Severe gastroparesis is associated with an increased incidence of slow-transit constipation as measured by wireless motility capsule.
Dysmotility in one region of the gastrointestinal tract has been found to predispose patients to developing motility disorders in other gastrointestinal segments. However, few studies have evaluated the relationship between gastroparesis and constipation. Retrospective review of 224 patients who completed 4-hour, solid-phase gastric emptying scintigraphy (GES), and wireless motility capsule (WMC) testing to evaluate for gastroparesis and slow-transit constipation, respectively. When available, anorectal manometry data were reviewed to evaluate for dyssynergic defecation. Patients were divided into two groups based on the results of the GES: 101 patients with normal gastric emptying and 123 patients with gastroparesis (stratified by severity). Differences in constipation rates were compared between the groups. Slow-transit constipation was more common in the gastroparesis group, but statistical significance was not reached (42.3% vs 34.7%, p = 0.304). Univariate logistical regression analysis found no association between slow-transit constipation and gastroparesis (OR 1.38, 95% CI 0.80-2.38, p = 0.245) nor dyssynergic defecation and gastroparesis (OR 0.88, 95% CI 0.29-2.70, p = 0.822). However, when stratifying gastroparesis based on severity, slow-transit constipation was found to be associated with severe gastroparesis (OR 2.45, 95% CI 1.20-5.00, p = 0.014). This association was strengthened with the exclusion of patients with diabetes mellitus (OR 3.5, 95% CI 1.39-8.83, p = 0.008) - a potential confounder. Patients with severe gastroparesis (>35% gastric retention at the 4-hour mark on solid-phase GES) have an increased likelihood of having underlying slow-transit constipation. Dyssynergic defecation does not appear to be associated with gastroparesis (of any severity).


Quantitative Analysis of STD-NMR Spectra of Reversibly Forming Ligand-Receptor Complexes.
We describe our work on the quantitative analysis of STD-NMR spectra of reversibly forming ligand-receptorcomplexes. This analysis is based on the theory of complete relaxation and conformational exchange matrixanalysis of saturation transfer (CORCEMA-ST) effects. As part of this work, we have developed two separateversions of the CORCEMA-ST program. The first version predicts the expected STD intensities for a givenmodel of a ligand-protein complex, and compares them quantitatively with the experimental data.This version is very useful for rapidly determining if a model for a given ligand-proteincomplex is compatible with the STD-NMR data obtained in solution. It is also useful in determining theoptimal experimental conditions for undertaking the STD-NMR measurements on a given complex by computersimulations. In the second version of the CORCEMA-ST program, we have implemented a torsion anglerefinement feature for the bound ligand within the protein binding pocket. In this approach, the globalminimum for the bound ligand conformation is obtained by a hybrid structure refinement protocol involvingCORCEMA-ST calculation of intensities and simulated annealing refinement of torsion angles of the boundligand using STD-NMR intensities as experimental constraints to minimize a pseudo-energy function.This procedure is useful in refining and improving the initial models based on crystallography, computerdocking, or other procedures to generate models for the bound ligand within the protein binding pocket compatiblewith solution STD-NMR data. In this chapter we describe the properties of the STD-NMR spectra, includingthe dependence of the intensities on various parameters. We also describe the results of the CORCEMA-STanalyses of experimental STD-NMR data on some ligand-protein complexes to illustrate the quantitativeanalysis of the data using this method. This CORCEMA-ST program is likely to be useful in structure-baseddrug design efforts.


Fatty acid binding protein 4 predicts left ventricular mass and longitudinal function in overweight and obese women.
To explore whether increased adipocyte-derived serum fatty acid binding protein 4 (FABP4) predisposes to cardiac remodelling and left ventricular dysfunction in human obesity. Cross-sectional investigation. Academic clinical research centre. 108 overweight and obese non-diabetic women (body-mass index 33 ± 5 kg/m2). None. Relationship between serum FABP4 and abdominal adipose tissue quantified by MRI. Relationship between serum FABP4 and left ventricular morphology and function assessed by cardiac MRI. FABP4 was independently associated with visceral abdominal adipose tissue (β=0.34, p<0.01) and subcutaneous abdominal adipose tissue (β=0.22, p<0.05). After stratification into serum FABP4 tertiles, left ventricular masses were 92 ± 16 g, 86 ± 13 g and 81 ± 12 g in women with high, intermediate and low FABP4 concentrations (p<0.01), respectively. Longitudinal systolic function was reduced by 8% in women with intermediate and high versus low FABP4 concentrations (p<0.01), whereas ejection fraction did not differ among tertiles (p=0.5). In multivariate linear analysis FABP4 remained an independent predictor of left ventricular mass (β=0.17, p<0.05) and reduced longitudinal fractional shortening (β=0.21, p<0.05). In overweight and obese women, FABP4 showed an independent association with parameters of left ventricular remodelling.


Expression of eotaxin, an eosinophil-selective chemokine, parallels eosinophil accumulation in the vesiculobullous stage of incontinentia pigmenti.
Incontinentia pigmenti (IP) is an X-linked dominant genodermatosis primarily affecting female children. The initial vesiculobullous stage of IP is characterized clinically by inflammatory papules, blisters, and pustules, and histopathologically by acanthosis, keratinocyte necrosis, epidermal spongiosis and massive epidermal eosinophil infiltration. The cause of this multisystem disease is attributed to the mutations of an X-linked regulatory gene, termed nuclear factor-kappaB essential modulator (NEMO). The exact mechanism of epidermal eosinophil accumulation has not yet been determined. We explored the possible role of an eosinophil-selective, nuclear factor-kappaB-activated chemokine, eotaxin, in the accumulation of eosinophils in the initial stage of the disease. Monoclonal antibody (6H9) specific for human eotaxin strongly labelled the suprabasal epidermis of IP skin, paralleling the upper epidermal accumulation of eosinophils, but did not label the epidermis of normal skin or lesional skin from patients with other inflammatory skin diseases not characterized by prominent eosinophil accumulation, namely dermatitis herpetiformis and selected cases of atopic dermatitis lacking significant numbers of eosinophils. In addition, endothelial cells in lesional skin of IP also exhibited strong expression of eotaxin, which correlated with perivascular and intravascular eosinophil infiltration. We also examined the in vitro effects on epidermally derived eotaxin of several cytokines that were nuclear factor-kappaB-activated and/or known to induce eotaxin expression. In normal human keratinocytes, proinflammatory cytokines either independently (IL-1alpha) or synergistically (tumour necrosis factor-alpha (TNF-alpha)/ interferon-gamma (IFN-gamma) and TNF-alpha/IL-4) up-regulated eotaxin expression. These studies suggest that release of cytokines during the initial inflammatory stage of IP induces epidermal expression of eotaxin, which may play a role in the epidermal accumulation of eosinophils.


Temporal effects of tumor necrosis factor-alpha on intracellular survival of Mycobacterium paratuberculosis.
The causative agent in Johne's disease is Mycobacterium paratuberculosis, an intracellular pathogen which causes enteritis in ruminants. Little is known about interactions between the host cell (macrophage) and M. paratuberculosis; however, this bacterium is able to evade normal host immune defenses and cause a chronic infective state. In the present study, we evaluated whether activation of a murine macrophage cell line (J774.16) by pretreatment with recombinant murine tumor necrosis factor alpha (TNF) prior to infection with M. paratuberculosis would affect their ability to restrict growth and kill the ingested bacteria. A murine cell line was utilized owing to difficulty in obtaining bovine reagents and lack of a continuous bovine macrophage cell line for repeated experimentation. After 4 h of infection, numbers of viable bacteria in cell lysates were significantly lower for macrophages pretreated with 1000 IU TNF ml-1. The rate of bacterial growth as assessed by BACTEC radiometric culture system was also reduced at this time point. Upon further extension of the infection period to 72 h, we observed that moderate doses of TNF (10-1000 IU ml-1) significantly increased the number of viable M. paratuberculosis recovered whereas the highest dose of TNF (4000 IU ml-1) effectively reduced bacterial numbers. These data indicate that TNF can either enhance or reduce macrophage mycobactericidal and mycobacteriostatic activity depending upon both the level of TNF to which cells are exposed and the duration of infection.


Stimulation of extramedullary hemopoiesis by dextran sulfate.
The spleen of normal dogs (beagles) shows only slight hemopoietic activity, characterized by the presence of megakaryocytes in mitosis and small groups of erythroblasts scattered throughout the red pulp of the organ. Repeated intravenous injection of dextran sulfate, at a dose of 15 mg per kg body weight, produced markedly enhanced erythropoietic and megakaryocytopoietic activity in the splenic red pulp, without concomitant increase in splenic granulopoiesis. The probable existence of a micro-environment adequate for erythro- and megakaryocytopoietic differentiation of stem cells is discussed.


Molecular and epidemiological study of enterovirus D68 in Taiwan.
As an immunofluorescence assay for enterovirus D68 (EV-D68) is not available in the enteroviruses surveillance network in Taiwan, EV-D68 may be the actual pathogen of untypeable enterovirus-suspected isolates. The untypeable isolates collected from 2007 through 2014 were identified by nucleic acid amplification-based methods and sequencing of the VP1 region to analyze the phylogeny and epidemiology of EV-D68 in Taiwan. Twenty-nine EV-D68 isolates were sequenced, including 15 Cluster 3 and 14 Cluster 1 viruses. Approximately 41% of the patients were children under 5 years of age and their infections peaked in August. The ratio of male to female patients was 1.5 and 3.67 for Cluster 3 and Cluster 1, respectively. Fever and respiratory symptoms were commonly reported in EV-D68-infected patients. The results of phylogenetic analyses showed that EV-D68 isolates between 2007 and 2014 belonged to different clusters and existed for years, indicating that endemic circulation of EV-D68 existed in Taiwan. This study showed that EV-D68 has been endemic in Taiwan for some years despite a small number of positive cases. The continuous monitoring and efforts towards the improvement of diagnostic techniques are required to complete the surveillance system. This study provided the genetic and epidemiological information which could contribute to understanding the etiology and epidemiology of EV-D68.


