Food specific skin-test reactivity in atopic subjects.
It has been proposed that a permanent or transitory increase in gut permeability is an important facet in the development of food allergy. If this occurs, then individuals with a history of a specific food allergy should have a higher incidence of immunological reactivity to other food allergens as compared to food tolerant subjects. To test this hypothesis, we evaluated the prevalence of food-specific IgE responses by skin-prick testing in 60 individuals. Subjects were classified by a history of food allergy (shrimp hypersensitivity) and atopic status. Prevalence of skin-prick test reactivity to shrimp and a panel of nine other food extracts was determined. Skin-test reactivity to shrimp was related both a history of shrimp sensitivity and atopic status. However, the prevalence of skin-test reactivity to other foods was not related to clinical history of shrimp allergy, although it was related to atopy. A subset of subjects with shrimp allergy had multiple positive skin reactions to many of the nine other foods. This reactivity was associated with a history of pulmonary symptoms following shrimp ingestion. In general our results do not support non-specific increases in gut permeability being important in the development of food allergy.


Effect of Ion Irradiation on Nanoindentation Fracture and Deformation in Silicon Carbide.
Silicon carbide is desirable for many nuclear applications, making it necessary to understand how it deforms after irradiation. Ion implantation combined with nanoindentation is commonly used to measure radiation-induced changes to mechanical properties; hardness and modulus can be calculated from load-displacement curves, and fracture toughness can be estimated from surface crack lengths. Further insight into indentation deformation and fracture is required to understand the observed changes to mechanical properties caused by irradiation. This paper investigates indentation deformation using high-resolution electron backscatter diffraction (HR-EBSD) and Raman spectroscopy. Significant differences exist after irradiation: fracture is suppressed by swelling-induced compressive residual stresses, and the plastically deformed region extends further from the indentation. During focused ion beam cross-sectioning, indentation cracks grow, and residual stresses are modified. The results clarify the mechanisms responsible for the modification of apparent hardness and apparent indentation toughness values caused by the compressive residual stresses in ion-implanted specimens. The online version contains supplementary material available at 10.1007/s11837-021-04636-8.


Knowledge gaps and uncertainties about epilepsy: findings from an ethnographic study in China.
Epilepsy represents one of the major brain disorders worldwide. In China, research into how much people with epilepsy know about their condition appears limited. Drawing on data collected as part of a large ethnographic study, we present the experiences and views of Chinese people with epilepsy and their family members, to identify knowledge gaps and uncertainties about epilepsy within selected urban and rural communities. We also examine how respondents' demographic characteristics influence their knowledge, understanding, and beliefs about epilepsy. We found knowledge and understanding of epilepsy to be uneven and context specific. Hereditary factors were most frequently cited as a potential cause, although their impact remained unclear. Western medicalization of epilepsy appears less evident in the reports of rural informants, where traditional beliefs continue to shape definitions and treatment. Societal differences within these communities set boundaries on knowledge acquisition. Plotted against these differences, we suggest strategies for proposed educational/psychosocial intervention programs.


The care of elderly patients by elderly physicians.
To examine the content of care that elderly patients receive from physicians greater than or equal to 65 years of age, data from the 1981 National Ambulatory Medical Care Survey were analyzed. Compared to physicians aged 35 to 54 years, elderly physicians devoted a larger proportion of their practices to the care of elderly patients. In caring for these patients, elderly physicians spent more time per visit than did younger physicians and were more likely to conduct general examinations and provide counseling. In contrast, elderly physicians were less likely to perform mental status or vision examinations. Elderly physicians were also less likely to provide telephone follow-up for those patients greater than or equal to 75 years of age and more likely to discharge patients greater than or equal to 65 years of age without any specific follow-up planned. These data suggest that the content of care for elderly patients may differ substantially depending upon the physician's age.


A challenging, unpredictable world for people with autism spectrum disorder.
Autism is a pervasive neurodevelopmental disorder characterized by impairment of communication and social interaction, as well as by high levels of repetitive and ritualistic behaviours. This last dimension results in major difficulties in daily life: clinical reports of individuals with Autism Spectrum Disorder (ASD) show that they present tantrums as a response to change, or restricted interests and repetitive behaviours in order to prevent or minimize change. Such a crucial need to maintain sameness suggests substantial differences in how the ASD brain predicts the environment, and this might have a fundamental role in the deficit revealed in the highly unpredictable social world. Several lines of evidence indicating difficulties in generating or using predictions in ASD due to atypical information processing will be presented in this review. For instance, several studies have revealed that people with ASD demonstrate a unique profile of cognitive abilities, with strategies that depend to an abnormally large extent on sensory systems, at the expense of more integrative processing requiring an awareness of contextual subtleties necessary for prediction. At a more elementary level, patients with autism manifest unusual processing of unpredictable events, which might be rooted in a basic difference in how the brain orients to changing, novel sensory stimuli. This review presents results from ERPs and fMRI studies illustrating the psychophysiological mechanisms and neural bases underlying such phenomena in ASD. We propose that such dysfunction in the ability to build flexible prediction in ASD may originate from impaired top-down influence over a variety of sensory and higher level information processing, a physiopathological hypothesis which dovetails with the cortical under connectivity current theory.


Effects of resveratrol on inflammation and oxidative stress induced by the uremic toxin indoxyl sulfate in Murine macrophage-like RAW 264.7.
Indoxyl sulfate (IS) is a uremic toxin produced by the gut microbiota that commonly accumulates in patients with chronic kidney disease (CKD) and can be harmful. Resveratrol is a polyphenol with properties that attenuate oxidative stress and inflammation. This study aims to evaluate the effect of resveratrol against the damage caused by IS in RAW 264.7 murine macrophages. Cells were treated with 0, 250, 500 and 1000 μmol/L of IS, in the presence of 50 μmol/L of resveratrol. The mRNA and protein expressions of erythroid-related nuclear factor 2 (Nrf2) and nuclear factor kappa-B (NF-κB) were measured using rt-PCR and Western blot analysis, respectively. Malondialdehyde (MDA) and reactive oxygen species (ROS) levels were also analyzed. As a result, it was demonstrated that resveratrol induces the activation of the Nrf2 pathway that enhances cytoprotective response. IS upregulated the NF-κB expression and downregulated the Nrf2 expression. In contrast, resveratrol treatment significantly reduced the MDA and ROS production and inhibited the IS-induced expression of NF-κB in macrophage-like RAW 264.7. In conclusion, resveratrol can mitigate inflammation and oxidative stress caused by uremic toxins produced by the gut microbiota, such as IS.


PAF stimulates cAMP formation in P388D1 macrophage-like cells via the formation and secretion of prostaglandin E2 in an autocrine fashion.
The role of cAMP in the formation of prostaglandin E2 (PGE2) was investigated in bacterial lipopolysaccharide (LPS)-primed P388D1 macrophage-like cells stimulated with platelet activating factor (PAF). cAMP levels and PGE2 secretion were correlated with stimulation by PAF or ionomycin. Indomethacin inhibited cAMP formation induced by PAF, but not PGE2-stimulated cAMP production. Inositol (1,4,5)-trisphosphate levels were strongly reduced by exogenous PGE2 and increased by H-89, an inhibitor of PKA. However, exogenous PGE2 did not affect PAF-stimulated PGE2 formation. These results suggest that cAMP levels in P388D1 cells are regulated by PGE2 in an autocrine fashion. Evidence is presented that this feedback mechanism regulates inositol (1,4,5)-triphosphate levels in these cells, while PGE2 formation is not affected.


Correlation between promoter methylation of p14(ARF), TMS1/ASC, and DAPK, and p53 mutation with prognosis in cholangiocarcinoma.
To study the methylation status of genes that play a role in the p53-Bax mitochondrial apoptosis pathway and its clinical significance in cholangiocarcinoma. Out of 36 cases cholangiocarcinoma patients from April 2000 to May 2005 were collected.Promoter hypermethylation of DAPK, p14(ARF), and ASC were detected by methylation-specific PCR on cholangiocarcinoma and normal adjacent tissues samples. Mutation of the p53 gene was examined by automated sequencing. Correlation between methylation of these genes and/or p53 mutation status with clinical characteristics of patients was investigated by statistical analysis. We found 66.7% of 36 cholangiocarcinoma patients had methylation of at least one of the tumor suppressor genes analyzed. p53 gene mutation was found in 22 of 36 patients (61.1%). Combined p53 mutation and DAPK, p14(ARF), and/or ASC methylation was detected in 14 cases (38.9%). There were statistically significant differences in the extent of pathologic biology, differentiation, and invasion between patients with combined p53 mutation and DAPK, p14(ARF), and/or ASC methylation compared to those without (P < 0.05). The survival rate of patients with combined DAPK, p14(ARF), and ASC methylation and p53 mutation was poorer than other patients (P < 0.05). Our study indicates that methylation of DAPK, p14(ARF), and ASC in cholangiocarcinoma is a common event. Furthermore, p53 mutation combined with DAPK, p14(ARF), and/or ASC methylation correlates with malignancy and poor prognosis.


[Significance of hereditary predisposition factors in the development and course of pulmonary tuberculosis].
Comprehensive clinical, genetic, epidemiological, and immunogenetic studies have established that tuberculosis is a multifactorial disease whose development and natural history are due to the interaction of environmental factors, primarily, to infection and hereditary predisposition. Associations between tuberculosis and various HLA loci B antigens, as well as HLA locus D-DR2 antigen have been detected in different populations. The HLA genotype was shown to differ in patients with favourable and unfavourable tuberculosis natural history. The comprehensive study of genetic markers allows one to make a prompt assessment of the nature of a tuberculous process, to define its prognosis and to correct treatment.


