocol violations and slips/lapses frequently contribute to WBIT errors. Sample collection processes should be designed to minimize error opportunities; staff should be educated on why protocol compliance is critical for patient safety. Using ePPID does not eliminate all WBIT errors. Institutions using ePPID may elect to require check sample verification as an added safety measure.


Treatment of the intestinal tuberculosis.
Twenty-seven cases of patients with intestinal tuberculosis were reported. Seventeen cases showed good response to antituberculosis chemotherapy. The numbers and size of ulceration, pseudopolyp and deformity of intestinal tract were checked by endoscopic and X-ray studies on comparative evaluation between prechemotherapeutic and postchemotherapeutic stage. In these cases, subjective symptoms were disappeared parallel with the marked improvement on X-ray and endoscopic findings. These data suggest that with current well planned antituberculosis chemotherapy program, tuberculous colitis is controlled fairly while tuberculous enteritis in controlled inadequately. Therefore, surgical procedures are commly indicated to the treatment of tuberculous enteritis.


Characterization of high yield exopolysaccharide produced by Phyllobacterium sp. 921F exhibiting moisture preserving properties.
A new strain bacteria was isolated and named as Phyllobacterium sp. 921F, due to its high production capacity of exopolysaccharide (EPS). Characterization of physico-chemical properties of the EPS and optimization for high production were conducted to aim at industrial applications. The optimum pH and temperature were 7.0 and 30°C, respectively. The following scale-up fermentation was carried out in 30L bioreactor and amounts of EPS (21.9g/L) were produced. The EPS with a molecular mass of 1082kDa was composed of glucose, galactose, and pyruvate. The EPS solution behaved as Newtonian at low concentrations (≤0.3%) and as shear thinning at higher concentration (e.g, 1%). The moisture retention ability of the EPS was found to be superior to hyaluronic acid. Results suggest that Phyllobacterium sp. 921F is a good candidate for large-scale production of the EPS which might be utilized in food and cosmetics industries.


Evaluation of Aneurysm Neck Angle Change After Endovascular Aneurysm Repair Clinical Investigations.
To evaluate the aneurysm neck angle changes and post-endovascular aneurysm repair (EVAR) complications. We retrospectively analyzed 72 cases of elective EVAR for abdominal aortic aneurysm among 109 consecutive cases from December 2005 to April 2014. Patients were divided into angulated and non-angulated groups. The angulated group was defined as neck angulation ≥60°. Neck angle was evaluated pre- and post-EVAR during short- (within 1 month), mid- (3-6 months), and long-term (>1 year) follow-up. Aneurysm sac diameter change, aneurysm neck morphology other than angulation, endoleaks, and other post-procedural complications were also documented. A total of 34 patients were enrolled in the angulated group. There were no statistical differences in age, sex, follow-up duration, and aneurysm neck profile between the two groups (p > 0.05). Both groups showed statistically significant and consistent decreases in angulation during the follow-up period (p < 0.01). The angulated group revealed 22.45 % more straightening than the non-angulated group. Recoil of the Endurant device occurred in the angulated group. No statistically significant intergroup differences were observed in any endoleaks, complications, or re-intervention rates (p > 0.05). Pre-EVAR angle was the only predictor for post-procedural angle change (p < 0.001). EVAR is applicable for patients with highly angulated aneurysm neck and provides consistent neck straightening over long-term follow-up. Recoil was evident in the angulated group using the Endurant device.


Itaconate derivative 4-OI inhibits PRRSV proliferation and associated inflammatory response.
Itaconate, a metabolite of the tricarboxylic acid (TCA) cycle produced by immunoresponsive gene 1 (IRG1) via catalyzation of cis-aconitate, plays important roles in metabolism and immunity. Porcine reproductive and respiratory syndrome virus (PRRSV) is an Arterivirus that has devastated the swine industry worldwide for over 30 years. Here, we found that 4-octyl itaconate (4-OI), a cell-permeable itaconate derivative, dose-dependently inhibited PRRSV proliferation by interfering with viral attachment, replication, and release. Furthermore, 4-OI suppressed the PRRSV-induced inflammatory response by enhancing nuclear factor erythroid 2-related factor 2 (Nrf2) signaling. Interestingly, PRRSV infection caused a reduction in itaconate abundance and simultaneously led to an accumulation of cis-aconitate, the upstream metabolite of itaconate, and both of these effects were accomplished by downregulating IRG1 expression. Taken together, these results demonstrate that 4-OI not only inhibits PRRSV replication but also suppresses PRRSV-induced inflammatory responses, indicating that 4-OI is a promising drug candidate for combating PRRSV.