Surfactant protein A gene deletion and prognostics for patients with stage I non-small cell lung cancer.
The present study was conducted to determine clinical relevance of surfactant protein A (SP-A) genetic aberrations in early-stage non-small cell lung cancer (NSCLC). To determine whether SP-A aberrations are lung cancer-specific and indicate smoking-related damage, tricolor fluorescence in situ hybridization with SP-A and PTEN probes was done on touch imprints from the lung tumors obtained prospectively from 28 patients with primary NSCLC. To further define the clinical relevance of SP-A aberrations, fluorescence in situ hybridization was done on both tumor cells and adjacent bronchial tissue cells from paraffin-embedded tissue blocks from 130 patients NSCLC for whom we had follow-up information. SP-A was deleted from 89% of cancer tissues and the deletion was related to the smoking status of patients (P < 0.001). PTEN was deleted from 16% in the cancer tissues and the deletion was not related to the smoking status of patients (P > 0.05). In the cells isolated from paraffin-embedded tissue blocks, SP-A was deleted from 87% of the carcinoma tissues and 32% of the adjacent normal-appearing bronchial tissues. SP-A deletions in tumors and adjacent normal-appearing bronchial tissues were associated with increases in the risk of disease relapse (P = 0.0035 and P < 0.001, respectively). SP-A deletions in the bronchial epithelium were the strongest prognostic indicators of disease-specific survival (P = 0.025). Deletions of the SP-A gene are specific genomic aberrations in bronchial epithelial cells adjacent to and within NSCLC, and are associated with tumor progression and a history of smoking. SP-A deletions might be a useful biomarker to identify poor prognoses in patients with NSCLC who might therefore benefit from adjuvant treatment.


The impact of Hospital Elder Life Program interventions, on 30-day readmission Rates of older hospitalized patients.
This study investigated the impact of the Hospital Elder Life Program (HELP), a geriatric intervention program targeted at reducing delirium, on readmission rates. Secondary goals were to review HELP's impact on average length of stay (ALOS), discharge to the home, and falls. This retrospective 3-year study reviewed inpatients age 70 and older, with one or more of six impairments: functional, renal, cognitive, sleep, visual and hearing. Patients were from a 784 bed teaching hospital in California with admissions between January 1, 2014 and December 31, 2016. This was compared to a matched cohort of patients who did not receive any interventions from HELP volunteers or a HELP geriatric nurse practitioner. Data was also reviewed for impact on goals in age groups 70-85 and 85 + . 2146 HELP patients with an average age of 82.1 showed improvement in 30-day readmission rates, with a readmission rate of 11.3% in the HELP group and 13.5% in the control group (n = 2704, mean 82.5). Detailed age data noted statistical significance only in the 70-85 population for readmission. ALOS was 5.6 and 6.2 days respectively in the HELP and non-HELP population. ALOS was also statistically significant across 70-85 and 85 + . Neither rates of discharge to home nor falls showed any significant improvement. Both control and intervention groups had a disproportionate number of whites and females. This study shows hospitals can improve patient outcomes and financial burden from 30-day readmissions and length of stay, particularly in the 70-85 age group, by implementing the Hospital Elder Life Program.


Trauma: when there's no time to count.
Trauma is the leading cause of death for people younger than age 45 in the United States. To help prevent death or disability, critically injured trauma patients must reach definitive care within the "golden hour."Often, the perioperative team has no more than 10 minutes to prepare the room before a patient who has suffered multiple, critical, traumatic injuries is rushed into the OR. For the trauma patient's safety, the perioperative team must be diligent and use creative measures to efficiently ensure that all surgical counts are completed. Foregoing surgical counts places the trauma patient at an increased risk for unintentionally retained foreign objects.


Annexin A2 regulates unfolded protein response via IRE1-XBP1 axis in macrophages during P. aeruginosa infection.
Pseudomonas aeruginosa is a severe Gram-negative opportunistic bacterium that causes a spectrum of organ system diseases, particularly in immunocompromised patients. This bacterium has been shown to induce unfolded protein response (UPR) during mammalian infection. Annexin A2 (AnxA2) is a multicompartmental protein relating to a number of cellular processes; however, it remains unknown whether AnxA2 coordinates a UPR pathway under bacterial infection conditions. Here, we report that the endoplasmic reticulum stress inositol-requiring enzyme 1 (IRE1)-X-box binding protein 1 (XBP1) pathway was up-regulated by AnxA2 through p38 MAPK signaling following P. aeruginosa infection in macrophages, whereas ATF4 and ATF6 not. In addition, XBP1 was found as a positive regulator of innate immunity to tame P. aeruginosa challenges by enhancing autophagy and bacterial clearance. XBP1 also facilitated NF-κB activation to elicit the release of proinflammatory cytokines predominantly in macrophages. Together, our findings identify AnxA2 as a regulator for XBP1-mediated UPR pathway.


The failure of polypropylene surgical mesh in vivo.
Surgical mesh materials made from fibres of polypropylene (PP) have been widely used for over fifty years. However in recent times the use of these materials has been called into question for certain surgical operations, known as "pelvic organ" or "transvaginal" procedures. This article reviews the current state of knowledge and uses some simple biomechanics analysis in order to make recommendations for future work. My conclusion is that the failure rate of PP mesh in pelvic organ products is unacceptably high whilst being much lower when it is used for hernia repair. The precise mechanical environment is difficult to quantify but a tentative conclusion is that failure by purely mechanical mechanisms (fatigue and creep) is unlikely. There is some evidence for environmental degradation (i.e. stress-corrosion) but this is highly contested in the literature. On balance my conclusion is that stress-corrosion failure is the most likely explanation but currently no experimental results exist to prove conclusively that this failure mechanism occurs in vivo. Further work is needed, especially the analysis of explanted material, to resolve this urgent problem.


Standard in vitro evaluations of engineered bone substitutes are not sufficient to predict in vivo preclinical model outcomes.
Understanding the optimal conditions required for bone healing can have a substantial impact to target the problem of non-unions and large bone defects. The combination of bioactive factors, regenerative progenitor cells and biomaterials to form a tissue engineered (TE) complex is a promising solution but translation to the clinic has been slow. We hypothesized the typical material testing algorithm used is insufficient and leads to materials being mischaracterized as promising. In the first part of this study, human bone marrow - derived mesenchymal stromal cells (hBM-MSCs) were embedded in three commonly used biomaterials (hyaluronic acid methacrylate, gelatin methacrylate and fibrin) and combined with relevant bioactive osteogenesis factors (dexamethasone microparticles and polyphosphate nanoparticles) to form a TE construct that underwent in vitro osteogenic differentiation for 28 days. Gene expression of relevant transcription factors and osteogenic markers, and von Kossa staining were performed. In the second and third part of this study, the same combination of TE constructs were implanted subcutaneously (cell containing) in T cell-deficient athymic Crl:NIH-Foxn1rnu rats for 8 weeks or cell free in an immunocompetent New Zealand white rabbit calvarial model for 6 weeks, respectively. Osteogenic performance was investigated via MicroCT imaging and histology staining. The in vitro study showed enhanced upregulation of relevant genes and significant mineral deposition within the three biomaterials, generally considered as a positive result. Subcutaneous implantation indicates none to minor ectopic bone formation. No enhanced calvarial bone healing was detected in implanted biomaterials compared to the empty defect. The reasons for the poor correlation of in vitro and in vivo outcomes are unclear and needs further investigation. This study highlights the discrepancy between in vitro and in vivo outcomes, demonstrating that in vitro data should be interpreted with extreme caution. In vitro models with higher complexity are necessary to increase value for translational studies. STATEMENT OF SIGNIFICANCE: Preclinical testing of newly developed biomaterials is a crucial element of the development cycle. Despite this, there is still significant discrepancy between in vitro and in vivo test results. Within this study we investigate multiple combinations of materials and osteogenic stimulants and demonstrate a poor correlation between the in vitro and in vivo data. We propose rationale for why this may be the case and suggest a modified testing algorithm.


Data on identification of conserved and novel miRNAs in Elettaria cardamomum.
Elettaria cardamomum (L.) Maton, or small cardamom referred as 'queen of spices', is a perennial herbaceous rhizomatous monocot of the family Zingiberaceae. Cardamom seeds and fruits are the economically significant parts and effectively used as a traditional medicine, food additive and flavoring agent. In the present study, using Ion Proton next generation sequencing technology we performed the small RNA sequencing, conserved and novel miRNA predictions of a wild and five cultivar genotypes of cardamom. Small RNA sequencing generated a total of 5,451,328 and 2,756,250 raw reads for wild and cultivar cardamom respectively. The raw data was submitted to SRA database of NCBI under the accession numbers and SRX2273863 (wild) and SRX2273862 (cultivars). The raw reads were quality filtered and predicted conserved and novel miRNAs for wild and cultivar cardamom. The predicted miRNAs, miRNA-targets and functional annotations might provide valuable insights into differences between wild progenitor and cultivated cardamom.


Knowledge and attitude about pubertal health and their socio-demographic predictors in Iranian adolescents.
Adolescence is a critical period and is associated with physical and psycho-social changes induced by puberty, which builds personality, behavior, and future health of the adolescents. This study aimed to determine the adolescent's knowledge and attitude toward pubertal health and his/her socio-demographic predictors. This cross-sectional study was performed on 1017 adolescent girls who were in the second and third grades of secondary schools in Tabriz, Iran, in 2013. The sampling method was random. The general linear model was used to determine the social-demographic predictors of the adolescent's knowledge and attitude about pubertal health. The mean (standard deviation) score of knowledge and attitude was 53.2 (13.7) and 66.7 (11.5), respectively (from an attainable score range of 0-100). The variables of educational level, economic status, household members, obtaining information about the puberty, and age of menarche were predictors of knowledge. Educational level, the most feeling at menarche, and source of the most information about puberty were predictors of attitude (p<0.05). The findings of this study show that the knowledge and attitude of adolescents about pubertal health are moderate. Thus, it is necessary to educate the girls about pubertal health to enable them to have a healthy reproductive life in the future.


Reporting of research data by GPs: a cautionary tale for primary care researchers.
Given the importance of community-based research, there is a need to ensure the quality of data obtained from such studies. However, research has been considered a low priority for most GPs. To assess the quality of data reported by GPs in a large community-based study. Men were recruited as part of a population-based study on prostate-specific antigen (PSA) testing. Those with elevated initial PSA levels in the mid-1990s and no investigation in the intervening period were invited for repeat PSA testing. The GP of each included man was contacted and asked to report basic clinical information. Trained data extractors independently reviewed each man's GP medical notes and recorded data relating to prostate problems. Data provided by GPs were rematched with data extracted from medical notes. A total of 758 men with 379 GPs were included. In all, 366 (96.6%) GPs agreed to participate. Of 698 men suitable for follow-up, GP and note review data were available in 505 (72.3%) cases (287 GPs). Overall, 245 (85.4%) GPs provided completely accurate data. Male GPs, compared to female GPs, were found to have a higher level of inaccurate reporting (P = 0.008). We found that when GPs were asked to record basic clinical information, for the purposes of a primary care-based study, there was a significant level of inaccurate reporting. The results from our study confirm the importance of quality control in primary care research, especially in studies that involve GPs' reporting data.