[Relationship of the activity of cardiac cholinergic mechanisms to the state of the sympathetic innervation and its noradrenaline content in hypoxia].
Preliminary sympathectomia depletes acetylcholine (ACh) in the heart of rabbits under hypoxia. In these conditions the inhibitory action of ACh on the rat isolated heart is reduced under the noradrenaline content fall, while under increase it is potentiated. Under hypoxia noradrenaline increases concentration of potassium in the myocardium, thus stimulating ACh formation and activity. It is suggested that under deep hypoxia suppression of the sympathetic mechanisms causes functional isolation of the heart from nervous effects.


BMI and pelvimetry help to predict the duration of laparoscopic resection for low and middle rectal cancer.
In rectal cancer surgery, recent studies have found associations between clinical factors, especially pelvic parameters, and surgical difficulty; however, their findings are inconsistent because the studies use different criteria. This study aimed to evaluate common clinical factors that influence the operative time for the laparoscopic anterior resection of low and middle rectal cancer. Patients who underwent laparoscopic radical resection of low and middle rectal cancer from January 2018 to December 2020 were retrospectively analyzed and classified according to the operative time. Preoperative clinical and magnetic resonance imaging (MRI)-related parameters were collected. Logistic regression analysis was used to identify factors for predicting the operative time. In total, 214 patients with a mean age of 60.3 ± 8.9 years were divided into two groups: the long operative time group (n = 105) and the short operative time group (n = 109). Univariate analysis revealed that the male sex, a higher body mass index (BMI, ≥ 24.0 kg/m2), preoperative treatment, a smaller pelvic inlet (< 11.0 cm), a deeper pelvic depth (≥ 10.7 cm) and a shorter intertuberous distance (< 10.1 cm) were significantly correlated with a longer operative time (P < 0.05). However, only BMI (OR 1.893, 95% CI 1.064-3.367, P = 0.030) and pelvic inlet (OR 0.439, 95% CI 0.240-0.804, P = 0.008) were independent predictors of operative time. Moreover, the rate of anastomotic leakage was higher in the long operative time group (P < 0.05). Laparoscopic rectal resection is expected to take longer to perform in patients with a higher BMI or smaller pelvic inlet.


Regulation of replication termination by Reb1 protein-mediated action at a distance.
DNA transactions driven by long-range protein-mediated inter- and intrachromosomal interactions have been reported to influence gene expression. Here, we report that site-specific replication termination in Schizosaccharomyces pombe is modulated by protein-mediated interactions between pairs of Ter sites located either on the same or on different chromosomes. The dimeric Reb1 protein catalyzes termination and mediates interaction between Ter sites. The Reb1-dependent interactions between two antiparallel Ter sites in cis caused looping out of the intervening DNA in vitro and enhancement of fork arrest in vivo. A Ter site on chromosome 2 interacted pairwise with two Ter sites located on chromosome 1 by chromosome kissing. Mutational inactivation of the major interacting Ter site on chromosome 1 significantly reduced fork arrest at the Ter site on chromosome 2, thereby revealing a cooperative mechanism of control of replication termination.


Are probiotics detectable in human feces after oral uptake by healthy volunteers?
Assessment of the presence of probiotic bacteria in feces after oral ingestion. Probiotic bacteria are said to have beneficial effects on the host. As a precondition for any effect, probiotic strains must survive passage through the gastrointestinal tract. The feces of seven volunteers were analyzed for the presence of probiotic strains after one week's oral ingestion of each of six commercially available products: E. coli Nissle 0.5-5 x 10(9) cells (Mutaflor), Enterococcus faecium SF 68 7.5 x 10(7) cells (Bioflorin), Lactobacillus acidophilus and Bifidobacterium infantis both 1 x 10(9) cells (Infloran), Lactobacillus gasseri and Bifidobacterium longum both 1 x 10(8) cells (Omniflora), Lactobacillus casei rhamnosus 1 x 10(9) cells (Antibiophilus), and yoghurt enriched with Lactobacillus casei Immunitas 1 x 10(10) cells (Actimel). Ten colonies were selected from each stool sample, and DNA was extracted and typed using random amplification of polymorphic DNA (RAPD). Typing patterns of the ingested probiotics and the fecal isolates were compared. Fingerprints identical to the ingested probiotic strains were recovered from fecal samples of 4/7 volunteers after one week of Mutaflor, from 4/6 after taking Bioflorin, and from 1/6 after Infloran. Cultivation of strains of the same species from fecal specimens was negative after consumption of Antibiophilus, Omniflora and Actimel. After oral consumption of probiotics, E. coli and enterococci could be detected in stool samples (57% and 67%, respectively). In contrast, with only one exception, ingested lactobacilli and bifidobacteria could not be detected in human feces.


A multiplex blood-based assay targeting DNA methylation in PBMCs enables early detection of breast cancer.
The immune system can monitor tumor development, and DNA methylation is involved in the body's immune response to tumors. In this work, we investigate whether DNA methylation alterations in peripheral blood mononuclear cells (PBMCs) could be used as markers for early detection of breast cancer (BC) from the perspective of tumor immune alterations. We identify four BC-specific methylation markers by combining Infinium 850 K BeadChips, pyrosequencing and targeted bisulfite sequencing. Based on the four methylation markers in PBMCs of BC, we develop an efficient and convenient multiplex methylation-specific quantitative PCR assay for the detection of BC and validate its diagnostic performance in a multicenter cohort. This assay was able to distinguish early-stage BC patients from normal controls, with an AUC of 0.940, sensitivity of 93.2%, and specificity of 90.4%. More importantly, this assay outperformed existing clinical diagnostic methods, especially in the detection of early-stage and minimal tumors.


Consecutive entosis stages in human substrate-dependent cultured cells.
Entosis, or cell death by invading another cell, is typical for tumor epithelial cells. The formation of cell-in-cell structures is extensively studied in suspension cultures, but remains poorly understood in substrate-dependent cells. Here, we used electron, confocal and time-lapse microscopy in combination with pharmacological inhibition of intracellular components to study the kinetics of entosis using two human substrate-dependent tumor cultures, A431 and MCF7. In total, we identified and characterized five consecutive stages of entosis, which were common for both examined cell lines. We further demonstrated that actin filaments in the entotic as well as invading cells were crucial for entosis. Microtubules and the Golgi apparatus of entotic cells provided membrane expansion required for internalization of the invading cell. Depolymerization of microfilaments and microtubules, and disintegration of the Golgi complex inhibited entosis. We confirmed the presence of adhesive junctions and discovered the formation of desmosomes between the invading and entotic cells. The internalized cell was shown to be degraded due to the lysosomal activation in both cells whereas the disintegration of the Golgi apparatus did not affect the process. Thus, in the substrate-dependent cultures, entosis requires microfilaments, microtubules and the Golgi complex for cell invasion, but not for internalized cell degradation.


Chemokines and the tissue-specific migration of lymphocytes.
Tissue-selective trafficking of memory and effector T and B lymphocytes is mediated by unique combinations of adhesion molecules and chemokines. The discovery of several related epithelial-expressed chemokines (TECK/CCL25 in small intestine, CTACK/CCL27 in skin, and MEC/CCL28 in diverse mucosal sites) now highlights an important role for epithelial cells in controlling homeostatic lymphocyte trafficking, including the localization of cutaneous and intestinal memory T cells, and of IgA plasma cells. Constitutively expressed epithelial chemokines may help determine the character of local immune responses and contribute to the systemic organization of the immune system.


Intracelluar delivery of A20 protein inhibits TNFα-induced NF-κB activation.
A20 (also known as TNFAIP3) is a potent anti-inflammatory protein that suppresses many intracellular signaling pathways induced by inflammatory cytokines and bacterial and viral pathogens. The anti-inflammatory function of A20 depends on its modulation of or binding to polyubiquitin chains on key signaling proteins in the nuclear factor-κB (NF-κB) pathway. To test whether A20 can be used as therapeutic agent in these inflammatory diseases, we prepared a recombinant cell-penetrating form of A20 (TAT-A20) for intracellular delivery and examined its effect on tumor necrosis factor-α (TNFα)-induced NF-κB activation. We observed that TAT-A20 was effectively transduced into cells within 30 min, whereas A20 protein without TAT motive was not. TAT-A20 also inhibited NF-κB induction in fibroblasts stimulated with TNFα. These results suggest that increasing intracellular level of A20 can be an effective means to suppress NF-κB activation and treat inflammatory diseases.


Effects of IGF-I, rGH, FGF, EGF and NCS on DNA-synthesis, cell proliferation and morphology of chondrocytes isolated from rat rib growth cartilage.
The effects of IGF-I, rGH, FGF, EGF and NCS on DNA-synthesis were analyzed in resting, proliferative and hypertrophic chondrocytes obtained by fractionation. Proliferation and morphology were studied on non-fractionated cells. The highest stimulation of DNA-synthesis was induced by NCS followed by IGF-I at all stages of chondrocyte differentiation. DNA-synthesis was also stimulated by a low concentration of FGF (1 microgram/1) in proliferative and hypertrophic chondrocytes, while FGF in a higher concentration (10 micrograms/1) had no significant mitogenic effect. Cell proliferation was stimulated by both NCS and IGF-I, whereas FGF and EGF only caused morphological changes. Our data indicate that IGF-I is the main serum growth factor regulating growth and proliferation by interacting with chondrocytes at all stages of differentiation.