Risk Factors for In-Hospital Cardiac Arrest in Patients With ST-Segment Elevation Myocardial Infarction.
In-hospital cardiac arrest (IHCA) is one of the most deleterious complications of ST-segment elevation myocardial infarction (STEMI). We systematically analyzed the clinical characteristics of STEMI patients with IHCA, as well as predictors and treatments associated with risk of IHCA, using a nationwide database. In the CCC-ACS (Improving Care for Cardiovascular Disease in China-Acute Coronary Syndrome) project (2014-2019), we stratified patients presenting with STEMI within 24 hours after symptom onset according to IHCA or no IHCA during the index hospitalization. We analyzed patients' clinical characteristics, mortality, and independent correlates of IHCA. Of 40,670 STEMI patients, 2.2% (95% CI: 2.1%-2.4%) experienced IHCA. Among IHCA patients, the in-hospital mortality was 53.0% (95% CI: 49.7%-56.3%). IHCA represents 55.0% (95% CI: 51.6%-58.4%) of inpatient deaths. Age ≥75 years, female, nonsmoker, prior diabetes mellitus, prior renal failure, out-of-hospital cardiac arrest, heart rate >100 beats/min, systolic blood pressure <90 mm Hg, and Killip IV were identified as predictors of IHCA. IHCA patients were less likely to receive β-blockers and ticagrelor during the first 24 hours after first medical contact and were less likely to undergo primary percutaneous coronary intervention. After adjustment, primary percutaneous coronary intervention (adjusted HR: 0.82; 95% CI: 0.71-0.95), β-blockers (adjusted HR: 0.63; 95% CI: 0.47-0.86), and ticagrelor (adjusted HR: 0.57; 95% CI: 0.42-0.76) were associated with a reduced risk of IHCA. IHCA is rare in STEMI but is associated with high mortality. Multiple modifiable and unmodifiable factors are associated with its occurrence, suggesting that early intervention and rational drug treatment may improve its prognosis. (CCC Project- Acute Coronary Syndrome; NCT02306616).


Ultrastructural changes of polypropylene loops in a chronically inflamed eye.
We have documented diffuse, superficial, transverse fissures of blue polypropylene IOL loops that had been exposed to a prolonged inflammatory reaction. The cause and clinical significance of this process has not yet been determined. The polymethylmethacrylate optic showed no similar changes.


Lipid-lowering drugs and serum liver enzymes: the effects of body weight and baseline enzyme levels.
The most important side effects of fibrate and 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor (statin) treatment are hepatic toxicity and myopathy. Obese individuals may have higher levels of serum transaminases than their lean counterparts. The main purpose of this study was to examine the effects of statins and fibrates on liver enzymes in obese patients and to compare them with their effects on patients with various body mass indexes (BMI). Two hundred and sixty-three hyperlipidemic patients of both sexes aged 31-74 years were studied for 24 weeks. One hundred and three patients received fluvastatin (40 mg/day), 62 atorvastatin (10-20 mg/day), 45 micronized fenofibrate (200 mg/day), 44 ciprofibrate (100 mg/day) and nine patients received gemfibrozil (900 mg/day). Laboratory determinations were performed at baseline, after 8 weeks of treatment and at the end of the follow-up period. At baseline, obese patients tended to exhibit elevated liver enzymes more frequently than their lean counterparts (12 of 105 vs. 5 of 67). At the end of the study period, 11 obese, seven overweight and six lean subjects exhibited elevated liver enzymes. Twelve patients who experienced a moderate elevation of serum liver enzymes at baseline had their liver enzyme profile normalized at the end of the study. Furthermore, in 12 patients who had normal serum liver enzyme levels at baseline, abnormal levels of at least one enzyme were observed after 24 weeks of treatment. Fibrates and statins are safe drugs for the treatment of hyperlipidemia in obese patients as well as in those with moderately increased liver enzymes.