Aldehyde dehydrogenase activity as a functional marker for lung cancer.
Aldehyde dehydrogenase (ALDH) activity has been implicated in multiple biological and biochemical pathways and has been used to identify potential cancer stem cells. Our main hypothesis is that ALDH activity may be a lung cancer stem cell marker. Using flow cytometry, we sorted cells with bright (ALDH(br)) and dim (ALDH(lo)) ALDH activity found in H522 lung cancer cell line. We used in vitro proliferation and colony assays as well as a xenograft animal model to test our hypothesis. Cytogenetic analysis demonstrated that the ALDH(br) cells are indeed a different clone, but when left in normal culture conditions will give rise to ALDH(lo) cells. Furthermore, the ALDH(br) cells grow slower, have low clonal efficiency, and give rise to morphologically distinct colonies. The ability to form primary xenografts in NOD/SCID mice by ALDH(br) and ALDH(lo) cells was tested by injecting single cell suspension under the skin in each flank of same animal. Tumor size was calculated weekly. ALDH1A1 and ALDH3A1 immunohistochemistry (IHC) was performed on excised tumors. These tumors were also used to re-establish cell suspension, measure ALDH activity, and re-injection for secondary and tertiary transplants. The results indicate that both cell types can form tumors but the ones from ALDH(br) cells grew much slower in primary recipient mice. Histologically, there was no significant difference in the expression of ALDH in primary tumors originating from ALDH(br) or ALDH(lo) cells. Secondary and tertiary xenografts originating from ALDH(br) grew faster and bigger than those formed by ALDH(lo) cells. In conclusion, ALDH(br) cells may have some of the traditional features of stem cells in terms of being mostly dormant and slow to divide, but require support of other cells (ALDH(lo)) to sustain tumor growth. These observations and the known role of ALDH in drug resistance may have significant therapeutic implications in the treatment of lung cancer.


[The Leydig-cell testicular tumor. A case report with magnetic resonance images].
An additional case of testicular Leydig cell tumor in a young man is reported. The clinical presentation included gynecomastia and a testicular mass. Hormonal studies, testicular US and high resolution MRI were performed before orchidectomy. The histopathological findings were consistent with Leydig cell tumor. A CT scan revealed no metastasis. The patient is clinically disease-free 18 months after orchidectomy. High resolution MRI must be performed only when clinic and US findings are discordant.


A transducer for detection of fetal breathing movements.
The paper describes a new transducer for detection of fetal movements and sounds from the maternal abdominal wall. This transducer is based on an inductive principle. The compliance of the transducer should match the compliance of the maternal abdominal wall in order to detect the very weak acoustic signal caused by individual fetal breathing movements. The contact area of the transducer is supported on a membrane, the tension of which can be adjusted so as to match the compliances of tissue and transducer. The goal of this paper is to investigate detection of fetal movements in order to monitor fetal condition and motility. The more specialized field of investigating fetal heart rate variability by studying the relationship between fetal breathing movements and fetal heart rate may become of more interest in the future. The inductive phonometer (INPHO) shows a flat (+/- 1.5 dB) frequency response between 0.2 and 200 Hz which is not affected by the compliance adjustment. Some in vivo measurements have been performed. The transducer signal is stored on a 20MB hard disk of an Olivetti M24 Personal Computer (PC) simultaneously with a control signal indicating the presence of breathing movements. Ultrasound imaging is exclusively used for verification of breathing movements. The images are stored on videotape simultaneously with control signals from the PC for synchronization of data and image. Substantial digital filtering is necessary to discriminate between actual breathing movements and other fetal activities. It is shown that optimal adjustment of the transducer's compliance to that of the maternal abdominal wall is possible and that individual breathing movements can indeed be measured.


Severe hemolysis after incomplete mitral valve repair.
We report 2 cases of severe intravascular hemolysis after mitral valve repair using a Duran annuloplasty ring. In both patients residual mitral regurgitation was present, hemolysis was severe enough to warrant a second operation, and hemolysis ceased immediately after the second operation. We believe that a high-velocity regurgitant jet directed toward the cloth-covered annuloplasty ring was responsible for the hemolysis in both patients.


Effectively Communicating Colorectal Cancer Screening Information to Primary Care Providers: Application for State, Tribe or Territory Comprehensive Cancer Control Coalitions.
Patients are more likely to be screened for colorectal cancer if it is recommended by a health care provider. Therefore, it is imperative that providers have access to the latest screening guidelines. This practice-based project sought to identify Kentucky primary care providers' preferred sources and methods of receiving colorectal cancer information to improve state comprehensive cancer control provider outreach initiatives. Four focus groups were conducted with primary care physicians, nurse practitioners, and physician assistants. Discussion included preferred sources and methods of receiving updated screening guidelines, legislation, and statewide public awareness campaign materials. Providers (N = 17) identified their preferred methods for receiving colorectal cancer information as: routine emails from trusted sources (colleagues, professional societies and research, and advocacy agencies), scientific journals, existing conferences, and the media. When delivering colorectal cancer information to primary care providers, multiple approaches are needed. An ideal partner for dissemination of information is state comprehensive cancer control coalitions, considering their prioritization of colorectal cancer screening and existing networks of partners who were identified as trusted sources. Assessment of primary care providers' preferred methods and sources of receiving colorectal cancer information informs strategies for practice among comprehensive cancer control coalitions.


Social determinants and spatio-temporal variation of Ischemic Heart Disease in Manitoba.
The aim was to study any spatial and/or temporal patterns of ischemic heart disease (IHD) prevalence and measure the effects of selected social determinants on these spatial and space-time patterns. Data were obtained from the Population Research Data Repository housed at the Manitoba Centre for Health Policy to identify persons who were diagnosed with IHD between 1995 and 2018. These persons were geocoded to 96 geographic regions of Manitoba. An area-level socioeconomic factor index (SEFI-2) and the proportion of the population who was Indigenous were calculated for each geographic region using the 2016 Canadian Census data. Associations between these factors and IHD prevalence were measured using Bayesian spatial Poisson regression models. Temporal trends and spatio-temporal trends were measured using Bayesian spatio-temporal Poisson regression models. Univariable models showed a significant association with increased regional Indigenous population proportion associated with a higher prevalence of IHD (RR: 0.07, 95% CredInt: (0.05, 0.10)) and for SEFI-2 (RR: 0.17, 95% CredInt: (0.11, 0.23)). Using a multivariable model, after accounting for the proportion of the population that was Indigenous, there was no evidence of an association between IHD prevalence and area-level socioeconomic factor. Spatio-temporal models showed no significant overall temporal trend in IHD prevalence, but there were significant spatially varying temporal trends within the 96 regions. Association between Indigenous population proportion and IHD is consistent with previous research. No significant overall temporal trend was measured. However, regions with significantly increasing trends and significantly decreasing trends in IHD prevalence were identified.


Artifacts in Contrast-Enhanced Ultrasound during Follow-up after Endovascular Aortic Repair: Impact on Endoleak Detection in Comparison with Computed Tomography Angiography.
The study described here systematically analyzed how specific artifacts in contrast-enhanced ultrasound (CEUS) can affect the detection of endoleaks during follow-up after endovascular aortic repair (EVAR). Patients undergoing EVAR of atherosclerotic or mycotic abdominal aortic aneurysms using various standard and branched stent-graft material for visceral and iliac preservation were enrolled over 5 y and followed up with computed tomography angiography (CTA) and CEUS simultaneously. CEUS artifacts were frequently identified after EVAR procedures (59% of examinations) and were caused mainly by contrast agent, different prosthesis or embolization material and postinterventional changes in the aneurysm sac. This article describes how to identify important artifacts and how to avoid false-negative or false-positive interpretations of endoleaks. Despite artifacts, CEUS had higher sensitivity for endoleak detection after EVAR than CTA. CEUS was superior to CTA in the identification of late endoleaks type II and in follow-up examinations after embolization procedures, where beam-hardening artifacts limited CTA.


Advancing sensing technology with CRISPR: From the detection of nucleic acids to a broad range of analytes - A review.
The CRISPR/Cas technology, derived from an adaptive immune system in bacteria, has been awarded the Nobel Prize in Chemistry in 2020 for its success in gene editing. Increasing reports reveal that CRISPR/Cas technology has a wide scope of applications and it could be incorporated into biosensors for detecting critical analytes. CRISPR-powered biosensors have attracted significant research interest due to their advantages including high accuracy, good specificity, rapid response, and superior integrity. Now the CRISPR technology is not only admirable in nucleic acid monitoring, but also promising for other kinds of biomarkers' detection, including metal ions, small molecules, peptides, and proteins. Therefore, it is of great worth to explore the prospect, and summarize the strategies in applying CRISPR technology for the recognition of a broad range of targets. In this review, we summarized the strategies of CRISPR biosensing for non-nucleic-acid analytes, the latest development of nucleic acid detection, and proposed the challenges and outlook of CRISPR-powered biosensors.


High-stiffness distal fixation restores anterior laxity and stiffness as well as joint line fixation with an interference screw.
Fixation of an anterior cruciate ligament graft at the joint line with an interference screw restores anterior laxity better than low-stiffness distal fixation with double staples and sutures tied to a post at implantation in porcine bone. Fixation of an anterior cruciate ligament graft with a high-stiffness distal fixation device restores anterior laxity and knee stiffness as well as joint line fixation with an interference screw in human bone. Controlled laboratory study. Eleven cadaveric knees with foam reinforcement of the bone were reconstructed with a double-looped tendon graft and fixed with an interference screw and 3 high-stiffness methods of fixation (tandem washer, WasherLoc, and WasherLoc with bone dowel). Anterior laxity at 225 and 110 N of anterior load and knee stiffness were measured at 0 degrees , 30 degrees , 60 degrees , 90 degrees , and 120 degrees of flexion with a 6 degrees of freedom load application system. The increase in anterior laxity and knee stiffness with the interference screw was not significantly different from that with the high-stiffness distal methods. In the human knee, the use of high-stiffness fixation devices placed distal to the joint line restored anterior laxity and knee stiffness as well as joint line fixation with an interference screw at implantation. The stiffness provided by each method of fixation should be considered along with the length of the graft when determining anterior laxity and knee stiffness.