Anaerobic Digestion of Cattle Manure Influenced by Swirling Jet Induced Hydrodynamic Cavitation.
In this work, a modified swirling jet-induced cavitation has been employed for increasing anaerobic digestion efficiency of cattle manure. The hydrodynamic cavitation (HC) treatment improved the organic matter solubilization and the anaerobic biodegradability of cattle manure. The degree of disintegration increased by 5.8, 8.9, and 15.8% after the HC treatment at 6.0, 7.0, and 8.0 bars, respectively. However, the HC treatment at 7.0 bars had better results in terms of methane production. This result may be attributed to the possible formation of toxic and refractory compounds at higher inlet pressures, which could inhibit the methanization process. Further, total Kjeldahl nitrogen content was found to decrease with increasing inlet pressures, as the pH and the turbulent mixing favored the ammonia stripping processes. HC treatment decreased the viscosity of the treated cattle manure, favoring the manure pumping and mixing. Considerations on the energy input due to the HC pre-treatment and the energy output due to the enhanced methane yield have been presented. A positive energy balance can be obtained looking at the improved operational practices in the anaerobic digesters after the implementation of the HC pre-treatment.


Relationship between estimated and observed heparin sensitivity indices in cardiac and thoracic aortic surgery.
Little evidence exists on the relationship between the estimated heparin sensitivity index (HSI) based on commonly available clinical and laboratory data and observed HSI in the adult population. This retrospective study assessed the relationship between the observed and estimated HSIs. This study was conducted in an academic, single-institution setting. Patients aged ≥ 20 years who underwent cardiac and thoracic aortic surgery and requiring cardiopulmonary bypass were included. Clinical and laboratory data, including age, sex, and platelet count, were collected. The fibrinogen-albumin ratio index was calculated by dividing the fibrinogen value by the albumin value, multiplied by 10.The HSI was calculated using the formula: (activated clotting time after initial heparin administration-baseline activated clotting time)/initial heparin dose (IU/kg). The estimated HSI was based on the results of multiple regression analysis that included clinically relevant factors. The intraclass correlation coefficient between the observed and estimated HSIs was used to assess. In total, 560 patients with valid activated clotting time (ACT) values after initial heparin administration were included in the final analysis to explore associated factors using the estimated HSI. Multiple regression analysis revealed that hemodialysis, platelet count, fibrinogen-to-albumin ratio index, baseline activated clotting time, and initial heparin dose were significantly associated with the HSI. The mean (standard deviation) observed and estimated HSIs were 1.38 (0.43) and 1.55 (0.13), respectively, with an intra-class correlation coefficient of 0.10. The correlation between the observed and estimated HSIs was low, and a formula with high accuracy for estimating the HSI is needed.


Comparison of the osteogenic effectiveness of an autogenous demineralised dentin matrix and Bio-Oss® in bone augmentation: a systematic review and meta-analysis.
This study aimed to evaluate the clinical efficacy and histological outcomes of autogenous demineralised dentin matrix (ADDM) as bone graft material compared with Bio-Oss® in bone augmentation for the treatment of patients with oral bone deficits. Eight databases (PubMed, EMBASE, Cochrane Library, Science Direct, Scopus, Web of Science, CNKI, and WFPD) were searched for randomised controlled trials (RCT) performed from the date of inception of each database to July 2021. The Cochrane Collaboration's risk assessment tool was used to conduct the methodological quality assessment. Stata 15.0 software was used to perform data analysis. Seven RCTs including 220 patients were considered eligible for this study. No significant difference was found in the percentage of new bone formation (NBF) and implant stability quotient (ISQ). Patients who received ADDM grafting showed a significantly lower sinus height (SH) and percentage of residual graft material (RGM) compared with Bio-Oss® grafting. ADDM is as effective as Bio-Oss® in bone augmentation for oral bone defects.


Assessment of neuromuscular blockade produced by atracurium in frog sartorius muscle.
The neuromuscular effects of atracurium in frog sartorius muscle were studied and the results were compared with those obtained by tubocurarine in the same preparation. Single twitch (at 1 per 5 s) and tetanic stimulation (at 1-100 per s) of the motor nerve, stimulated with 5-10 V (maximum) and 0.2 ms pulse duration, were used to assess the neuromuscular blockade produced by atracurium and tubocurarine in the frog sartorius muscle. The results showed that atracurium was twice as potent as tubocurarine in reducing the amplitude of the indirectly-elicited twitch contractions in the frog sartorius muscle. The mean IC50 values (concentration to produce 50% inhibition of twitch tension) of atracurium and tubocurarine-induced depression of the indirectly-elicited twitch tension were 0.64 +/- 0.1 microM and 1.2 +/- 0.2 microM respectively (means +/- s.e., n = 6, P less than 0.001). Furthermore, atracurium had a shorter time of onset, time to 50% block and time to 95% recovery than tubocurarine.


Ocular pathologic findings in neonatal adrenoleukodystrophy.
Ocular pathology in a case of neonatal adrenoleukodystrophy (ALD) showed marked degeneration of photoreceptor cells throughout the retina including the macula. Macroscopically visible white opacities in the cortical vitreous of the posterior fundus were shown immunohistochemically and ultrastructurally to be macrophages containing bileaflet inclusions, a finding previously unreported. Additional findings included extensive loss of the retinal nerve fiber and ganglion cell layers, optic nerve degeneration, and bileaflet inclusions in retinal pigment epithelial cells and macrophages of the retina and optic nerve. Focal retinal pigmentary changes were observed that histologically were different from retinitis pigmentosa.


Invention and mystery in hypotrich DNA.
Hypotrichs have evolved extraordinary ways of organizing, manipulating, and replicating the DNA in their micronuclear and macronuclear genomes. Short macronuclear DNA molecules containing single genes are created by excision from chromosomes, accompanied by massive elimination of the germline DNA sequences between genes. Germline genes themselves are interrupted by multiple noncoding segments called internal eliminated segments, or IESs, that divide genes into multiple macronuclear-destined segments, or MDSs. The functional significance of this organization is unknown. Over evolutionary time IESs accumulate mutations rapidly, are inserted into or excised from genes, and shift position along DNA molecules. MDSs are ligated to create functional genes when IESs are spliced out of micronuclear DNA during macronuclear development. MDSs in some germline genes are in scrambled disorder and become unscrambled in association with IES elimination. Replication of DNA in the macronucleus is accomplished by organization of replication enzymes and factors into a structure that sweeps through the macronucleus to replicate the many millions of gene-sized DNA molecules. The significance of many of the bizarre DNA phenomena in the evolutionary/functional success of hypotrichs is still unclear.


Risk factors and precipitants of long-term disability in community mobility: a cohort study of older persons.
Relatively little is known about why older persons develop long-term disability in community mobility. To identify the risk factors and precipitants for long-term disability in walking a quarter mile and driving a car. Prospective cohort study from March 1998 to December 2009. Greater New Haven, Connecticut. 641 persons, aged 70 years or older, who were active drivers or nondisabled in walking a quarter mile. Persons who were physically frail were oversampled. Candidate risk factors were assessed every 18 months. Disability in community mobility and exposure to potential precipitants, including illnesses or injuries leading to hospitalization or restricted activity, were assessed every month. Disability that lasted 6 or more consecutive months was considered long-term. 318 (56.0%) and 269 (53.1%) participants developed long-term disability in walking and driving, respectively. Seven risk factors were independently associated with walking disability and 8 were associated with driving disability; the strongest associations for each outcome were found for older age and lower score on the Short Physical Performance Battery. The precipitants had a large effect on long-term disability, with multivariate hazard ratios for each outcome greater than 6.2 for hospitalization and greater than 2.4 for restricted activity. The largest differences in absolute risk were generally observed in participants with a specific risk factor who were subsequently hospitalized. The observed associations may not be causal. The severity of precipitants was not assessed. The effect of the precipitants may have been underestimated because their exposure after the initial onset of disability was not evaluated. Long-term disability in community mobility is common among older persons. Multiple risk factors, together with subsequent precipitants, greatly increase the likelihood of long-term mobility disability. National Institute on Aging, National Institutes of Health.


Edaravone Attenuates the Proinflammatory Response in Amyloid-β-Treated Microglia by Inhibiting NLRP3 Inflammasome-Mediated IL-1β Secretion.
Microglial activation is an important pathological feature in the brains of patients with Alzheimer's disease (AD), and amyloid-β (Aβ) peptides play a crucial role in microglial activation. In addition, edaravone (EDA) was recently shown to suppress oxidative stress and proinflammatory cytokine production in APPswePS1dE9 (APP/PS1) mice. However, the mechanism by which EDA inhibits the Aβ-induced proinflammatory response in microglia is poorly understood. The mitochondrial membrane potential (∆ψm) was evaluated using JC-1 staining. Intracellular reactive oxygen species (ROS) and mitochondrial ROS levels were detected using CM-H2DCFDA and MitoSOXTM Red, respectively. The levels of CD11b, NLRP3, pro-caspase-1 and manganese superoxide dismutase (SOD-2) were observed by western blotting, and the levels of interleukin-1beta (IL-1β) in culture supernatants were quantified using an ELISA kit. Aβ induced microglia activation and mitochondrial dysfunction. In addition, mitochondrial dysfunction was associated with ROS accumulation and activation of the NLRP3 inflammasome. Importantly, Aβ induced activation of the NLRP3 inflammasome, leading to caspase-1 activation and IL-1β release in microglia. Moreover, EDA obviously attenuated the depolarization of ∆ψm, reduced mitochondria-derived ROS production and increased SOD-2 activity, resulting in the suppression of NLRP3 inflammasome-mediated IL-1β secretion in Aβ-treated microglia. EDA is a mitochondria-targeted antioxidant and exhibits anti-inflammatory effects on Aβ-treated microglia.