A less invasive approach to completely repair the aortic arch.
Surgical replacement of the aortic arch is an established procedure that requires cardiopulmonary bypass and deep hypothermic circulatory arrest. However, this approach is associated with major perioperative risks. The significant risks associated with conventional open repair of the thoracic aorta are the main argument for less invasive strategies. We present a less invasive surgical and endovascular approach that allows total exclusion of the aortic arch without the need for extracorporeal circulation, deep hypothermia, and circulatory arrest. All procedures were successful, and the patients recovered without neurologic, cardiac, or bleeding complications. Arteriography confirmed proper position of the stent graft and complete exclusion of the lesion at the end of the procedure. One patient had an endoleak type I and underwent successful additional retrograde stent-graft placement over the proximal landing zone 3 weeks after the initial procedure. Clinical follow-up (between 8 and 18 months) was fully uncomplicated in all patients. Computed tomographic scan at 6 months demonstrated complete exclusion of the arch lesion in all cases and did not reveal any endoleaks. Assuming that technical refinements may improve all steps of the endovascular intervention, this combined approach may turn out to be the preferred therapeutic modality to repair aortic arch lesions in patients with multiple comorbidities who otherwise would not be candidates for a conventional operative repair. However, long-term observation is necessary to confirm the stability of this type of repair.


Ethanol and Isopropyl Alcohol Exposure Increases Biofilm Formation in Staphylococcus aureus and Staphylococcus epidermidis.
Alcohols, including ethanol and isopropyl alcohol, are used in clinical practice for disinfection and infection prevention. Recent studies, however, demonstrate that alcohols may enhance biofilm production in Staphylococci. We quantified biofilm formation in the presence of ethanol and isopropyl alcohol in six different, well-characterized strains of Staphylococcus epidermidis and Staphylococcus aureus. After 24 h of biofilm development, each strain was exposed to normal saline (NS), ethanol, or isopropyl alcohol (40%, 60%, 80% and 95%) for additional 24 h incubation. Adherent biofilms were stained and optical density was determined. Viability of strains was also determined after alcohol exposure. Ethanol increased biofilm formation in all six strains compared to normal saline (p < 0.05). There was increased biofilm formation with increasing ethanol concentration. Isopropyl alcohol also increased biofilm formation with increasing alcohol concentration in all six strains (p < 0.01 vs NS). The slime-negative, chemical mutant strain of S. epidermidis increased biofilm formation after exposure to both alcohols, likely reverting back its primary phenotype through modulation of the intercellular adhesin repressor. All strains demonstrated viability after exposure to each alcohol concentration, though viability was decreased. Ethanol and isopropyl alcohol exposure increases biofilm formation of S. aureus and S. epidermidis at concentrations used in clinical settings. Ethanol and isopropyl alcohol did not eradicate viable Staphylococci from formed biofilm.


Trends in Routine Opioid Dispensing After Common Pediatric Surgeries in the United States: 2014-2019.
Using Joinpoint regression, our study revealed substantial decreases in postoperative opioid dispensing after outpatient pediatric surgeries beginning in 2017.


Quantitative cancer risk assessments for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).
State-of-the-art quantitative risk assessment techniques, including consideration of new time-to-response data, have been applied to chronic animal bioassay data on the dietary intake of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). The non-linear shapes of the dose-response relationships for the hepatocellular carcinogenic responses have been estimated, and a review of the quantitative impacts of several of the choices involved in the quantitative risk assessment considers, particularly, the definition of the carcinogenic responses of concern, the experimental data set, the pathology evaluation, a biologically effective dose scale versus the administered dose, methods of making the fitted model responsive to the data at the lower experimental doses, consistency in dose-response shapes for different data sets, fitted model values versus bounds, the utilization of time-to-response information incorporating the lateness of the carcinogenic responses, and the method of characterizing the maximum acceptable dose. The estimated virtually safe dose for an increase of 0.000001 (one in a million) in the probability of hepatocellular neoplastic nodule and/or carcinoma in a female rat is approximately 0.1 ng/kg body weight/day in the diet. The estimated mean free dose, corresponding to a reduction in the expected amount of time without hepatocellular neoplastic nodule and/or carcinoma proportional to 1 wk in 70 yr, is in the range of 1-5 ng/kg body weight/day in the diet of a female rat. No species-to-species extrapolations nor human exposure assessments have been made. However, these estimated risks correspond to dietary intakes that are at least 150 times greater than the 0.0006365 ng/kg body weight/day intake described by the Centers for Disease Control as a reasonable level to begin consideration of action to limit human exposure.