Renal prostaglandins.
Relative renal ischemia, produced by a variety of interventions, is modulated or buffered by prostaglandins synthesized within the kidney. These prostaglandins, however, do not account for renal autoregulation in its classic sense. They can facilitate salt and water excretion, largely through effects on physical forces in proximal peritubular capillaries, as evidenced by augmented renal blood flow. Renal prostaglandins may play a role in chronic water balance, but they probably are not important in chronic electrolyte homeostasis. There are several potential mechanisms by which renal prostaglandins could exert antihypertensive efects. Establishment of the importance of these agents, however, as antihypertensive hormones remains a challenge for future investigation.


Risk factors, classification, and operative choices of femur fractures at a Tertiary Hospital: first report from Somalia.
A traumatic femur fracture is a significant cause of morbidity, affecting one to three million individuals annually. The present is the first study investigated the epidemiological characteristics, risk factors, classification, mechanisms of injury, and early management of femoral fractures in Somalia. This retrospective epidemiological study included all patients with a femur fracture who were admitted for four years between November 2018 and December 2022 to the orthopedic and trauma surgery department. We reviewed patient demographic characteristics, including age and gender, the mechanism of injury, injury characteristics, and the type of fixation performed. We reviewed the radiographs and classified the fracture using the AO/OTA classification system. During the study period, a total of 402 patients were treated for femur fractures; 256 (64%) were males, and 144 (36%) were females. The mean patient age was 47.7 ± 8.5 years. Regarding the anatomical location of femur fractures, the proximal (31A, 31B) was the most common, accounting for 50% of the patients. Femur neck fracture (31B) was the most common in the proximal femur fractures. Gunshot 82 (59.42%) was the leading cause of femur shaft fractures. Most patients with femur shaft fractures were males; 150 (86.20%) and 152 (64.47%) were young patients between 19 and 40 years old. Almost half of the patients (86) with femur shaft fractures had open fractures. The distribution of the mechanism of injury significantly differed according to age (p < 0.001). Younger patients (< 40 years) were predominantly injured due to gunshot injuries compared to elderly cases (> 60 years), where falls from standing height were the primary mechanism of injuries. There was a statistically significant difference between the mechanism of injury and gender categories (p < 0.001). Male patients were injured mainly by gunshots in about 40%, while 80% of fractures in female patients were due to falls from standing height. Female fractures occurred primarily in the proximal, while the males had an equal fracture rate for proximal and shaft fractures. Femur fracture causes significant morbidity and mortality. The study findings revealed that the most common femur fracture type was femur neck fracture, and low-energy injuries were the most common mode of injury in the elderly. Proximal femur fractures occur in older age and mainly in females. Gunshots were the most common cause of femur shaft fractures in Somalia, a country that has struggled with wars for over 30 years.


Development and characterization of insect cell lines.
With the wide availability of insect cell culture media, it can generally be considered a routine process to develop new cell lines. Exceptions to this statement do exist, of course. Difficulties may arise when attempting to culture a specific cell type. For example, while there are a few cell lines from insect fat body and at least one from the midgut, it may not be possible to obtain cell lines from these tissues from all insect species due to terminal differentiation and other factors. Also, researchers have desired cell lines from certain species, such as the honey bee, for which no success has been obtained. As in the early days of tissue culture, it is difficult to discern why negative results occur. However, as more is learned about the physiology and nutrition of various insects and tissues, we may get clues which will help solve these questions.The remaining chapters in this book will provide the reader with exciting uses for insect cell culture. As I mentioned earlier, the baculovirus expression vector system has provided a stimulus to the field of insect cell culture not seen previously.


Durable Results of Mitomycin C Injection with Internal Urethrotomy for Refractory Bladder Neck Contractures: Multi-institutional Experience.
We evaluated our intermediate term experience with radial urethrotomy and intralesional mitomycin C injection in patients with recurrent bladder neck contractures. Recurrent bladder neck contractures in which previous endoscopic treatment failed pose a difficult management dilemma. Prospectively collected data were reviewed in a retrospective manner of patients presenting with recurrent bladder neck contractures from January 2007 to June 2014. All patients had at least 1 prior failed incision of a bladder neck contracture and many had additional dilations or catheter dependence. Radial cold knife incisions of the bladder neck were performed followed by injection of 0.3 to 0.4 mg/ml mitomycin C at each incision site. All surgeons performed the incision technique and injection in a reproducible fashion. A total of 40 patients underwent urethrotomy with mitomycin C injection. At a median followup of 20.5 months 30 patients (75.0%) had a stable bladder neck after 1 procedure. An additional 5 patients required 2 procedures to obtain a stable patent bladder neck (87.5%). Of the 40 patients 14 (35.0%) presented in retention on catheter drainage and all had a stable, patent bladder neck. No recurrence was detected in the original 18 patients in the pilot study with patent bladder necks. Rigorous followup revealed no long-term complications. Urethrotomy with mitomycin C injection for the management of recurrent bladder neck contractures is safe and efficacious. The addition of an antifibrotic agent in conjunction with internal urethrotomy offers a definitive solution to a problem that would otherwise be managed with repeat urethral incision/dilation, catheter dependence or open bladder neck reconstruction.


ErbB-beta-catenin complexes are associated with human infiltrating ductal breast and murine mammary tumor virus (MMTV)-Wnt-1 and MMTV-c-Neu transgenic carcinomas.
Simultaneous deregulation of both Wnt and ErbB growth factors has previously been shown to result in the cooperative induction of mammary gland tumors. Using the murine mammary tumor virus (MMTV)-Wnt-1 transgenic model of mammary carcinoma, we have identified an unvarying association between beta-catenin and epidermal growth factor receptor/c-Neu (ErbB1/ErbB2) heterodimers in mammary gland tumors, indicating a requirement for ErbB signaling in Wnt-mediated tumorigenesis. Expansion of these observations to a second transgenic model, MMTV-c-Neu, demonstrated similar tumor-specific interactions, including an ErbB1 ligand-inducible phosphorylation of both beta-catenin and c-Neu. Direct relevance of these findings to human breast cancer was established upon examination of a set of human infiltrating ductal breast adenocarcinoma and lymph node metastasis tissues taken at surgery. These data revealed increased levels of beta-catenin in tumors and metastases versus normal breast as well as an association between beta-catenin and c-Neu that measurably occurs only in neoplasia, most strongly in metastatic lesions. These studies have identified a seemingly indispensable interaction between beta-catenin and epidermal growth factor receptor/c-Neu heterodimers in Wnt-1-mediated breast tumorigenesis that may indicate a fundamental signaling event in human metastatic progression.


Maxillary defects due to COVID-19 associated mucormycosis: Impact on quality of life after rehabilitation with an obturator.
Aggressive surgical debridement including maxillectomy in patients with coronavirus disease 2019 (COVID-19) associated mucormycosis of the maxilla alters facial appearance, function, and an individual's overall quality of life (QOL). These patients require rehabilitation of defects with obturators to help them recover from esthetic and functional disabilities and regain near-normal quality of life. Because COVID-19 associated with mucormycosis is a newer entity, less is known about the quality of life of these patients after rehabilitation with obturators. Hence, the effects of COVID-19 associated with mucormycosis, treatment, and rehabilitation need to be evaluated to assess QOL. The purpose of this clinical study was to assess the impact on QOL after rehabilitation with obturators for participants with COVID-19-associated mucormycosis who underwent maxillectomy. Oral health impact profile-14 (OHIP-14) and obturator functioning scale (OFS) questionnaires were administered to 26 participants with COVID-19-associated mucormycosis of the maxilla who underwent maxillectomy and required rehabilitation with obturators. Questionnaire responses were received on numerical Likert scales of 0 to 4 for OHIP-14 and 1 to 5 for OFS. The repeated measures analysis of variance (ANOVA) with pairwise post hoc Bonferroni tests were used to evaluate and compare mean scores of OHIP-14 and OFS at different preoperative and postoperative stages (α=.05). A total of 26 participants who underwent maxillectomy were provided with obturators. A higher mean ±standard deviation OHIP score was observed before rehabilitation at T1 (51.0 ±9.6) followed by 1 week after fabrication of the surgical obturator at T2 (18.6 ±16.4), and 1 week after fabrication of the interim obturator at T3 (4.7 ±8.2). The lower mean scores ±standard deviation were seen 1 week after the fabrication of definitive obturators at T4 (2.9 ±7.2). Comparison of the scores at T1, T2, T3, and T4 found significant difference (P≤.001). Similarly, higher mean ±standard deviation OFS scores were seen 1 week after rehabilitation with surgical obturators at T2 (27.3 ±12.6) and at least 1 week after fabrication with definitive obturators at T4 (15.7 ±5.6). A comparison of the scores at T2, T3, and T4 found significant differences (P≤.001). Considerable improvement was seen in QOL with the fabrication of surgical obturators in participants who had received maxillectomy/debridement for mucormycosis. This improvement had a linear relationship with the time elapsed and the consecutive fitting of the obturators with healing.


Electrospinning of commercial guar-gum: Effects of purification and filtration.
Guar gums of two different commercial sources were successfully electrospun on both mica and copper tape at several concentrations starting from 1% (w/w). The electrospun fibers formed with the raw materials were not uniform and presented aggregates and beads within the fibers. Two different purification procedures and a filtration sequence with different pore size membranes were applied to enhance galactomannan solution homogeneity and solubility. The consequence was improved fiber morphology. We observed that the precipitation step, within the purification procedure, produced changes in the molar mass distribution and yielded different fiber diameter. Furthermore, spherical aggregates between fibers and within them disappeared after the sequential filtration. The resulting electrospun fiber diameter decreased with membrane pore diameter reduction. We conclude that the filtration process is responsible for molecular disentanglement, as well as disaggregation, which leads to improved electrospun galactomannan fiber morphology.


Use of the Basophil Activation Test May Reduce the Need for Drug Provocation in Amoxicillin-Clavulanic Allergy.
Reports of selective reactions to clavulanic acid (CLV) have increased in recent decades because of its increased prescription in combination with amoxicillin (AX) as AX-CLV. Basophil activation test (BAT) is used for diagnosing beta-lactam immediate hypersensitivity and is the only available in vitro assay for diagnosing patients with immediate hypersensitivity to CLV. However, few studies, and with limited numbers of patients have been published. The aim of this study was to establish the sensitivity, specificity, and negativization rates of BAT to AX and CLV. We studied 115 patients with immediate allergic reactions after AX-CLV treatment, 57 with selective reactions to AX (group A), 58 with selective reactions to CLV (group B), and 28 tolerant subjects. BAT was performed with AX in group A and with CLV in group B. A 4-year follow-up study was performed in patients with an initial positive BAT result. The overall sensitivity of BAT was 55%, specificity 89%, and positive predictive value (PPV) 96%. For group A, sensitivity was 47%, specificity 93%, and PPV 93%; for group B, sensitivity was 62%, specificity 89%, and PPV 92%. Follow-up study showed a faster negativization rate of BAT for group A, with around 40% of patients becoming negative at 12 months in both groups. The high PPV of BAT to CLV shows its potential value as a complementary tool to the allergological workup of patients with immediate allergic reactions after AX-CLV treatment. Importantly, the assay should be done within the first 12 months after the reaction to reduce false-negative results.