Prior exposure to the carrier regulates rat immune responses to a conjugate vaccine.
Small vaccine antigens including peptides generally need to be linked to larger molecules or carriers in order to induce high levels of immune responses. The potential of unwanted immune responses to the carriers represents a major drawback for the conjugated vaccines. The carriers could also regulate the immune responses to the haptens and these effects need to be prevented in order to achieve adequate responses to the vaccines. We examined means to reduce the unwanted reactions to the carrier. For this purpose, we investigated whether prior exposure of rats to a human IgG(1) (hIgG(1)) carrier would affect their subsequent responses to an OVA peptide (i.e., OVA(173-196)). Prior exposure to the hIgG(1) carrier did not affect the T cell responses to the peptide antigen. However, IgG(1) Ab responses to the peptide antigen were enhanced while IgE Abs were reduced. These results show that responses to the hapten are not systematically relevant to the carrier pre-immunization and that the conjugate could achieve desired responses by selective immune responses suppression. Such models of vaccines with enhanced anti-hapten responses and reduced levels of potentially harmful responses could be of great interest for the development of new immune therapies.


Diagnosis and treatment of vaginal intraepithelial neoplasia.
To audit our experience of the diagnosis and treatment of vaginal intraepithelial neoplasia (VAIN) in Kandang Kerbau Women's Hospital, Singapore. The clinical records of the patients diagnosed as having VAIN at our institution in the calendar year 2009 were periodically reviewed until March 2011. There were 21 cases of VAIN. The mean follow-up duration was 18.2 months. The lesion grades were VAIN 1 in 9 patients (42.9%), VAIN 2 in 9 patients (42.9%), and VAIN 3 in 3 patients (14.3%). The mean patient age was 39.1 years. Sixteen patients (76.2%) were referred to our institution because of abnormal results to cytology tests and 4 patients (21.9%) were referred because of vaginal warts. The remaining patient was diagnosed from a surgical specimen. The diagnosis involved a cytology test, a colposcopic examination, an acetowhite test, a Schiller test, and a colposcopy-directed biopsy. The treatments varied and included watchful waiting but carbon dioxide laser vaporization was used the most frequently. After 6 months of follow-up 18 patients were considered cured, for an overall cure rate of 85.7%. Two patients wanted no treatment and disease remained persistent in 1 patient despite laser vaporization and intravaginal applications of imiquimod cream.


Protein kinase C: A potential therapeutic target for endothelial dysfunction in diabetes.
Protein kinase C (PKC) is a family of serine/threonine protein kinases that play an important role in many organs and systems and whose activation contributes significantly to endothelial dysfunction in diabetes. The increase in diacylglycerol (DAG) under high glucose conditions mediates PKC activation and synthesis, which stimulates oxidative stress and inflammation, resulting in impaired endothelial cell function. This article reviews the contribution of PKC to the development of diabetes-related endothelial dysfunction and summarizes the drugs that inhibit PKC activation, with the aim of exploring therapeutic modalities that may alleviate endothelial dysfunction in diabetic patients.


Efficacy of the 7-chloro-4-(3-hydroxy-benzilidenehydrazo)quinoline derivative against infection caused by Leishmania amazonensis.
The drugs currently available for leishmaniasis treatment have major limitations. In vitro and in vivo studies were performed to evaluate the effect of a quinoline derivative, Hydraqui (7-chloro-4-(3-hydroxy-benzilidenehydrazo)quinoline, against Leishmania amazonensis. In silico analyses of absorption, distribution, metabolism, excretion, and toxicity (ADMET) parameters were performed. Hydraqui showed significant in vitro anti-amastigote activity. Also, Hydraqui-treated mice exhibited high efficacy in lesion size (48.3%) and parasitic load (93.8%) reduction, did not cause hepatic and renal toxicity, and showed appropriate ADMET properties. Hydraqui presents a set of satisfactory criteria for its application as an antileishmanial agent.


Pharmacoinformatics-based investigation of bioactive compounds of Rasam (South Indian recipe) against human cancer.
Spice-rich recipes are referred to as "functional foods" because they include a variety of bioactive chemicals that have health-promoting properties, in addition to their nutritional value. Using pharmacoinformatics-based analysis, we explored the relevance of bioactive chemicals found in Rasam (a South Indian cuisine) against oxidative stress-induced human malignancies. The Rasam is composed of twelve main ingredients, each of which contains a variety of bioactive chemicals. Sixty-six bioactive compounds were found from these ingredients, and their structures were downloaded from Pubchem. To find the right target via graph theoretical analysis (mitogen-activated protein kinase 6 (MAPK6)) and decipher their signaling route, a network was built. Sixty-six bioactive compounds were used for in silico molecular docking study against MAPK6 and compared with known MAPK6 inhibitor drug (PD-173955). The top four compounds were chosen for further study based on their docking scores and binding energies. In silico analysis predicted ADMET and physicochemical properties of the selected compounds and were used to assess their drug-likeness. Molecular dynamics (MD) simulation modelling methodology was also used to analyse the effectiveness and safety profile of selected bioactive chemicals based on the docking score, as well as to assess the stability of the MAPK6-ligand complex. Surprisingly, the discovered docking scores against MAPK6 revealed that the selected bioactive chemicals exhibit varying binding ability ranges between - 3.5 and - 10.6 kcal mol-1. MD simulation validated the stability of four chemicals at the MAPK6 binding pockets, including Assafoetidinol A (ASA), Naringin (NAR), Rutin (RUT), and Tomatine (TOM). According to the results obtained, fifty of the sixty-six compounds showed higher binding energy (- 6.1 to - 10.6 kcal mol-1), and four of these compounds may be used as lead compounds to protect cells against oxidative stress-induced human malignancies.


Repeated surveillance of lead poisoning among children.
In 1980, a survey of lead poisoning was carried out among 592 children aged 1 to 9 years living within 900 m of a lead smelter in Santo Amaro, Brazil. From 1980 to 1985, the lead smelter carried out a number of major improvements aimed at reducing environmental pollution. In January, 1985, a second survey was carried out among a sample of 250 children living in this same geographical area. The geometric mean of zinc protoporphyrin (ZPP) concentration in whole blood was 1.17 (standard deviation = 1.5) mumol l-1. Blood lead concentrations (PbB) determined in a subsample of 53 children had an arithmetic mean and standard deviation of 1.77 +/- 1.00 mumol l-1, with 22 children showing PbB greater than or equal to 1.68 mumol l-1. Twenty-nine (11.6%) of the 250 children lived in houses where the lead content of the soil was greater than 10,000 ppm, and they presented higher ZPP levels than the rest of the population. Children with the habit of pica for soil had elevated ZPP levels. Comparing the results from the 1980 and 1985 surveys, slight improvements in ZPP and PbB levels were observed. However, new cases of lead poisoning are still occurring in the area. The soil is highly contaminated by lead and represents a long-lasting risk factor for child intoxication.


Comparative Analysis of the Interaction of the Primary Quinone QA in Intact and Mn-Depleted Photosystem II Membranes Using Light-Induced ATR-FTIR Spectroscopy.
In photosystem II, redox potential Em of primary quinone QA shifts by approximately +150 mV upon inactivation of the Mn4CaO5 cluster. Although this phenomenon is important in photoprotection, the molecular mechanism of the positive Em(QA-/QA) shift remains unclear. Here, we investigated the effect of Mn depletion on the interaction of QA using light-induced ATR-FTIR spectroscopy. It was shown that Mn depletion hardly changed a QA-/QA FTIR difference spectrum, indicating that the H-bond interaction of QA and its immediate surroundings were virtually unchanged by Mn4CaO5 cluster inactivation. On the basis of this result, a possible mechanism of the Em(QA-/QA) shift is discussed.


Superstructures of Organic-Polyoxometalate Co-crystals as Precursors for Hydrogen Evolution Electrocatalysts.
Molybdenum-based carbides and nitrides have been considered as catalysts for the hydrogen evolution reaction (HER). One of the challenges in using Mo-based HER electrocatalysts is establishing well-defined precursors which can be transformed into Mo-based carbides/nitrides with controllable structure and porosity. We report the synthesis of a series of superstructures consisting of organic-polyoxometalate co-crystals (O-POCs) as a new type of metal-organic precursor to synthesize Mo-based carbides/nitrides in a controlled fashion and to use them for efficient catalytic hydrogen production. This protocol enables to create electrocatalysts composed of abundant nanocrystallites and heterojunctions with tunable micro- and nanostructure and mesoporosity. The best performing electrocatalyst shows high HER activity and stability with a low overpotential of 162 mV at 100 mA cm-2 (in comparison to Pt/C with 263 mV), which makes it one of the best non-noble metal HER catalysts in alkaline media and seawater.