National Prevention through Design (PtD) Initiative.
The most effective means of preventing and controlling occupational injuries, illness, and fatalities is to "design out" hazards and hazardous exposures from the workplace. There is a long history of designing for safety for the general public and to a lesser degree for workers. We now have the experience and insight from thoughtful, previous efforts to call for a comprehensive national strategy to implement a Prevention through Design (PtD) Initiative. This paper describes that initiative in terms of four overarching areas where action can be directed: practice, policy, research, and education. To obtain stakeholder input for issues in these four areas and to focus implementation efforts, eight sector divisions of the economy will be addressed. A seven year strategy is envisioned.


Factors influencing student nurses' readiness to adopt artificial intelligence (AI) in their studies and their perceived barriers to accessing AI technology: A cross-sectional study.
With the projected significant increase in the use of AI in nursing education, it becomes vital for nurse faculty to adequately equip student nurses with the necessary competences to effectively utilize AI in their studies. Ensuring that student nurses are prepared and ready to embrace AI technology is imperative for their successful integration into the healthcare workforce. This study aimed to examine student nurses' readiness to embrace AI technology, explore associated factors, and identify perceived barriers to accessing AI technology. Cross sectional study. One public-owned nursing school in the Philippines. Three hundred twenty-three student nurses. Data were collected using structured questionnaires. Descriptive statistics and multivariable analysis were performed to analyze the data. The results revealed that student nurses demonstrated moderate readiness to embrace AI in their studies (M = 2.906, SD = 0.692) and perceived moderate barriers to accessing AI technology (M = 2.336, SD = 0.719). Factors associated with students' readiness to embrace AI included self-rated technological proficiency (β = 0.170, p = 0.014), understanding of AI-powered technologies (β = 0.260, p < 0.001), and perceived AI use in nursing practice (β = 0.153, p = 0.022). The study also identified potential barriers to accessing AI technology, such as lack of computer skills to navigate AI, lack of AI knowledge and awareness, and time constraints. The findings of this study provided valuable insights into the factors influencing student nurses' attitudes towards AI and shed light on their perceived barriers to accessing AI technology. By enhancing technological proficiency, increasing AI understanding, and providing practical experiences, nurse faculty can better prepare future nurses to effectively navigate the AI-driven healthcare environment and contribute to improved patient care outcomes.


Maximal oxygen uptake, anthropometry and physical activity in a randomly selected sample of 8 and 13 year old children in Sweden.
Maximal oxygen uptake was assessed in 101 randomly selected 8 and 13 year old children. In both age groups a significantly higher aerobic capacity was found in boys than in girls, both in absolute terms and when maximal oxygen uptake was related to body weight, lean body mass and lean leg volume. Among girls, maximal oxygen uptake per kg body weight was lower in the older than in the younger (p less than 0.05). Estimation of spontaneous physical activity, by means of a questionnaire and the actometry method, indicated that physical activity was greater in children with a high than in those with a low aerobic capacity.