Abnormal zinc metabolism in type II diabetes mellitus.
Zinc metabolism in 20 patients with stable type II diabetes mellitus was investigated. Twenty-five percent of these patients had depressed serum zinc concentrations, and all demonstrated hyperzincuria. Urinary zinc loss was greater when proteinuria was present and correlated with the mean serum glucose concentration. Studies of gastrointestinal zinc absorption suggested zinc malabsorption in patients with type II diabetes mellitus. Glucose infusion in normal dogs produced hyperzincuria without a diminution in serum zinc. It is concluded that hyperzincuria, resulting from a glucose-mediated process that is not osmotic, interacts with impaired zinc absorption to produce zinc deficiency in patients with type II diabetes mellitus.


Comparative study of the usefulness of risk score assessment in the early stages of COVID-19 affected pregnancies: Omicron variant versus previous variants.
To evaluate the utility of the risk score in assessing the current status and prognosis of COVID-19 in pregnancy. Seventy-seven cases affected before the Omicron variant epidemic and 50 pregnant cases affected by the Omicron variant were included. The risk score consists of maternal background, current condition, and examination findings. We determined the risk score in the early stages of disease onset. There were no significant differences in the maternal or gestational ages between the groups. The risk score was significantly lower in the After-Group patients (those affected during the Omicron epoch), while 14.3% of the Before-Group patients (those affected during the pre-Delta and Delta epochs), experienced a worsening of disease after the visit to the center, whereas none of the After-Group patients did. The Before Group's frequency of risk score items was higher among the two groups for "fever for ≥48 h," "mild pneumonia image," and "blood tests," whereas "disease onset 14 days after the second vaccination" was increased in After Group. The blood test parameters for platelet count, C-reactive protein, and D-dimer levels were not significantly different between the groups. The risk score system appeared superior in detecting deteriorating cases. There were no cases of post-illness deterioration in the After-Group, suggesting that cases of the Omicron variant in pregnancy may have had a less severe course compared to that of previous variants. However, there was no significant difference between the groups in terms of a specific blood test evaluation, suggesting the need for a combined evaluation of cases affected during pregnancy.


Severity of nasal obstruction can predict the anxiety status of patients with allergic rhinitis but not patients with vasomotor rhinitis.
Allergic rhinitis (AR) is a chronic systemic allergic disease with an association reported between allergy and anxiety. The objective of the current study was therefore to investigate and retrospectively evaluate the factors that may possibly induce state and trait anxiety among patients with AR and vasomotor rhinitis (VMR). A total of 253 outpatients suffering from AR and 108 suffering from VMR were assessed for nasal symptoms experienced during the 14 days prior to assessment. All patients scored the severity of their symptoms on a visual analogue scale (VAS) and answered the State-Trait Anxiety Inventory (STAI) questionnaire to evaluate their state (STAI-S) and trait (STAI-T) anxiety scores. Pearson's correlation and multiple regression tests were employed to assess correlations between symptom scores and anxiety status scores. The STAI-S/T scores were not significantly different between AR and VMR patients; however, for AR patients, STAI-S and STAI-T were significantly associated with nasal obstruction and the total nasal symptom scores. Multiple regression analysis further demonstrated that only nasal obstruction significantly influenced both STAI-S and STAI-T scores. In contrast, nasal itching and total nasal symptom scores were significantly correlated with the STAI-S and STAI-T scores in VMR patients, whereas continuous sneezing VAS scores were significantly associated with only the STAI-S score. However, multiple regression analysis showed that the associations between any of the 4 nasal symptom scores and STAI-S and STAI-T scores were not significant. Nasal obstruction may play a prominent role in mediation of anxiety in patients suffering from AR, but not in patients suffering from VMR.


Upregulated miR-125b mitigates inflammation, astrocyte activation, and dysfunction of spinal cord injury by inactivating the MAPK pathway.
Since the abnormal expression of miR-125b in spinal cord injury (SCI) and the regulatory effect of miR-125b on the MAPK pathway have been expounded, we attempt to investigate whether miR-125b exerts a regulatory effect on SCI by modulating the MAPK pathway. A SCI rat model was established. The rats were treated with miR-125b antagomir or agomir, and their motor function affected by miR-125b was further detected by Basso-Beattie-Bresnahan (BBB) scoring. The histopathological changes and neuronal loss in the spinal cord were evaluated using hematoxylin-eosin and Nissl staining. Microglia-conditioned medium (MCM) was prepared and further used to treat the astrocytes, the activation of which was evaluated via immunofluorescence staining. The expressions of miR-125b, inflammation-related factors (IL-6, IL-1β, TNF-α, and IL-10), and MAPK pathway-related proteins (p38, ERK1/2, and JNK1/2 as well as their phosphorylated (p) forms) in the spinal cord, serum, and MCM-treated astrocytes of rats were determined by reverse-transcription quantitative polymerase chain reaction (RT-qPCR), enzyme-linked immunosorbent assay, and Western blot. MiR-125b was lowly expressed in SCI-modeled rats. MiR-125b inhibitor aggravated the impaired motor function, the disorder within the tissue, astrocyte activation, and neuron loss in the spinal cord tissues of SCI-modeled rats, while miR-125b mimic did oppositely. MiR-125b inhibitor enhanced the levels of IL-6, IL-1β, TNF-α, p38, p-p38, p-ERK1/2, and p-JNK1/2, whilst reducing that of IL-10. Contrarily, miR-125b mimic exerted the opposite effects in SCI-modeled rats and MCM-treated astrocytes. Up-regulation of miR-125b mitigates inflammation, astrocyte activation, and dysfunction in SCI by inactivating the MAPK pathway.


EPR image based oxygen movies for transient hypoxia.
Chronic hypoxia strongly affects the malignant state and resistance to therapy for tumors. Transient hypoxia has been hypothesized, but not proven to be more deleterious. Electron paramagnetic resonance imaging (EPRI) provides non-invasive, quantitative imaging of static pO₂ in vivo. Dynamic EPRI produces pO₂ movies, enabling non-invasive assessment of in vivo pO₂ changes, such as transient hypoxia. Recent developments have been made to enable Dynamic EPRI. Maximally spaced projection sequencing has been implemented to allow for more accurate and versatile acquisition of EPRI data when studying dynamic systems. Principal component analysis filtering has been employed to enhance SNR. Dynamic EPRI studies will provide temporally resolved oxygen movies necessary to perform in vivo studies of physiologically relevant pO₂ changes in tumors. These oxygen movies will allow for the localization/quantification of transient hypoxia and will therefore help to disentangle the relationship between chronic and transient hypoxia, in order to better understand their roles in therapeutic optimization and outcome.


Pembrolizumab versus chemotherapy for patients with esophageal squamous cell carcinoma enrolled in the randomized KEYNOTE-181 trial in Asia.
In the randomized phase III KEYNOTE-181 study, pembrolizumab prolonged overall survival (OS) compared with chemotherapy as second-line therapy in patients with advanced esophageal cancer and programmed death-ligand 1 (PD-L1) combined positive score (CPS) ≥10. We report a post hoc subgroup analysis of patients with esophageal squamous cell carcinoma (ESCC) enrolled in KEYNOTE-181 in Asia, including patients from the KEYNOTE-181 China extension study. Three hundred and forty Asian patients with advanced/metastatic ESCC were enrolled in KEYNOTE-181, including the China cohort. Patients were randomly assigned 1 : 1 to receive pembrolizumab 200 mg every 3 weeks for ≤2 years or investigator's choice of paclitaxel, docetaxel, or irinotecan. OS, progression-free survival, response, and safety were analyzed without formal comparisons. OS was evaluated based on PD-L1 CPS expression level. In Asian patients with ESCC, median OS was 10.0 months with pembrolizumab and 6.5 months with chemotherapy [hazard ratio (HR), 0.63; 95% CI 0.50-0.80; nominal P < 0.0001]. Median progression-free survival was 2.3 months with pembrolizumab and 3.1 months with chemotherapy (HR, 0.79; 95% CI 0.63-0.99; nominal P = 0.020). Objective response rate was 17.1% with pembrolizumab and 7.1% with chemotherapy; median duration of response was 10.5 months and 7.7 months, respectively. In patients with PD-L1 CPS <1 tumors (pembrolizumab versus chemotherapy), the HR was 0.99 (95% CI 0.56-1.72); the HR (95% CI) for death was better for patients with PD-L1 CPS cut-offs >1 [CPS ≥1, 0.57 (0.44-0.75); CPS ≥5, 0.56 (0.41-0.76); CPS ≥10, 0.53 (0.37-0.75)]. Treatment-related adverse events were reported in 71.8% of patients in the pembrolizumab group and 89.8% in the chemotherapy group; grade 3-5 events were reported in 20.0% and 44.6%, respectively. Pembrolizumab monotherapy demonstrated promising efficacy in Asian patients with ESCC, with fewer treatment-related adverse events than chemotherapy. PD-L1 CPS ≥1 is an appropriate cut-off and a predictive marker of pembrolizumab efficacy in Asian patients with ESCC.


EBV transformed B cell lines present to antigen specific T cell clones determinants different from those recognized by antibody in an HLA-DR linked restricted fashion.
Human B cells nonspecifically activated by Epstein Barr virus (EBV) can present tetanus toxoid (TT) antigen to TT specific helper T cell lines and T cell clones. Presentation of TT antigen by EBV-B cells did not require the presence of TT specific B cells and did not involve B cell surface immunoglobulins. Immunosorbent purified antibody to TT added in great excess to EBV-B cells pulsed with TT for 18 hr did not inhibit the capacity of the EBV-B cells to present TT antigen. Furthermore, EBV-B cells induced proliferation in T cells in the presence of urea denatured TT which contained less than 1% TT reactive with serum anti TT antibody. Studies of TT presentation by a panel of EBV-B cells obtained from HLA typed donors indicated that T cell recognition of TT presented by EBV-B cells was MHC restricted by products of the HLA-D region and some of which differed from serologically defined HLA-DR. These results indicate that the immunogenic moiety presented by EBV activated human B cells consists of antigenic determinants, which differ from those recognized by serum antibody, and of HLA-DR linked MHC determinants. This moiety is similar to that presented by monocytes. The implications of these findings for the amplification of the immune response by activated B cells are discussed.