Method optimization for the determination of four mercury species by micro-liquid chromatography-inductively coupled plasma mass spectrometry coupling in environmental water samples.
A method based on the coupling microHPLC-microneb-ICPMS has been developed for Hg(II), MeHg+, EtHg+ and PhHg+ species. Gradient elution using methanol and l-cysteine at pH 3.0 allowed the chromatographic separation of all species in less than 13 min (total analysis time 15 min). The direct coupling of microLC to ICPMS through a Micromist nebulizer permits the analysis of environmental water without sample pretreatment and derivatization steps. Nebulizer type, organic modifier and column length were the main parameters tested. The methanol content and pH of the mobile phase greatly affected the retention time and sensitivity of the method. Key factors to obtain high signal to noise ratio, at concentrations below 1 microg L(-1), were found to be the nebulization step and traces of Hg present in the complexing agent. A detailed optimization of carrier and make up gas flow rates have enabled the nebulization of the methanol gradient elution with good mass transport efficiency, low organic solvent loading into the plasma and excellent precision. The performance of the microHPLC-microneb-ICPMS method developed was evaluated on a surface water sample filtered (0.22 microm) and spiked with 0.5 microg L(-1) (as Hg) of each species. Precision (R.S.D., n=6) for all species of Hg varied from 0.5 to 2.1%. Detection limit, defined as three times the standard deviation (n=6), ranged from 8 ng L(-1) for EtHg+ to 32 ng L(-1) for PhHg+ and was noticeably lower than those reported in previous LC-based methods. Accuracy was suitable with recoveries ranging from 85 to 100% when tested at two levels (0.5 and 10 microg L(-1)) in groundwater samples. Recovery was matrix affected when water samples of high salinity (depurated wastewater and seawater) were used.


The role of shear stress on cutaneous microvascular endothelial function in humans.
Previous studies suggest that exercise and heat stress improve cutaneous endothelial function, caused by increases in shear stress. However, as vasodilatation in the skin is primarily a thermogenic phenomenon, we investigated if shear stress alone without increases in skin temperature that occur with exercise and heat stress increases endothelial function. We examined the hypothesis that repeated bouts of brief occlusion would improve cutaneous endothelial function via shear stress-dependent mechanisms. Eleven males underwent a shear stress intervention (forearm occlusion 5 s rest 10 s) for 30 min, five times·week-1 for 6 weeks on one arm, the other was an untreated control. Skin blood flow was measured using laser-Doppler flowmetry, and endothelial function was assessed with and without NOS-inhibition with L-NAME in response to three levels of local heating (39, 42, and 44 °C), ACh administration, and reactive hyperaemia. Data are cutaneous vascular conductance (CVC, laser-Doppler/blood pressure). There were no changes in the control arm (all d ≤ 0.2, p > 0.05). In the experimental arm, CVC to 39 °C was increased after 3 and 6 weeks (d = 0.6; p ≤ 0.01). Nitric oxide contribution was increased after 6 weeks compared to baseline (d = 0.85, p < 0.001). Following skin heating to 42 °C and 44 °C, CVC was not different at weeks 3 or 6 (d ≤ 0.8, p > 0.05). For both 42 and 44 °C, nitric oxide contribution was increased after weeks 3 and 6 (d ≥ 0.4, p < 0.03). Peak and area-under-the-curve responses to ACh increased following 6 weeks (p < 0.001). Episodic increases in shear stress, without changes in skin or core temperature, elicit an increase in cutaneous microvascular reactivity and endothelial function.


Analysis of biomechanical effect of stem-end design in revision TKA using Digital Korean model.
The stem-end pain after revision total knee arthroplasty has been clinically reported around the diaphyseal region of the tibia, and the stem-end design has been recognized as one of the causes of the stem-end pain. But there are few reports about the biomechanical characteristics of different stem-end designs on revision total knee arthroplasty. The objective of this study is to investigate the biomechanical effect of the stem-end design in revision total knee arthroplasty. Three-dimensional finite element models of a tibia and implants for revision total knee arthroplasty with various stem-end design parameters were developed. The tibia model was chosen from 50 male cadavers to represent the standard Korean male. The contact pressures and von Mises stresses around the stem-end were calculated under axial loading. The longer stem length, the bigger stem diameter, the stronger press-fit, and no slot shape increased both peak contact pressures and von Mises stresses. The location of peak contact pressures coincided with the location of stem-end pain reported in clinical studies. Different stem-end designs altered the contact pressures or stress concentrations to the tip of the stem. The location of peak contact pressures or von Mises stresses suggested there may be a biomechanical factor to transfer load or generate stress concentration at the location of the stem-end pain in the diaphysis.


Arthroscopic Anterior Cruciate Ligament Primary Repair With Synthetic Augmentation and Fixation With the Knotless Suture Anchor.
An anterior cruciate ligament (ACL) tear is one of the most common ligament injuries in athletes. The arthroscopic ACL reconstruction procedure is the gold standard for treatment. However, the improvement in injury classification and suture materials has subsequently made arthroscopic ACL primary repair an alternative surgical treatment option. This Technical Note describes an arthroscopic ACL primary repair with synthetic augmentation made of several high-strength sutures and fixation with the knotless suture anchor. The reinforced synthetic material acts as a structural tie to support the ACL while it heals.


Relatively short term test re-test reliability of Neck Disability Index by long term test re-retest reliability method of Oswestry Disability Index in healthy office workers.
Assessment of local condition specific outcome measures in combination to obtain an idea about the disability status of the whole spine is a conventional method. Oswestry Disability Index (ODI) and Neck Disability Index (NDI) are two outcome measures used together. Test re-test reliability of ODI in healthy subjects has clinical importance and the test re-test reliability of NDI may have clinical significance. The purpose of this study is to investigate the test re-test reliability of the NDI by using long-term test re-test reliability method of ODI on healthy office workers. Participants who have no chronic neck pain history were included in the study. Subjects were assessed by the Turkish-NDI (e-forms) on 1st, 2nd, 4th, 8th, 15th, 30th days. 49 (20 female, 29 male) of 106 participants (57 female, 49 male) completed the study. Kolmogorov-Smirnov and Friedman tests were used. The difference between median score of each day (χ= 9.275, p > 0.05) was neither statistically nor clinically significant. NDI has test re-test reliability in healthy subjects over a 1-month time interval and the test re-test reliability is also valid in cases where both questionnaires are used in combination in this time interval.


Concentrations of nitrogen compounds are related to severe rhinovirus infection in infants. A time-series analysis from the reference area of a pediatric university hospital in Barcelona.
There is scarce information focused on the effect of weather conditions and air pollution on specific acute viral respiratory infections, such as rhinovirus (RV), with a wide clinical spectrum of severity. The aim of this study was to analyze the association between episodes of severe respiratory tract infection by RV and air pollutant concentrations (NOx and SO2 ) in the reference area of a pediatric university hospital. An analysis of temporal series of daily values of NOx and SO2 , weather variables, circulating pollen and mold spores, and daily number of admissions in the pediatric intensive care unit (PICU) with severe respiratory RV infection (RVi) in children between 6 months and 18 years was performed. Lagged variables for 0-5 days were considered. The study spanned from 2010 to 2018. Patients with comorbidities were excluded. One hundred and fifty patients were admitted to the PICU. Median age was 19 months old (interquartile range [IQR]: 11-47). No relationship between RV-PICU admissions and temperature, relative humidity, cumulative rainfall, or wind speed was found. Several logistic regression models with one pollutant and two pollutants were constructed but the best model was that which included average daily NOx concentrations. Average daily NOx concentrations were related with the presence of PICU admissions 3 days later (odds ratio per IQR-unit increase: 1.64, 95% confidence interval: 1.20-2.25)). This study has shown a positive correlation between NOx concentrations at Lag 3 and children's PICU admissions with severe RV respiratory infection. Air pollutant data should be taken into consideration when we try to understand the severity of RVis.


Ancestry informative markers derived from discriminant analysis of principal components provide important insights into the composition of crossbred cattle.
The cost of SNP genotyping to screen different breeds and to estimate the exact proportion of ancestry level is quite high, which can be compensated through deriving a small panel of ancestry informative markers (AIMs). Hence, we carried out the present study to provide an insight into ancestry level inferred from a panel of informative markers in the crossbred Vrindavani population developed at ICAR-IVRI, India. We have performed a new method i.e., discriminant analysis of principal components (DAPC) for the first time on the dataset of Vrindavani cattle. To confirm our method, we had performed DAPC on two other well-known crossbred cattle, i.e., Frieswal and Beefmaster. Three sets of panels (500, 1000 and 2000 markers) were tested for clustering of individuals. Among all the panels, we found the panel (1000 markers) with DAPC based contribution method was of the smallest size and comparatively of the highest accuracy.


Peroneal flap: How to harvest and clinical appraisal for head and neck reconstruction.
A peroneal flap, the boneless version of fibula flap, is considered as the equivalent of radial forearm flap of the lower leg. Because it is thinner than an anterolateral thigh (ALT) flap, the use of a peroneal flap is a viable option for the repair of soft tissue defects when a thin flap could bring about better functional and cosmetic outcomes. In this article, we describe the details of peroneal flap harvest and present our experience with the use of peroneal flaps for head and neck reconstruction. Between 1996 and 2017, a total of 265 peroneal flaps were used to reconstruct a variety of head and neck defects. With the same vascular anatomy and slight modifications to the harvesting technique of a fibula flap, a peroneal flap can be harvested within 1-2 h. All medical records were retrospectively reviewed. A peroneal flap can be harvested as different types of chimeric flaps to fit a variety of head and neck defects. The peroneal flap failure rate was 3.4% and the postoperative complication rate was 12.8%. A peroneal flap might be an alternative option for the reconstruction of head and neck defects.