Pharmacogenetic Risk Stratification in Angiotensin-Converting Enzyme Inhibitor-Treated Patients with Congestive Heart Failure: A Retrospective Cohort Study.
Evidence for pharmacogenetic risk stratification of angiotensin-converting enzyme inhibitor (ACEI) treatment is limited. Therefore, in a cohort of ACEI-treated patients with congestive heart failure (CHF), we investigated the predictive value of two pharmacogenetic scores that previously were found to predict ACEI efficacy in patients with ischemic heart disease and hypertension, respectively. Score A combined single nucleotide polymorphisms (SNPs) of the angiotensin II receptor type 1 gene (rs275651 and rs5182) and the bradykinin receptor B1 gene (rs12050217). Score B combined SNPs of the angiotensin-converting enzyme gene (rs4343) and ABO blood group genes (rs495828 and rs8176746). Danish patients with CHF enrolled in the previously reported Echocardiography and Heart Outcome Study were included. Subjects were genotyped and categorized according to pharmacogenetic scores A and B of ≤1, 2 and ≥3 each, and followed for up to 10 years. Difference in cumulative incidences of cardiovascular death and all-cause death were assessed by the cumulative incidence estimator. Survival was modeled by Cox proportional hazard analyses. We included 667 patients, of whom 80% were treated with ACEIs. Differences in cumulative incidences of cardiovascular death (P = 0.346 and P = 0.486) and all-cause death (P = 0.515 and P = 0.486) were not significant for score A and B, respectively. There was no difference in risk of cardiovascular death or all-cause death between subjects with score A ≤1 vs. 2 (HR 1.03 [95% CI 0.79-1.34] and HR 1.11 [95% CI 0.88-1.42]), score A ≤1 vs. ≥3 (HR 0.80 [95% CI 0.59-1.08] and HR 0.91 [95% CI 0.70-1.20]), score B ≤1 vs. 2 (HR 1.02 [95% CI 0.78-1.32] and HR 0.98 [95% CI 0.77-1.24]), and score B ≤1 vs. ≥3 (HR 1.03 [95% CI 0.75-1.41] and HR 1.05 [95% CI 0.79-1.40]), respectively. We found no association between either of the analyzed pharmacogenetic scores and fatal outcomes in ACEI-treated patients with CHF.


Exploring undergraduate medical students' perception of learning procedural skills and its outcomes in clinical settings.
Learning procedural skills is one of the essential aspects of undergraduate medical education. However, learning procedural skills in clinical settings is less widely considered. This study aimed to explore the Iranian undergraduate medical students' perception of learning procedural skills and its outcomes in three universities of medical sciences in Iran. A descriptive exploratory qualitative methodology with an in-depth unstructured, face-to-face interview, and content analysis was used in this study. Sixteen students in clinical phases of general medical education programs from educational hospitals were selected using purposive sampling. According to the preferences of the participants, the interviews were conducted in medical schools or in hospitals. The students participating in this study included 7 females and 9 males (totally 16 people) with a mean age of 23.7 years old with a range of 21-27 years. The three main themes of this study were "the gap of transferring formal teaching from skill lab to clinical placement", "learning self-leading procedural skills in clinical settings", and "students' dissatisfaction with patients' vulnerability" with 8 subthemes which were extracted and explained based on the students' perception. Unsupervised and self-learning by medical students and weakness in controlling the learning process have undesirable results for patients and students.


Prevalence of hypertension and associated risks in hospitalized patients with COVID-19: a meta-analysis of meta-analyses with 1468 studies and 1,281,510 patients.
Since the COVID-19 outbreak, preliminary research has shown that some risk-associated conditions increase death and severe complications of the disease, hypertension being one of them. Thus, numerous meta-analyses have been conducted to explore this issue. Therefore, this umbrella review aims to perform a meta-analysis of the meta-analyses to estimate the prevalence and associated risks of hypertension in patients with COVID-19. PubMed, Scopus, Web of Knowledge, Embase, and Cochrane databases were searched for the published meta-analyses up to January 1, 2022. Google Scholar, citation check, reference check, and Grey literature were also manually searched. A random-effect model approach was used for analysis. The overall death rate was estimated at 12%. Hypertension was present in 25% of the patients as a comorbid disease. The overall RR for death, disease severity, and the possibility of ICU admission were estimated at 1.79 [1.68-1.89 with 95% CI], 1.74 [1.66-1.83 with 95% CI], and 1.91 [1.48-2.34 with 95% CI], respectively. The meta-regression results showed that being "male" significantly increases the risk of disease severity and ICU admission. The results indicated that hypertension is a common comorbid disease in hospitalized patients with COVID-19, which significantly increases mortality risk, the severity of the disease, and the probability of ICU admission. This study has been registered in PROSPERO (CRD42021231844).