Recovery of protein from green leaves: Overview of crucial steps for utilisation.
Plant leaves are a major potential source of novel food proteins. Till now, leaf protein extraction methods mainly focus on the extraction of soluble proteins, like rubisco protein, leaving more than half of all protein unextracted. Here, we report on the total protein extraction from sugar beet leaves (Beta vulgaris L.) by a traditional thermal extraction method consisting of mechanical pressing, heating to 50 °C and centrifugation. The resulting streams (i.e. supernatant, green-protein pellet and fibrous pulp) were characterised in terms of composition, physical structure and processing options. The protein distributed almost equally over the supernatant, pellet and pulp. This shows that thermal precipitation is an unselective process with respect to fractionation between soluble (rubisco) and insoluble (other) proteins. About 6% of the total protein could be extracted as pure rubisco (90% purity) from the supernatant. Surfactants commonly used for protein solubilisation could hardly re-dissolve the precipitated proteins in the pellet phase, which suggested that irreversible association was induced between the co-precipitated proteins and cell debris. Thus, the extraction of this protein will require prevention of their co-precipitation, and should take place in the original juice solution.


Analysis of driver injury severity in wrong-way driving crashes on controlled-access highways.
For more than five decades, wrong-way driving (WWD) has been notorious as a traffic safety issue for controlled-access highways. Numerous studies and efforts have tried to identify factors that contribute to WWD occurrences at these sites in order to delineate between WWD and non-WWD crashes. However, none of the studies investigate the effect of various confounding variables on the injury severity being sustained by the at-fault drivers in a WWD crash. This study tries to fill this gap in the existing literature by considering possible variables and taking into account the ordinal nature of injury severity using three different ordered-response models: ordered logit or proportional odds (PO), generalized ordered logit (GOL), and partial proportional odds (PPO) model. The findings of this study reveal that a set of variables, including driver's age, condition (i.e., intoxication), seatbelt use, time of day, airbag deployment, type of setting, surface condition, lighting condition, and type of crash, has a significant effect on the severity of a WWD crash. Additionally, a comparison was made between the three proposed methods. The results corroborate that the PPO model outperforms the other two models in terms of modeling injury severity using our database. Based on the findings, several countermeasures at the engineering, education, and enforcement levels are recommended.


Regulation of Airway Inflammation by G-protein Regulatory Motif Peptides of AGS3 protein.
Respiratory diseases such as asthma, chronic obstructive pulmonary disease (COPD), and lung infections have critical consequences on mortality and morbidity in humans. The aims of the present study were to examine the mechanisms by which CXCL12 affects MUC1 transcription and airway inflammation, which depend on activator of G-protein signaling (AGS) 3 and to identify specific molecules that suppress CXCL12-induced airway inflammation by acting on G-protein-coupled receptors. Herein, AGS3 suppresses CXCL12-mediated upregulation of MUC1 and TNFα by regulating Gαi. We found that the G-protein regulatory (GPR) motif peptide in AGS3 binds to Gαi and downregulates MUC1 expression; in contrast, this motif upregulates TNFα expression. Mutated GPR Q34A peptide increased the expression of MUC1 and TGFβ but decreased the expression of TNFα and IL-6. Moreover, CXCR4-induced dendritic extensions in 2D and 3D matrix cultures were inhibited by the GPR Q34A peptide compared with a wild-type GPR peptide. The GPR Q34A peptide also inhibited CXCL12-induced morphological changes and inflammatory cell infiltration in the mouse lung, and production of inflammatory cytokines in bronchoalveolar lavage (BAL) fluid and the lungs. Our data indicate that the GPR motif of AGS3 is critical for regulating MUC1/Muc1 expression and cytokine production in the inflammatory microenvironment.


Subversion of Endoscopic Breast Reconstruction Surgery: Nipple-sparing Mastectomy and Immediate Reconstruction with a Latissimus Dorsi Flap/Latissimus Dorsi Flap and Implant Through a Single Axillary Incision.
Our team has created a novel endoscopic technique (ET) to solve the problems of the former minimally invasive surgery (Inconvenient operation for ET, high cost for robotic technique, long-operation time for both) for breast cancer patients requiring nipple-sparing mastectomy and immediate breast reconstruction (BR) with a latissimus dorsi flap/latissimus dorsi flap and implant. This easy-to-grasp method will benefit more patients.


The impact of RASopathy-associated mutations on CNS development in mice and humans.
The RAS signaling pathway is involved in the regulation of developmental processes, including cell growth, proliferation, and differentiation, in the central nervous system (CNS). Germline mutations in the RAS signaling pathway genes are associated with a group of neurodevelopmental disorders, collectively called RASopathy, which includes neurofibromatosis type 1, Noonan syndrome, cardio-facio-cutaneous syndrome, and Costello syndrome. Most mutations associated with RASopathies increase the activity of the RAS-ERK signaling pathway, and therefore, most individuals with RASopathies share common phenotypes, such as a short stature, heart defects, facial abnormalities, and cognitive impairments, which are often accompanied by abnormal CNS development. Recent studies using mouse models of RASopathies demonstrated that particular mutations associated with each disorder disrupt CNS development in a mutation-specific manner. Here, we reviewed the recent literatures that investigated the developmental role of RASopathy-associated mutations using mutant mice, which provided insights into the specific contribution of RAS-ERK signaling molecules to CNS development and the subsequent impact on cognitive function in adult mice.


Development of a fluorescence polarization binding assay for asialoglycoprotein receptor.
Asialoglycoprotein receptor (ASGP-R) has been actively investigated for targeted delivery of therapeutic agents into hepatocytes because this receptor is selectively and highly expressed in liver and has a high internalization rate. Synthetic cluster glycopeptides (e.g., triGalNAc) bind with high affinity to ASGP-R and, when conjugated to a therapeutic agent, can drive receptor-mediated uptake in liver. We developed a novel fluorescent polarization (FP) ASGP-R binding assay to determine the binding affinities of ASGP-R-targeted molecules. The assay was performed in 96-well microplates using membrane preparations from rat liver as a source of ASGP-R and Cy5 fluorophore-labeled triGalNAc synthetic ligand as a tracer. This high-throughput homogeneous assay demonstrates advantages over existing multistep methods in that it minimizes both time and resources spent in determining binding affinities to ASGP-R. At the optimized conditions, a Z' factor of 0.73 was achieved in a 96-well format.


Taurine abated subacute dichlorvos toxicity.
The aim of the study was to determine the effects of taurine in rats exposed to subacute dichlorvos toxicity. Fifty rats were weighed and assigned into five groups of ten rats each. The groups received: distilled water, soya oil (1 ml/kg), taurine (50 mg/kg), dichlorvos (10 mg/kg) and the combination treatment group received taurine first and then dichlorvos 30 min later. The treatments were administered once daily by oral gavage for 4 weeks. The rats were euthanized and blood samples were collected after the termination of the study. Serum samples were analysed for malondialdehyde concentration and activities of antioxidant enzymes (superoxide dismutase and catalase). Dichlorvos increased malondialdehyde concentration and reduced the activities of superoxide dismutase and catalase. There was attenuation of malondialdehyde concentration and improvement of activities of superoxide dismutase (P = 0.0273) and catalase (P < 0.0001) in rats treated with taurine. It is postulated that taurine ameliorated dichlorvos-induced oxidative stress through the reduction of malondialdehyde concentration and the enhancement of activities of antioxidant enzymes.


The use of sialidase therapy for respiratory viral infections.
DAS181 is an inhaled bacterial sialidase which functions by removing sialic acid (Sia) from the surface of epithelial cells, preventing attachment and subsequent infection by respiratory viruses that utilize Sia as a receptor. DAS181 is typical of bacterial sialidases in cleaving Sia α2-3 and Sia α2-6 linkages, and it also has a demonstrated effect against acetylated and hydroxylated forms of Sia. The potency of the compound has been enhanced by coupling the active sialidase with an amphiregulin tag, allowing a longer duration of action and minimizing spread to the systemic circulation. DAS181 is now in Phase II development for the treatment of influenza, and it has also demonstrated activity in individual cases of parainfluenza in immunosuppressed patients. Continued evaluation of the roles and activities of bacterial sialidases is required to expand the range of successful antiviral therapies targeting Sia or its derivatives.


N-Formylmethionyl-leucyl-phenylalanine: Different releasing effects on human neutrophils and rat mast cells.
N-Formylmethionyl-leucyl-phenylalanine (FMLP) is a synthetic chemotactic peptide which induced beta-glucuronidase and lysozyme release from human neutrophils treated with cytochalasin B. FMLP-releasing effects were rapid and dose dependent. Unlike other secretagogues of neutrophils (e.g., zymosan and immune complexes), FMLP secretory activity was not modulated by acetylcholine, which by itself did not release lysosomal enzymes from human neutrophils. Isolated rat mast cells did not respond to FMLP, which has been demonstrated to release histamine from human neutrophils. Two markers of rat mast cell secretory granules, histamine and beta-glucuronidase, were assayed, but the results were negative for both. In the same experimental conditions, 48/80 released histamine and the enzyme: the ratio of the net percentage release of beta-glucuronidase to the net percentage release of histamine was congruent 0.4.


High prevalence of autoantibodies against the nuclear high mobility group (HMG) protein SSRP1 in sera from patients with systemic lupus erythematosus, but not other rheumatic diseases.
To evaluate the presence of autoantibodies to the high mobility group (HMG) structure specific recognition protein I (SSRP1) in sera from patients with systemic lupus erythematosus (SLE) or other rheumatic diseases. Antibodies to SSRP1(anti-SSRP1) were measured in sera from patients with SLE, Sjogren's syndrome (SS), ulcerative colitis (UC), systemic sclerosis (SSc), rheumatoid arthritis (RA), and sera from healthy individuals by both an enzyme-linked immunoassay (ELISA) and Western blotting (WB) using the recombinant SSRP1 N-terminus as antigen. We found 28.8% of the sera from patients with SLE contained anti-SSRPI by both ELISA and WB assay, compared to 8.3% of the sera from healthy individuals. When the 40 sera from patients with other autoimmune diseases were tested, only 2 sera (5%) from individuals with SS showed a moderate reactivity to SSRPI in both ELISA and WB assays. The results show that anti-SSRPI can be identified in sera from patients with SLE, but not with other rheumatic diseases and may thus help the diagnosis of SLE in the presence of appropriate clinical findings.