Analysis of 15 cases from a monocentric cohort of 307 liver abscesses.
Out of the context of haematological patients, Candida sp. is rarely retrieved from pyogenic liver abscesses (PLA). Our objective was to assess the risk factors for occurrence, and clinical, microbiological characteristics, management and outcome of Candida pyogenic liver abscesses (C-PLA). We retrospectively analysed C-PLA cases and compared them to pyogenic liver abscesses exclusively due to bacteria (B-PLA) included in our monocentric database on liver abscesses. Unfavourable course was defined as the occurrence of a primary treatment failure (PTF), recurrence after an initial cure, or death within 3 months after diagnosis. Between 2010 and 2018, 15 C-PLA and 292 B-PLA were included. All C-PLA had a biliary origin and were polymicrobial. All patients with C-PLA had at least one comorbidity at risk for Candida infection and 7 (53.3%) presented with sepsis requiring an admission in intensive care unit. Median duration of antifungal treatment was 42 days [24-55]. In multivariate analysis, compared with B-PLA, a medical history of malignancy (OR 4.16; 95%CI 1.15-18.72) or liver abscess (OR 7.39; 95%CI 2.10-26.62), and sepsis with severity criteria (OR 3.52; 95%CI 1.07-11.90) were independently associated with the occurrence of C-PLA. In multivariate analysis, C-PLA was associated with a higher risk of recurrence (HR 3.08; 95%CI 1.38-11.22). Candida liver abscesses in non-neutropenic is a rare and severe disease. The high rate of recurrence should lead to discuss a more intensive treatment.


Morphology of the deciduous tusk (tush) of the African elephant (Loxodonta africana).
The tusk of the African elephant is preceded by a deciduous tooth generally known as the tush. Tushes from nine elephant fetuses and six calves younger than 1 year were exposed by dissection and described morphologically. All tushes consisted of a crown, root and pulpal cavity, the formation of which is completed soon after birth. They reached a maximum length of 5 cm, appeared not to erupt through the skin and were pushed aside and resorbed during enlargement of the distally located primordium of the tusk. Dental enamel, which covered the crown, could easily be removed and consisted of rods with an interwoven arrangement; the dentine-enamel junction was flat. Cellular cementum extended for variable distances over the crown and the dentine was tubular in nature. Although the tush apparently has no function, it provides the anlage and orientation for the development of its permanent successor.


Structure of a non-camelized human M12-V(H) domain at 1.5A resolution.
A non-camelized human V(H) domain has been crystallized through limited in vitro proteolysis of scFvM12 antibody fragment. The protease addition results in the complete degradation of the M12-V(L) domain, linker, and purification tags. The structure solved up to 1.5A resolution having good stereochemistry with a R(cryst) factor of 15.8% and R(free) factor of 19.7%. Dihedral angle values comparison of the first and the second complementarity-determining region (CDR) of M12-V(H) domain with an average values show a significant deviation; therefore, M12-V(H) domain structure indicates either the existence of a new canonical subclass or a link among the subclasses of canonical main-chain conformation in V(H)3 family. The presence of uncommon hydrogen bond between Ser-H50 and Tyr-H97 has pulling effect on CDR-H3 loop. The interface area buried by CDR-H3 loop indicates the partial coverage of the hydrophobic V(L)-V(H) interface. The isolated M12-V(H) domain was found soluble up to 0.35 mM. This result would be helpful in structure based designing of an isolated human single domain antibody fragments for biotechnological and pharmaceutical applications such as cancer.


Resistance patterns of Mycobacterium tuberculosis isolates from pulmonary tuberculosis patients in Nairobi.
In Kenya, which ranks thirteenth of 27 high tuberculosis burden countries, diagnosis is based on Ziehl-Neelsen staining alone and patients are treated without information on sensitivity patterns. This study aimed to determine resistance patterns of Mycobacterium tuberculosis isolated from pulmonary samples. Pulmonary tuberculosis patients in Nairobi were randomly sampled after informed consent and recruited into the study using a structured questionnaire. Specimens were cultured in liquid and solid media, and drug susceptibility tests were performed for first-line drugs including (isoniazid, rifampin, streptomycin, ethambutol and pyrazinamide). Eighty-six (30%) of 286 isolates were resistant to at least one of five antibiotics tested. Thirty-seven (30.2%) isolates were resistant to isoniazid; 15 (11.6%) to streptomycin; 13 (4.5%) to ethambutol; four (1.4%) to rifampin ; and 30 (10.4%) to pyrazinamide. Double resistance was seen as follows: four (1.4%) isolates were resistant to both isoniazid and pyrazinamide; four (1.4%) to streptomycin and isoniazid; and one (0.3%) to rifampin and streptomycin. Two isolates (0.7%) were multidrug resistant, and one was triple resistant with an additional resistance to ethambutol. Results also showed 88.7% of patients were below the age of 40 years, while 26.3% were HIV positive. The majority of the patients (66.5%) were unemployed or self-employed in small businesses, with 79.4% earning less than 100 USD per month. The high resistance observed in isoniazid, which is a first-line drug, could result in an increase in multidrug resistance unless control programs are strengthened. Poverty should be addressed to reduce infection rates.


History and instrumentation of laparoscopic surgery.
Equine surgery continues to benefit from advances in endoscopic surgical techniques. Much of what we have learned in equine endoscopy comes from techniques used in humans. Descriptions of the normal anatomy of the standing horse and advances in instrumentation have improved our ability to perform laparoscopic surgery in standing and dorsally recumbent horses. This article describes the history of laparoscopy and available instrumentation.


Brugada ECG pattern in hypertrophic cardiomyopathy: Brugada phenocopy or overlapping syndrome?
The term phenocopy indicates a condition that imitates one produced by a gene and is also used for acquired Brugada-like ECG manifestations. Cases of Brugada phenocopies are increasingly reported in literature and an international registry is ongoing. We describe two patients with Hypertrophic Cardiomyopathy (HCM) and Brugada ECG pattern. Both patients carried the same pathogenic splicing mutation in MYBPC3 gene (responsible for HCM) while no genetic mutation associated with Brugada Syndrome was identified. To the best of our knowledge, Brugada ECG pattern has been rarely reported in patients with HCM.


Results of subtotal parathyroidectomy in hemodialysis patients.
In 61 hemodialysis patients undergoing subtotal parathyroidectomy, there was a good correlation between the preoperative serum immunoreactive parathyroid hormone value (iPTH) and the weight of parathyroid tissue removed surgically (p less than or equal to 0.001). Postoperatively, iPTH decreased rapidly from an initial mean (+/- SD) of 2,928 +/- 1,600 muleq/ml and remained at 365 +/- 296 muleq/ml at last follow-up of patients still undergoing hemodialysis (normal, less than 50 muleq/ml). Of six patients who had recurrent hyperparathyroidism (10 percent of total), three required a second subtotal parathyroidectomy. Aluminum-related osteomalacia eventually developed in six patients with bone biopsy-proven hyperparathyroidism before parathyroidectomy. Nine patients with severe fracturing bone disease and hypercalcemia preoperatively but without clear evidence of hyperparathyroidism did not show a favorable response to subtotal parathyroidectomy (high mortality within 28 months, persistence of hypercalcemia, and symptomatic bone disease). Thus, subtotal parathyroidectomy can benefit patients with clearly established severe progressive hyperparathyroidism not responsive to medical therapy but is contraindicated in patients with low iPTH values and no bone biopsy evidence of severe hyperparathyroidism.


Cytological grading of breast carcinoma--a feasible proposition?
Fine needle aspiration cytology (FNAC) of the breast is widely used in the diagnosis of breast carcinoma. In some centres this is sometimes the only diagnostic procedure performed prior to definitive treatment. A grading system based on cytology would be helpful in the selection of patients for appropriate therapy. The aim of this study, therefore, was to devise such a system for grading breast carcinoma based on cytological features alone. The features assessed were the degree of cell clustering, nuclear pleomorphism, nuclear diameter, the presence of multiple, easily visible nucleoli and necrosis. Cytological features were compared to the histological grade of the tumours following excision. Discriminant analysis showed that the features with the closest correlation with histological grade were nuclear diameter, nuclear pleomorphism and the presence of nucleoli. A scoring system based on these three parameters enabled the classification of tumours into high and low cytological grades which showed a close correlation with histological grade.


Dose Modification in Biologic Therapy for Moderate to Severe Psoriasis: A Descriptive Analysis in a Clinical Practice Setting.
In biologic therapy, dose modification in carefully selected patients when psoriasis is in remission could reduce treatment costs and the risks associated with drug exposure. Observational, descriptive, crosssectional study, performed in January 2014, of 112 patients with moderate to severe psoriasis who had been on biologic therapy for at least 6 months. The therapeutic objective in all cases was to achieve and maintain a 75% reduction in Psoriasis Area and Severity Index (PASI 75). All the patients had started treatment with the standard regimen. During treatment, the dose had been reduced in patients who achieved the therapeutic objective and escalated in those who failed to respond adequately to standard doses. At the time of the study, 42.9% of the patients were receiving the standard dose, 50% were on a reduced dose, and 7.1% were on an escalated regimen. The agent with which the dose was most often reduced was adalimumab (57.7%), and the agents with which therapy was most often escalated were ustekinumab (17.9%) and infliximab (12.5%). Patients who received reduced doses had significantly longer-standing disease (P=.049) and longer treatment duration with the same biologic agent (P=.009). In the group that did not fulfill the criteria for dose reduction, the proportion of patients with psoriatic arthritis was significantly higher (P=.023). Cost savings were as follows: 21.5% with adalimumab, 13.8% with etanercept, .9% with ustekinumab, and .55% with infliximab. Patients with longer-standing disease and longer treatment duration with the same biologic agent were significantly more likely to be candidates for dose reduction. The proportion of patients with psoriatic arthritis was greater in the group of patients who did not fulfill the conditions for dose reduction. The overall cost saving achieved using the dose modification algorithm described in this study was 13%. Controlled studies are needed to define the profile of the patients best suited for dose reduction strategies without loss of treatment efficacy.