Management of corrosive esophageal burns in 149 cases.
We sought to present our experience in the management of esophageal burns. From April 1976 through October 2003, 149 patients with corrosive esophageal burns were included in this study. Treatment modalities consisted of modified intraluminal stenting in 28, colon interposition in 71, gastric transposition in 25, repair of cervical stricture with platysma myocutaneous flap in 17, and miscellaneous operations in 12 patients. Eleven of these patients underwent the above procedures twice at our institute. The remaining 7 patients were treated with conservative therapy. Twenty-three patients recovered from intraluminal stenting, and 5 experienced stricture after stent removal. One of the 5 patients with failed stents responded to bougienage, and the remaining 4 patients required esophageal reconstruction later. Of the 71 colon interpositions, 5 patients died postoperatively, and complications consisted of proximal anastomotic fistula in 17, anastomotic stenosis in 6, and abdominal incision dehiscence in 2 patients. Postoperative complications in the 25 patients with gastric transpositions comprised anastomotic stricture in 2 patients and empyema in 1 patient. There was a cervical leak in 1 of the 17 patients undergoing the repair of cervical esophageal or anastomotic stricture with a platysma myocutaneous flap. One of the patients in the group undergoing 12 miscellaneous procedures died 8 months after surgical intervention. All the survivors currently eat regular diets. Intraluminal stenting can prevent the formation of caustic esophageal stricture. The location of the cicatricial esophagus dictates whether to perform concomitant esophagectomy during esophageal reconstruction. Platysma myocutaneous flap repair is an excellent method for the treatment of severe cervical esophageal or anastomotic stricture.


Aligning the 3Rs with new paradigms in the safety assessment of chemicals.
There are currently several factors driving a move away from the reliance on in vivo toxicity testing for the purposes of chemical safety assessment. Progress has started to be made in the development and validation of non-animal methods. However, recent advances in the biosciences provide exciting opportunities to accelerate this process and to ensure that the alternative paradigms for hazard identification and risk assessment deliver lasting 3Rs benefits, whilst improving the quality and relevance of safety assessment. The NC3Rs, a UK-based scientific organisation which supports the development and application of novel 3Rs techniques and approaches, held a workshop recently which brought together over 20 international experts in the field of chemical safety assessment. The aim of this workshop was to review the current scientific, technical and regulatory landscapes, and to identify key opportunities towards reaching these goals. Here, we consider areas where further strategic investment will need to be focused if significant impact on 3Rs is to be matched with improved safety science, and why the timing is right for the field to work together towards an environment where we no longer rely on whole animal data for the accurate safety assessment of chemicals.


The Duchenne muscular dystrophy gene product is localized in sarcolemma of human skeletal muscle.
Duchenne muscular dystrophy (DMD) and its milder form, Becker muscular dystrophy (BMD), are allelic X-linked muscle disorders in man. The gene responsible for the disease has been cloned from knowledge of its map location at band Xp21 on the short arm of the X chromosome. The product of the DMD gene, a protein of relative molecular mass 400,000 (Mr 400K) recently named dystrophin, has been reported to co-purify with triads of mouse and rabbit skeletal muscle when assayed using polyclonal antibodies raised against fusion proteins encoded by regions of mouse DMD complementary DNA. Here we show that antibodies directed against synthetic peptides and fusion proteins derived from the N-terminal region of human DMD cDNA strongly react with an antigen present in skeletal muscle sarcolemma on cryostat sections of normal human muscle biopsies. This immunoreactivity is reduced or absent in muscle fibres from DMD patients but appears normal in muscle fibres from patients with other myopathic diseases. The same antibodies specifically react with a 400K protein in sodium dodecyl sulphate (SDS) extracts of normal human muscle subjected to Western blot analysis. We conclude that the product of the DMD gene is associated with the sarcolemma rather than with the triads and speculate that it strengthens the sarcolemma by anchoring elements of the internal cytoskeleton to the surface membrane.