Substrate-modulated cytochrome P450 17A1 and cytochrome b5 interactions revealed by NMR.
The membrane heme protein cytochrome b5 (b5) can enhance, inhibit, or have no effect on cytochrome P450 (P450) catalysis, depending on the specific P450, substrate, and reaction conditions, but the structural basis remains unclear. Here the interactions between the soluble domain of microsomal b5 and the catalytic domain of the bifunctional steroidogenic cytochrome P450 17A1 (CYP17A1) were investigated. CYP17A1 performs both steroid hydroxylation, which is unaffected by b5, and an androgen-forming lyase reaction that is facilitated 10-fold by b5. NMR chemical shift mapping of b5 titrations with CYP17A1 indicates that the interaction occurs in an intermediate exchange regime and identifies charged surface residues involved in the protein/protein interface. The role of these residues is confirmed by disruption of the complex upon mutagenesis of either the anionic b5 residues (Glu-48 or Glu-49) or the corresponding cationic CYP17A1 residues (Arg-347, Arg-358, or Arg-449). Cytochrome b5 binding to CYP17A1 is also mutually exclusive with binding of NADPH-cytochrome P450 reductase. To probe the differential effects of b5 on the two CYP17A1-mediated reactions and, thus, communication between the superficial b5 binding site and the buried CYP17A1 active site, CYP17A1/b5 complex formation was characterized with either hydroxylase or lyase substrates bound to CYP17A1. Significantly, the CYP17A1/b5 interaction is stronger when the hydroxylase substrate pregnenolone is present in the CYP17A1 active site than when the lyase substrate 17α-hydroxypregnenolone is in the active site. These findings form the basis for a clearer understanding of this important interaction by directly measuring the reversible binding of the two proteins, providing evidence of communication between the CYP17A1 active site and the superficial proximal b5 binding site.


Kinetic Chemiluminescence as a Method for Oxidative Stress Evaluation in Examinations of Patients with Type 2 Diabetes Mellitus.
We propose a new approach to evaluation of oxidative stress based on kinetic chemiluminescence: measurement of antioxidant and pro-oxidant activities of the plasma. The study included 50 patients with type 2 diabetes mellitus receiving peroral hypoglycemic therapy. In addition to the above parameters, the levels of TBA-reactive products, inflammation markers, clotting parameters, and biochemical values were studied. The new method provides information on oxidative stress in patients with type 2 diabetes mellitus irrespective of the clinical and laboratory values. The use of this method in complex with the clinical, laboratory, and instrumental studies allows comprehensive evaluation of patient's status for the diagnosis and choice of therapy.


7-Meth-oxy-penta-cyclo-[5.4.0.02,6.03,10.05,9]undec-ane-8,11-dione.
The structure of 7-meth-oxy-penta-cyclo-[5.4.0.02,6.03,10.05,9]undecane-8,11-dione, C12H12O3, at 150 K has monoclinic (P21/c) symmetry. The penta-cyclo-undecane cage compound is composed of four five-membered rings, a planar four-membered ring and a six-membered ring in a boat conformation fused into a closed strained-cage framework. All of the five-membered rings adopt an envelope conformation.


Cardiovascular risk factors among people with drug-resistant tuberculosis in Uganda.
Tuberculosis (TB) and its risk factors are independently associated with cardiovascular disease (CVD). We determined the prevalence and associations of CVD risk factors among people with drug-resistant tuberculosis (DRTB) in Uganda. In this cross-sectional study, we enrolled people with microbiologically confirmed DRTB at four treatment sites in Uganda between July to December 2021. The studied CVD risk factors were any history of cigarette smoking, diabetes mellitus (DM) hypertension, high body mass index (BMI), central obesity and dyslipidaemia. We used modified Poisson regression models with robust standard errors to determine factors independently associated with each of dyslipidaemia, hypertension, and central obesity. Among 212 participants, 118 (55.7%) had HIV. Overall, 196 (92.5%, 95% confidence interval (CI) 88.0-95.3) had ≥ 1 CVD risk factor. The prevalence; 95% CI of individual CVD risk factors was: dyslipidaemia (62.5%; 55.4-69.1), hypertension (40.6%; 33.8-47.9), central obesity (39.3%; 32.9-46.1), smoking (36.3%; 30.1-43.1), high BMI (8.0%; 5.0-12.8) and DM (6.5%; 3.7-11.1). Dyslipidaemia was associated with an increase in glycated haemoglobin (adjusted prevalence ratio (aPR) 1.14, 95%CI 1.06-1.22). Hypertension was associated with rural residence (aPR 1.89, 95% CI 1.14-3.14) and previous history of smoking (aPR 0.46, 95% CI 0.21-0.98). Central obesity was associated with increasing age (aPR 1.02, 95%CI 1.00-1.03), and elevated diastolic blood pressure (aPR 1.03 95%CI 1.00-1.06). There is a high prevalence of CVD risk factors among people with DRTB in Uganda, of which dyslipidaemia is the commonest. We recommend integrated services for identification and management of CVD risk factors in DRTB.


Functionalized Multi-Wall Carbon Nanotubes Enhance Transfection and Expression Efficiency of Plasmid DNA in Fish Cells.
DNA vaccines are considered to be the most promising method against infectious diseases in the aquaculture industry. In the present study, we investigated the potency of ammonium group-functionalized multi-walled carbon nanotubes (MWCNTs) in enhancing the transfection and expression efficiency of plasmid DNA (pEGFP-vp5) in Ctenopharyngodon idellus kidney (CIK) cells. Agarose gel shift assay results show that ammonium group-functionalized carbon nanotubes are able to condense DNA in varying degrees. Scanning electron microscope (SEM) images shows that CIK cells show a great affinity for MWCNTs-NH₃⁺ and the CNTs covering the cell surface tend to orient their tips perpendicularly to the cell surface, and appear to be "needle-pricking the cells". Transmission electron microscope (TEM) images confirmed that MWCNTs-NH₃⁺ penetrate the cell membranes and are widely dispersed in the CIK cell. Real-time PCR was used to detect the transfection efficiency through the expression of the outer capsid protein (VP5). The results showed that the MWCNTs-NH₃⁺:DNA complexes are able to transfect CIK cells effectively at different charge ratio than naked DNA. Subsequent studies confirmed that both functional groups and charge ratio are important factors that determine the transfection efficiency of plasmid DNA. All these results indicated that MWCNTs-NH₃⁺:DNA complexes could be suitable for developing DNA vaccine for the control of virus infection in the aquaculture industry.


Venous thromboembolism prevention in intracerebral hemorrhage: A systematic review and network meta-analysis.
To summarize and compare the effectiveness of pharmacological thromboprophylaxis to pneumatic compression devices (PCD) for the prevention of venous thromboembolism in patients with acute intracerebral hemorrhage. MEDLINE, PUBMED, EMBASE, and CENTRAL were systematically searched to identify randomized and non-randomized studies that compared each intervention directly to each other or against a common control (hydration, anti-platelet agents, stockings) in adults with acute spontaneous intracerebral hemorrhage. Two investigators independently screened the studies, extracted data, and appraised risk of bias. Studies with a high risk of bias were excluded from our final analysis. The primary outcome was the occurrence of venous thromboembolism (proximal deep vein thrombosis or pulmonary embolism) in the first 30 days. 8,739 articles were screened; four articles, all randomized control trials, met eligibility criteria. Bayesian network meta-analysis was performed to calculate risk estimates using both fixed and random effects analyses. 607 patients were included in the network analysis. PCD were associated with a significant decrease in venous thromboembolism compared to control (OR: 0.43, 95% Credible Limits [CrI]: 0.23-0.80). We did not find evidence of statistically significant differences between pharmacological thromboprophylaxis and control (OR: 0.93, 95% CrI: 0.19-4.37) or between PCD and pharmacological thromboprophylaxis (OR: 0.47, 95% CrI: 0.09-2.54). PCDs are superior to control interventions, but meaningful comparisons with pharmacotherapy are not possible due to a lack of data. This requires further exploration via large pragmatic clinical trials. PROSPERO: CRD42018090960.


Biomechanical analysis of the knee with partial anterior cruciate ligament disruption: quantitative evaluation using an electromagnetic measurement system.
To investigate the biomechanical function of anterior cruciate ligament (ACL) remnants in ACL-deficient knees with both partial and complete tears. Twenty partial ACL-deficient (group P), 20 complete ACL-deficient (group C), and 40 contralateral ACL-intact knees were examined. The end point during the Lachman test, side-to-side differences of KT-1000 measurements, and the pivot shift test were evaluated. Additionally, the side-to-side difference of anterior tibial translation during the Lachman test and the acceleration during the pivot shift test were calculated using an electromagnetic measurement system (EMS). The end point was found in 9 patients in group P, whereas it was not detected in group C. In KT-1000 measurements, the mean side-to-side differences were 3.8 ± 2.4 mm in group P and 5.4 ± 2.3 mm in group C. There was a significant difference between these 2 groups (P < .05). In the pivot shift test evaluation in group P, one patient was evaluated as grade 0, 17 patients as grade 1+, and 2 patients as grade 2+. In group C, 10 patients were evaluated as grade 1+, 9 patients as grade 2+, and one patient as grade 3+. Using the EMS, mean side-to-side differences during the Lachman test were 3.1 ± 2.1 mm in group P and 7.2 ± 3.2 mm in group C. The anterior-posterior displacement in group P was significantly less than that in group C (P < .05). In the quantitative pivot shift test, the mean acceleration in the contralateral ACL-intact knees was -632.7 ± 254.5 mm/s(2), whereas it was -1107.5 ± 398.9 mm/s(2) in group P and -1652.2 ± 754.9 mm/s(2) in group C. Significant differences were detected between the 3 knee conditions (P < .05). The quantitative assessments of knees with partial ACL ruptures during the Lachman test and the pivot shift test using the EMS showed less laxity than did knees with complete ACL tears, whereas their laxity was greater than the contralateral knees with intact ACLs. Level III, diagnostic study of nonconsecutive patients.