Maternal and perinatal outcomes of prolonged second stage of labour: a historical cohort study of over 51,000 women.
Prolonged second stage of labour has been associated with adverse maternal and perinatal outcomes. The maximum length of the second stage from full dilatation to birth of the baby remains controversial. Our aim was to determine whether extending second stage of labour was associated with adverse maternal and perinatal outcomes. A retrospective cohort study was conducted using routinely collected hospital data from 51592 births in Aberdeen Maternity Hospital between 2000 and 2016. The hospital followed the local guidance of allowing second stage of labour to extend by an hour compared to national guidelines since 2008 (nulliparous and parous). The increasing duration of second stage of labour was the exposure. Baseline characteristics, maternal and perinatal outcomes were compared between women who had a second stage labour of (a) ≤ 3 h and (b) > 3 h duration for nulliparous women; and (a) ≤ 2 h or (b) > 2 h for parous women. An additional model was run that treated the duration of second stage of labour as a continuous variable (measured in hours). All the adjusted models accounted for: age, BMI, smoking status, deprivation category, induced birth, epidural, oxytocin, gestational age, baby birthweight, mode of birth and parity (only for the final model). Each hourly increase in the second stage of labour was associated with an increased risk of obstetric anal sphincter injury (aOR 1.21 95% CI 1.16,1.25), having an episiotomy (aOR 1.48 95% CI 1.45, 1.52) and postpartum haemorrhage (aOR 1.27 95% CI 1.25, 1.30). The rates of caesarean and forceps delivery also increased when second stage duration increased (aOR 2.60 95% CI 2.50, 2.70, and aOR 2.44 95% CI 2.38, 2.51, respectively.) Overall adverse perinatal outcomes were not found to change significantly with duration of second stage on multivariate analysis. As the duration of second stage of labour increased each hour, the risk of obstetric anal sphincter injuries, episiotomies and PPH increases significantly. Women were over 2 times more likely to have a forceps or caesarean birth. The association between adverse perinatal outcomes and the duration of second stage of labour was less convincing in this study.


Lower extremity nerve blocks.
Lower extremity nerve blocks have not become as popular as upper extremity blocks for anesthesia; however, the use of lower extremity nerve blocks will become more widespread, as teaching programs are now providing more regional anesthesia experiences for their trainees so that the anesthesia provider will have the familiarity to use these blocks. To increase the enthusiasm among our surgical colleagues, we must begin to use these blocks for surgery, and if the block must be supplemented with local anesthetic or a light general anesthetic, we must educate them that the block is not a failure but a success, as it will provide analgesia after surgery in a method of multimodal pain control. Lower extremity nerve blocks will become more popular when it is realized that they are an effective way of increasing operating room efficiency. Because the block may be placed in an induction room, there is no induction or emergence in the operating room. Patients may be discharged without the need for pain medications, thus lowering the incidence of nausea postoperatively and decreasing PACU and discharge times.


n-3 fatty acids reduce proteinuria in patients with chronic glomerular disease.
Dietary supplementation with n-3 polyunsaturated fatty acids (n-3 PUFA) has been shown to reduce proteinuria in experimental models of renal diseases, but their potential role in the treatment of human renal disease is unknown. We administered n-3 PUFA in the form of triglycerides [with eicosapentaenoic (EPA)+docosahexaenoic (DHA) = 3 g/day into 4 patients] and of ethyl esters (EPA+DHA = 7.7 g/day) into 10 patients (one patient twice) with chronic glomerular disease (membranous glomerulonephritis and focal glomerular sclerosis), all diagnosed histologically. Serum albumin was > 2.4 g/dl and serum creatinine < 2.5 mg/dl in all patients. Treatment was given for periods of six weeks, followed by a prolonged follow-up for 27 weeks in 10 cases. Dietary supplementation with n-3 PUFA caused the expected reduction in platelet generation of thromboxane B2 (mean +/- SEM, from 490 +/- 70 ng/ml at baseline, to 342 +/- 147 ng/ml at 6 weeks, P < 0.05) of serum triglycerides (from 236 +/- 60 to 170 +/- 43, P < 0.01), and a prolongation of the bleeding time (from 5.8 +/- 0.4 min to 7.7 +/- 0.4 min, P < 0.01) in patients treated with ethyl esters. A modest but significant reduction in serum total cholesterol was noticed (from 275 +/- 27 to 252 +/- 24 mg/dl).(ABSTRACT TRUNCATED AT 250 WORDS)


Density dependent selection incorporating intraspecific competition 1. A haploid model.
A haploid model is introduced and analyzed in which intraspecific competition is incorporated within a density dependent framework. It is assumed that each genotype has a unique carrying capacity corresponding to the equilibrium population size when fixed for that type. Each genotypic fitness at a single multi-allelic locus is a function of a distinctive effective population size formed by adding the numbers of each genotype present, weighted by an intraspecific competition coefficient. As a result, the fitnesses depend upon the relative frequencies of the various genotypes as well as the total population size. Intergenotypic interactions can have a profound effect upon the outcome of the population. In particular, when the density effect of one individual upon another depends upon their respective genotypes, a unique stable interior equilibrium is possible in which all alleles are present. This stands in contrast to the purely density dependent haploid system in which the only possible stable state corresponds to fixation for the type with the highest carrying capacity. In the present model selective advantage is determined by a balance between carrying capacity and sensitivity to density pressures from other genotypes. Fixation for the genotype with the highest carrying capacity, for instance, will not be stable if it exerts a sufficiently weak competitive effect upon the other genotypes. In the diallelic case, maintenance of both alleles at a stable equilibrium requires that the net intragenotypic competition between individuals of like genotype be stronger than that between unlike types. As for purely density regulated systems, there may be no stable equilibria and/or regular and chaotic cycling may occur. The results may also be interpreted in terms of a discrete time model of interspecific competition with each haplotype representing a different species.


A Platelet/CMC coupled with offline UPLC-QTOF-MS/MS for screening antiplatelet activity components from aqueous extract of Danshen.
Platelets play crucial roles in thromboembolic and cardiovascular disease. The main platelets membrane receptors include adenosine diphosphate receptors, thrombin receptors, thromboxane prostanoid receptors and collagen receptors. In this study, a Platelet/CMC coupled with offline UPLC-QTOF-MS/MS system was built to screen antiplatelet activity components from aqueous extract of Danshen, which serve as an agent of antiplatelet aggregation in Traditional Chinese Medicine. Rosmarinic acid, lithospermic acid, salvianolic acid B, two isomers of salvianolic acid B, salvianolic acid C, salvianolic acid D and salvianolic acid H/I were identified as the potential antiplatelet activity components. Moreover, rosmarinic acid, lithospermic acid, salvianolic acid B, salvianolic acid C and danshensu were tested in platelet aggregation in vitro assay. The results suggested their retention time was closely related to the antiplatelet aggregation activities. This study provides a rapid, effective and novel method for screening the potential antiplatelet activity components from Chinese herb medicines.


Prediction of Microvascular Invasion in Combined Hepatocellular-Cholangiocarcinoma Based on Pre-operative Clinical Data and Contrast-Enhanced Ultrasound Characteristics.
The goal of the study described here was to define the predictive value of pre-operative clinical information and contrast-enhanced ultrasound (CEUS) imaging characteristics in combined hepatocellular-cholangiocarcinoma (CHC) patients with microvascular invasion (MVI). Seventy-six patients with pathologically confirmed CHC were enrolled in this study, comprising 18 patients with MVI-positive status and 58 with MVI-negative CHC nodules. The pre-operative clinical data and CEUS imaging features were retrospectively analyzed. Univariate and multivariate analyses were performed to identify the potential predictors of MVI in CHC. Recurrence-free survival (RFS) after hepatectomy was compared between patients with different MVI status using the log-rank test and Kaplan-Meier survival curves. Univariate analysis indicated that the following parameters of patients with CHC significantly differed between the MVI-positive and MVI-negative groups (p<0.05): tumor size, α-fetoprotein ≥400 ng/mL, enhancement patterns in arterial phase and marked washout during the portal venous phase on CEUS. On multivariate logistic regression analysis, only the CEUS characteristics of heterogeneous enhancement (odds ratio = 6.807; 95% confidence interval [CI]: 1.099, 42.147; p = 0.039) and marked washout (odds ratio = 4.380; 95% CI: 1.050,18.270; p = 0.043) were identified as independent predictors of MVI in CHC. The combination of the two risk factors in predicting MVI achieved a better diagnostic performance than each parameter alone, with an area under the receiver operating characteristic curve of 0.736 (0.622, 0.830). After hepatectomy, CHC patients with MVI exhibited earlier recurrence compared with those without MVI (hazard ratio = 1.859; 95% CI: 0.8699-3.9722, p = 0.046). The CEUS imaging features of heterogeneous enhancement in the arterial phase and marked washout during the portal venous phase were the potential predictors of MVI in CHC. Aside from that, CHC patients with MVI had an earlier recurrence rate than those without MVI after surgery.


PKC-MEK-MAPK-dependent signal transduction pathway mediates the stimulation of lysyl oxidase expression by serum and PDGF in rat aortic smooth muscle cells.
Lysyl oxidase (LO) plays a critical role in the stabilization and insolubilization of fibrous structural proteins of the extracellular matrix and has been implicated in the suppression of Ras-induced tumorigenesis. Several prior reports demonstrate that the expression of this catalyst is strongly influenced by a variety of effectors of cell function and is responsive to the growth state of fibrogenic cells. Using specific inhibitors of components of signal transduction pathways, the present study reveals that a PKC-MEK-MAPK-dependent pathway is critical to the enhanced expression of the LO gene in response to variations in the levels of the serum component of the growth medium and in response to platelet-derived growth factor (PDGF). PDGF is shown to be the major component of fetal bovine serum, which stimulates the activity of a LO promoter construct.