Recurrent Clostridium difficile infection: From colonization to cure.
Clostridium difficile infection (CDI) is increasingly prevalent, dangerous and challenging to prevent and manage. Despite intense national and international attention the incidence of primary and of recurrent CDI (PCDI and RCDI, respectively) have risen rapidly throughout the past decade. Of major concern is the increase in cases of RCDI resulting in substantial morbidity, morality and economic burden. RCDI management remains challenging as there is no uniformly effective therapy, no firm consensus on optimal treatment, and reliable data regarding RCDI-specific treatment options is scant. Novel therapeutic strategies are critically needed to rapidly, accurately, and effectively identify and treat patients with, or at-risk for, RCDI. In this review we consider the factors implicated in the epidemiology, pathogenesis and clinical presentation of RCDI, evaluate current management options for RCDI and explore novel and emerging therapies.


Visual loss associated with choroidal nevi.
Twenty-two of 206 patients (11%) with choroidal lesions diagnosed clinically as nevi of the posterior fundus had decreased visual acuity attributable to the lesion. The decreased visual acuity was secondary to serous foveal detachment (50%), presumed photoreceptor degeneration without serous detachment (42%), and subretinal choroidal neovascularization overlying the nevus (8%). Visual loss associated with a small melanotic choroidal tumor does not necessarily imply that the tumor is a choroidal malignant melanoma as 88% of the small melanotic choroidal tumors causing visual impairment did not grow during an average follow-up period of 25 months, and the great majority of these tumors were probably true spindle cell nevi. However, three tumors (12%), initially categorized as "suspicious nevi," enlarged during follow-up and were recategorized as malignant melanomas.


Isolation and characterization of cardiac sarcolemma.
A procedure was developed for the isolation of cardiac sarcolemmal vesicles. These vesicles are enriched about ten-fold (with respect to the tissue homogenate) in K+-stimulated p-nitrophenylphosphatase, (Na+ + K+)-ATPase, 5'-nucleotidase activities and sialic acid content, all of which are believed to be components of the sarcolemma. The sarcolemma of tissue culture cardiac cells were radioiodinated and the distribution of this radioiodine paralleled the distribution of the other membrane markers above. There was very little contamination of the sarcolemmal fraction by sarcoplasmic reticulum (as judged by Ca2+-ATPase and glucose-6-phosphatase activities) or inner mitochondrial membranes (as judged by succinate dehydrogenase activity). There may, however, be some contamination by outer mitochondrial membranes (as judged by monoamine oxidase and rotenone-insensitive NADH cytochrome c reductase activities) which have rarely been monitored in cardiac sarcolemmal preparations. The purity of this preparation is good when compared with other cardiac sarcolemmal preparations. This preparation should be very useful in studying the roles of the cardiac sarcolemma (e.g. in excitation contraction coupling and Ca2+ binding).


[Transcervical endometrial resection. A new surgical method for the management of metrorrhagia].
The authors introduce to transcervical endometrial resection offered as an alternative operation for women with metrorrhagia. Their experience regarding to the introduction and results of the surgery of the first 10 patients are reported. There were no intraoperative complications. The benefit of the new technique regarding to the shorter hospital stay and recovery time is obvious.


Geometrical factors influencing muscle force development. II. Radial forces.
If the subfragment-2 (S2) portion of the myosin cross-bridge to actin does not lie parallel to the myofilament axes then when a muscle fiber contracts, there will be a radial component to the cross-bridge force. When the subfragment-1 (S1) portion of the cross-bridge attaches to actin with its long axis projecting through the filament axis, the magnitude of the radial force depends upon the azimuthal location of the actin site, but when the attachment of the S1 to actin is slewed, as in the reconstruction of Moore et al. (J. Mol. Biol., 1970, 50:279-294), then for a single cross-bridge the radial component of the cross-bridge force is not quite so sensitive to actin site location and is approximately 0.1 the axial component. In both cases, the ratio of the radial to axial force decreases with decreasing filament separation. If the radial-axial force ratio for each cross-bridge is approximately 0.1, then at full overlap in a frog skeletal muscle fiber the radial component of the cross-bridge force accompanying full activation will exert a compressive pressure of approximately 5 X 10(-3) atm. This would have little effect upon an intact muscle fiber where the volume constraints are likely osmotic, but it might produce a 1-2% change in filament spacing in a "skinned" muscle fiber from which the sarcolemma had been removed. These computations assume that the S2 link between the S1 head and the myosin filament does not support a bending moment of shear. If it does, then the radial component of the cross-bridge will be either greater or less, depending on the specific cross-bridge geometry.