Transcriptomic and proteomic data provide new insights into cold-treated potato tubers with T- and D-type cytoplasm.
Tuber-omics in potato with the T- and D-types of cytoplasm showed different sets of differentially expressed genes and proteins in response to cold storage. For the first time, we report differences in gene and protein expression in potato (Solanum tuberosum L.) tubers possessing the T- or D-type cytoplasm. Two F1 diploid reciprocal populations, referred to as T and D, were used. The pooling strategy was applied for detection of differentially expressed genes (DEGs) and differentially expressed proteins (DEPs) in tubers consisting of extreme chip colour after cold storage. RNA and protein bulks were constructed from contrasting phenotypes. We recognized 48 and 15 DEGs for the T and D progenies, respectively. DEPs were identified in the amyloplast and mitochondrial fractions. In the T-type cytoplasm, only 2 amyloplast-associated and 5 mitochondria-associated DEPs were detected. Of 37 mitochondria-associated DEPs in the D-type cytoplasm, there were 36 downregulated DEPs in the dark chip colour bulks. These findings suggest that T- and D-type of cytoplasm might influence sugar accumulation in cold-stored potato tubers in different ways. We showed that the mt/nucDNA ratio was higher in D-possessing tubers after cold storage than in T progeny. For the D-type cytoplasm, the pt/nucDNA ratio was higher for tubers characterized by dark chip colour than for those with light chip colour. Our findings suggest that T- and D-type cytoplasm might influence sugar accumulation in cold-stored potato tubers in different ways.


Effects of 2-Year Physical Activity and Dietary Intervention on Adrenarchal and Pubertal Development: The PANIC Study.
Childhood overweight has been linked to earlier development of adrenarche and puberty, but it remains unknown if lifestyle interventions influence sexual maturation in general populations. To investigate if a 2-year lifestyle intervention influences circulating androgen concentrations and sexual maturation in a general population of children. We conducted a 2-year physical activity and dietary intervention study in which 421 prepubertal and mostly normal-weight 6- to 9-year-old children were allocated either to a lifestyle intervention group (119 girls, 132 boys) or a control group (84 girls, 86 boys). The main outcome measures were serum dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulfate (DHEAS), androstenedione (A4), and testosterone concentrations, and clinical adrenarchal and pubertal signs. The intervention and control groups had no differences in body size and composition, clinical signs of androgen action, and serum androgens at baseline. The intervention attenuated the increase of DHEA (P = .032), DHEAS (P = .001), A4 (P = .003), and testosterone (P = .007) and delayed pubarche (P = .038) in boys but it only attenuated the increase of DHEA (P = .013) and DHEAS (P = .003) in girls. These effects of lifestyle intervention on androgens and the development of pubarche were independent of changes in body size and composition, but the effects of intervention on androgens were partly explained by changes in fasting serum insulin. A combined physical activity and dietary intervention attenuates the increase of serum androgen concentrations and sexual maturation in a general population of prepubertal and mostly normal-weight children, independently of changes in body size and composition.


Bioequivalence and food effect assessment for vildagliptin/metformin fixed-dose combination tablets relative to free combination of vildagliptin and metformin in Japanese healthy subjects.
To assess the bioequivalence of vildagliptin/metformin fixeddose combination (FDC) tablets (50/250 mg and 50/500 mg) to free combinations of vildagliptin and metformin and the effect of food on the pharmacokinetics (PK) of vildagliptin and metformin following administration of 50/500 mg FDC tablets. Two openlabel, randomized, single-center, singledose, 2-period crossover studies were conducted in Japanese healthy male volunteers. Participants were administered vildagliptin/ metformin FDC tablets (study I: 50/250 mg, study II: 50/500 mg) or their free combinations under fasted condition. Food effect (standard Japanese breakfast: fat, 20 - 30% with ~ 600 kcal in total) was assessed during an additional period in study II (50/500 mg). PK parameters (AUC, C(max), t(max), t(1/2)) were calculated for vildagliptin and metformin. In both studies, vildagliptin/metformin FDC tablets were bioequivalent to their respective free combinations. Administration of FDC tablets after meals had no effect on vildagliptin PK parameters. The rate of absorption of metformin decreased when administered under fed condition, as reflected by a prolonged t(max) (3 hours in fasted state vs. 4 hours in fed state) and decrease in C(max) by 26%, however, the extent of absorption (AUC(last)) was similar to that in the fasted state. Vildagliptin/metformin FDC tablets were bioequivalent to their free combinations. Food decreased the C(max) of metformin by 26%, while AUC(last) was unchanged, consistent with previous reports. No food effect was observed on the C(max) or AUC(last) of vildagliptin. Thus, food had no clinically relevant effects on the PK of metformin or vildagliptin.


Disruption of extracellular redox balance drives persistent lung fibrosis and impairs fibrosis resolution.
Lung fibrosis is a devastating outcome of various diffuse parenchymal lung diseases. Despite rigorous research efforts, the mechanisms that propagate its progressive and nonresolving nature remain enigmatic. Oxidative stress has been implicated in the pathogenesis of lung fibrosis. However, the role of extracellular redox state in disease progression and resolution remains largely unexplored. Here, we show that compartmentalized control over extracellular reactive oxygen species (ROS) by aerosolized delivery of recombinant extracellular superoxide dismutase (ECSOD) suppresses an established bleomycin-induced fibrotic process in mice. Further analysis of publicly available microarray, RNA-seq and single-cell RNAseq datasets reveals a significant decrease in ECSOD expression in fibrotic lung tissues that can be spontaneously restored during fibrosis resolution. Therefore, we investigate the effect of siRNA-mediated ECSOD depletion during the established fibrotic phase on the self-limiting nature of the bleomycin mouse model. Our results demonstrate that in vivo knockdown of ECSOD in mouse fibrotic lungs impairs fibrosis resolution. Mechanistically, we demonstrate that transforming growth factor (TGF)-β1 downregulates endogenous ECSOD expression, leading to the accumulation of extracellular superoxide via Smad-mediated signaling and the activation of additional stores of latent TGF-β1. In addition, depletion of endogenous ECSOD during the fibrotic phase in the bleomycin model induces an apoptosis-resistant phenotype in lung fibroblasts through unrestricted Akt signaling. Taken together, our data strongly support the critical role of extracellular redox state in fibrosis persistence and resolution. Based on these findings, we propose that compartment-specific control over extracellular ROS may be a potential therapeutic strategy for managing fibrotic lung disorders.


Lost to Care and Back Again: Patient and Navigator Perspectives on HIV Care Re-engagement.
Engagement in HIV care is critical to achieve viral suppression and ultimately improve health outcomes for people living with HIV (PLWH). However, maintaining their engagement in care is often a challenging goal. Utilizing patient navigators, trained in an adapted ARTAS intervention, to help re-engage out-of-care PLWH has proven to be a valuable resource. This qualitative study describes the encounters between PLWH (n = 11) and their care re-engagement navigators (n = 9). Participants were interviewed in-person; interviews were transcribed and analyzed using the strengths model of case management. PLWH shared how working with navigators increased their motivation to return to HIV care and assisted them to overcome barriers that were a hindrance to care engagement. Navigators described a strengths-based approach to working with their clients, thus helping facilitate PLWH care re-engagement goals and successes. Results from this study may inform the development of effective HIV navigation programs to re-engage out-of-care PLWH, often the hardest-to-engage.


Specific alteration of the form of selenium in fetal liver on maternal methylmercury treatment.
We have recently reported that maternal administration of methylmercury caused a striking increase in the selenium concentration in fetal liver accompanied by a decrease in selenium-dependent glutathione peroxidase (GSH-Px) activity. These changes resulted in the lowered bioavailability of selenium as far as the GSH-Px activity was concerned. The present study demonstrated that maternal administration of methylmercury caused a specific alteration of the form of selenium in fetal liver. Sephadex G-200 gel filtration of liver cytosols revealed an additional major peak of selenium in the fetal livers of mice treated with methylmercury. This peak was not present in the liver, kidney, or placenta of mothers treated with methylmercury.


SparkEC: speeding up alignment-based DNA error correction tools.
In recent years, huge improvements have been made in the context of sequencing genomic data under what is called Next Generation Sequencing (NGS). However, the DNA reads generated by current NGS platforms are not free of errors, which can affect the quality of downstream analysis. Although error correction can be performed as a preprocessing step to overcome this issue, it usually requires long computational times to analyze those large datasets generated nowadays through NGS. Therefore, new software capable of scaling out on a cluster of nodes with high performance is of great importance. In this paper, we present SparkEC, a parallel tool capable of fixing those errors produced during the sequencing process. For this purpose, the algorithms proposed by the CloudEC tool, which is already proved to perform accurate corrections, have been analyzed and optimized to improve their performance by relying on the Apache Spark framework together with the introduction of other enhancements such as the usage of memory-efficient data structures and the avoidance of any input preprocessing. The experimental results have shown significant improvements in the computational times of SparkEC when compared to CloudEC for all the representative datasets and scenarios under evaluation, providing an average and maximum speedups of 4.9[Formula: see text] and 11.9[Formula: see text], respectively, over its counterpart. As error correction can take excessive computational time, SparkEC provides a scalable solution for correcting large datasets. Due to its distributed implementation, SparkEC speed can increase with respect to the number of nodes in a cluster. Furthermore, the software is freely available under GPLv3 license and is compatible with different operating systems (Linux, Windows and macOS).


Developmental programming: Prenatal testosterone-induced changes in epigenetic modulators and gene expression in metabolic tissues of female sheep.
Prenatal testosterone (T)-treated female sheep manifest peripheral insulin resistance and tissue-specific changes in insulin sensitivity with liver and muscle manifesting insulin resistance accompanied by inflammatory, oxidative and lipotoxic state. In contrast, visceral (VAT) and subcutaneous (SAT) adipose tissues are insulin sensitive in spite of VAT manifesting changes in inflammatory and oxidative state. We hypothesized that prenatal T-induced changes in tissue-specific insulin resistance arise from disrupted lipid storage and metabolism gene expression driven by changes in DNA and histone modifying enzymes. Changes in gene expression were assessed in liver, muscle and 4 adipose (VAT, SAT, epicardiac [ECAT] and perirenal [PRAT]) depots collected from control and prenatal T-treated female sheep. Prenatal T-treatment increased lipid droplet and metabolism genes PPARA and PLIN1 in liver, SREBF and PLIN1 in muscle and showed a trend for decrease in PLIN2 in PRAT. Among epigenetic modifying enzymes, prenatal T-treatment increased expression of 1) DNMT1 in liver and DNMT3A in VAT, PRAT, muscle and liver; 2) HDAC1 in ECAT, HDAC2 in muscle with decrease in HDAC3 in VAT; 3) EP300 in VAT and ECAT; and 4) KDM1A in VAT with increases in liver histone acetylation. Increased lipid storage and metabolism genes in liver and muscle are consistent with lipotoxicity in these tissues with increased histone acetylation likely contributing to increased liver PPARA. These findings are suggestive that metabolic defects in prenatal T-treated sheep may arise from changes in key genes mediated, in part, by tissue-specific changes in epigenetic-modifying enzymes.