Safety, effectiveness and mid-term follow-up in 136 consecutive patients with moderate to severely calcified lesions undergoing phoenix atherectomy.
To investigate the safety and effectiveness of the Phoenix atherectomy device for the treatment of complex and calcified lesions in patients with peripheral artery disease (PAD). 136 consecutive all-comer patients with chronic PAD underwent Phoenix atherectomy. Safety in terms of vessel injury and embolism, efficacy and clinical success in terms of ≥ 1Rutherford class (RF) improvement during follow-up were systematically analyzed. Lesion calcification was categorized by the Peripheral Arterial Calcium Scoring System (PACSS), whereas lesion complexity was classified by the Transatlantic Inter-Society Consensus (TASC). 151 lesions were treated in 136 consecutive patients. Clinical follow-up was available at 10.3 ± 4.2 months in 132 (97%) patients. 55 patients (40%) had intermittent claudication, 16 (12%) rest pain and 65 (48%) had ischemic ulcerations (mean RF class = 4.2 ± 1.1). 15 (11%) patients had TASC B lesions, whereas the majority 72 (53%) and 49 (36%) exhibited TASC C and D lesions, respectively. Mean PACSS score was 3.3 ± 0.9. Mean lesion length was 106 ± 92 mm. Atherectomy was combined with drug-coated balloon (DCB) in 129 (95%) patients. Nine (6.6%) patients with infra-inguinal lesions received stents. Technical and procedural success were recorded in 102 (75%) and 135 (99%), respectively. Perforation was noticed in 2 (1%), whereas asymptomatic embolism occurred in 6 (4%) patients. Clinical success was present in 54 (100%) patients with claudication and in 65 of 78 (83%) patients with critical limb ischemia (CLI). Atherectomy in combination with DCB angioplasty can be safely performed in patients with complex, calcified peripheral lesions with a relatively low rate of bail-out stenting and promising clinical mid-term results.German Clinical Trials Register: DRKS00016708.


Usefulness of an Implantable Loop Recorder to Detect Clinically Relevant Arrhythmias in Patients With Advanced Fabry Cardiomyopathy.
Patients with genetic cardiomyopathy that involves myocardial hypertrophy often develop clinically relevant arrhythmias that increase the risk of sudden death. Consequently, guidelines for medical device therapy were established for hypertrophic cardiomyopathy, but not for conditions with only anecdotal evidence of arrhythmias, like Fabry cardiomyopathy. Patients with Fabry cardiomyopathy progressively develop myocardial fibrosis, and sudden cardiac death occurs regularly. Because 24-hour Holter electrocardiograms (ECGs) might not detect clinically important arrhythmias, we tested an implanted loop recorder for continuous heart rhythm surveillance and determined its impact on therapy. This prospective study included 16 patients (12 men) with advanced Fabry cardiomyopathy, relevant hypertrophy, and replacement fibrosis in "loco typico." No patients previously exhibited clinically relevant arrhythmias on Holter ECGs. Patients received an implantable loop recorder and were prospectively followed with telemedicine for a median of 1.2 years (range 0.3 to 2.0 years). The primary end point was a clinically meaningful event, which required a therapy change, captured with the loop recorder. Patients submitted data regularly (14 ± 11 times per month). During follow-up, 21 events were detected (including 4 asystole, i.e., ECG pauses ≥3 seconds) and 7 bradycardia events; 5 episodes of intermittent atrial fibrillation (>3 minutes) and 5 episodes of ventricular tachycardia (3 sustained and 2 nonsustained). Subsequently, as defined in the primary end point, 15 events leaded to a change of therapy. These patients required therapy with a pacemaker or cardioverter-defibrillator implantation and/or anticoagulation therapy for atrial fibrillation. In conclusion, clinically relevant arrhythmias that require further device and/or medical therapy are often missed with Holter ECGs in patients with advanced stage Fabry cardiomyopathy, but they can be detected by telemonitoring with an implantable loop recorder.