Heating treatment schemes for enhancing chelant-assisted phytoextraction of heavy metals from contaminated soils.
Recent research has shown that chelant-assisted phytoextraction approaches often require a high dosage of chelant applied to soil. The present study focused on optimization of phytoremediation processes to increase the phytoextraction efficiency of metals at reduced chelant applications. Pot experiments were carried out to investigate the effects of increased soil temperature on shoot uptake of heavy metals by corn (Zea mays L.) and mung bean (Vigna radiat L. Wilczek) from heavy metal-contaminated soils. After the application of S,S-ethylenediaminedisuccinic acid or ethylenediaminetetra-acetic acid, soils were exposed to high temperatures (50 or 80 degrees C) for 3 h, which significantly increased the concentration of heavy metals in shoots. The heating treatment 2 d after the chelant addition resulted in higher concentrations of metals compared with those treatments 2 d before or simultaneously with the chelant application. Irrigation with 100 degrees C water 2 d after the chelant addition, or irrigation with 100 degrees C chelant solutions directly, also resulted in significantly higher phytoextraction of metals in the two crops compared with 25 degrees C chelant solutions. In addition, a novel application method to increase soil temperature using underground polyvinyl chloride tubes would increase the chelant-assisted extraction efficiency of Cu approximately 10- to 14-fold in corn and fivefold in mung bean compared with those nonheating treatments. In a field experiment, increasing soil temperature 2 d after chelant addition also increased the shoot Cu uptake approximately fivefold compared with those nonheating treatments. This new technique may represent a potential, engineering-oriented approach for phytoremediation of metal-polluted soils.


Choosing the appropriate section thickness in the melamine embedding technique.
When biological materials are infiltrated by a water-soluble melamine resin and hardened, they become as hard as glass. This is a prerequisite for extreme thin-sectioning. In this paper, the structural information from unsupported transparent thin sections of beef liver catalase, calf thymus DNA, horse spleen ferritin, insect muscle and rat microtubules is compared to that of normal thin sections. While ferritin molecules (12 nm diameter), microtubule subunits (8 nm long axis) and catalase crystals (8 nm subunit diameter) appear to become mechanically damaged in a 10 nm section (as measured by resectioning), DNA-molecules (3 nm diameter) are satisfactorily preserved during sectioning. Remarkably, for electron phase contrast imaging of unstained cross-sectioned insect muscle, a minimum section thickness of about 30-40 nm is required.


A new cause for difficulty in seeing at night.
A syndrome is described in which patients complain of inability to see at night, although their rod and cone thresholds are normal. In such persons the rods, when dark-adapted, elevate cone flicker thresholds by 2 log units or more, and this condition is the basis of their complaint. Patients are often thought to be hysterics or to have congenital stationary night blindness.


Sigma-binding site ligands inhibit K+ currents in rat locus coeruleus neurons in vitro.
Biological actions of novel sigma1- and sigma2-selective binding site ligands (trishomocubanes: 4-azahexacyclo [220.127.116.11.(2,6).0(3,10).0(5,).0(8,11)]dodecanes), and the reference ligands, 1,3-di(2-tolyl)-guanidine (DTG), haloperidol, (+)-pentazocine and dextromethorphan, were studied in rat locus coeruleus neurons using intracellular and whole-cell patch clamp recordings. High concentrations of trishomocubanes produced small inward currents and affected some parameters of action potential waveforms suggesting modest potency to inhibit ionic conductances underlying action potentials. Sigma-ligands produced large inward currents in the presence of mu-opioid, alpha2-adrenoceptor and ORL1 receptor agonists. These reversed polarity near the K+ equilibrium potential, suggesting that sigma-ligands act as ligand activated K+-channel blockers or interfere with the coupling between these receptors and K+-channels. However, no correlation was found between binding affinities at sigma1- or sigma2-binding sites and potency to inhibit K+-currents, suggesting that these effects on K+-channels are not directly related to occupancy of sigma binding sites.


Dose-Dependent Suppression of Gonadotropins and Ovarian Hormones by Elagolix in Healthy Premenopausal Women.
Elagolix is a nonpeptide, oral gonadotropin-releasing hormone (GnRH) antagonist being developed for sex-hormone-dependent diseases in women. We evaluated the pharmacokinetics and pharmacodynamics of elagolix. This study was a randomized, double-blind, placebo-controlled, multiple-ascending dose study in 45 healthy premenopausal women at a research unit. Elagolix [150 mg once daily or 100, 200, 300, or 400 mg twice daily (BID)] or placebo was administered for 21 days. Main outcome measures were elagolix pharmacokinetics, suppression of gonadotropics [follicle-stimulating hormone (FSH), luteinizing hormone (LH)] and ovarian hormones [estradiol (E2), progesterone (P)], and adverse events. Elagolix was rapidly absorbed after oral dosing, reaching maximum concentrations at 1.0 to 1.5 hours, with a half-life of 4 to 6 hours. FSH, LH, and E2 were suppressed within hours of elagolix administration on day 1. Dose-dependent suppression of E2 was observed, with maximum suppression achieved with elagolix 200 mg BID. Dose-dependent suppression of FSH and LH was also observed, with maximal or near-maximal suppression achieved at 300 mg BID and 200 mg BID, respectively. At elagolix doses ≥100 mg BID, P concentrations remained at anovulatory levels throughout 21 days of dosing. The most frequently reported adverse events were headache and hot flush. Elagolix administration allows for modulation of gonadotropin and ovarian hormone concentrations, from partial suppression at lower doses to nearly full suppression at higher doses. The results of this study provide a rationale for elagolix dose selection for treatment of sex hormone-dependent diseases in women.


Transcription-terminating mutation in telethonin causing autosomal recessive muscular dystrophy type 2G in a European patient.
A 27-year-old woman of Moldavian origin presented at the age of 15 with progressive proximal limb weakness and painful cramps in her calf muscles. Clinical examination revealed prominent muscle weakness in proximal muscles of the lower extremities and distal anterior compartment of legs, and mild weakness in shoulder girdle muscles. In addition, she had marked calf hypertrophy, muscle atrophy involving the anterior and posterior compartments of the thighs, and the distal anterior compartment of legs, as well as mild scapular winging and hyperlordosis. A muscle biopsy taken from the biceps brachii showed mild dystrophic changes, absent vacuoles, and abundant lobulated fibers. Immunofluorescence and Western blot assays demonstrated complete telethonin deficiency. Molecular analysis revealed a homozygous Trp25X mutation in the telethonin (TCAP) gene resulting in termination of transcription at an early point. Four families from Brazil with telethonin deficiency have previously been reported and classified as LGMD2G, but the actual frequency of this disease is unknown. With this current identification of a case outside the Brazilian population, telethonin mutation-associated LGMD should be considered worldwide.


Separation and identification of botanical insecticide 7-hydroxycoumarin and its biological activity against Aphis craccivora and Culex pipiens pallens.
The root powder of Stellera chamaejasme was extracted with chloroform. The crude extract obtained was chromatographed with silica gel columns many times. One pure natural product was obtained. It was identified as 7-hydroxycoumarin. Laboratory bioassay showed that it had a good insecticidal activity against Aphis craccivora and Culex pipiens pallens. Its median lethal concentrations (LC50) (24 h after treatment) were 19.56 and 0.435 mg L(-1), respectively. The results of regressive and correlative analysis demonstrated that the correlations were very significant between concentration and efficacy. The corresponding correlative coefficients were 0.9686 and 0.9712, respectively. This is the first report of the insecticidal activity of 7-hydroxycoumarin. It may represent a new class of insecticides.


PackHelix: a tool for helix-sheet packing during protein structure prediction.
The three-dimensional structure of a protein is organized around the packing of its secondary structure elements. Predicting the topology and constructing the geometry of structural motifs involving α-helices and/or β-strands are therefore key steps for accurate prediction of protein structure. While many efforts have focused on how to pack helices and on how to sample exhaustively the topologies and geometries of multiple strands forming a β-sheet in a protein, there has been little progress on generating native-like packings of helices on sheets. We describe a method that can generate the packing of multiple helices on a given β-sheet for αβα sandwich type protein folds. This method mines the results of a statistical analysis of the conformations of αβ(2) motifs in protein structures to provide input values for the geometric attributes of the packing of a helix on a sheet. It then proceeds with a geometric builder that generates multiple arrangements of the helices on the sheet of interest by sampling through these values and performing consistency checks that guarantee proper loop geometry between the helices and the strands, minimal number of collisions between the helices, and proper formation of a hydrophobic core. The method is implemented as a module of ProteinShop. Our results show that it produces structures that are within 4-6 Å RMSD of the native one, regardless of the number of helices that need to be packed, though this number may increase if the protein has several helices between two consecutive strands in the sequence that pack on the sheet formed by these two strands.


Hypomorphic phenotype of Foxn1 gene-modified rats by CRISPR/Cas9 system.
The Foxn1 gene is known as a critical factor for the differentiation of thymic and skin epithelial cells. This study was designed to examine the phenotype of Foxn1-modified rats generated by the CRISPR/Cas9 system. Guide-RNA designed for first exon of the Foxn1 and mRNA of Cas9 were co-injected into the pronucleus of Crlj:WI zygotes. Transfer of 158 injected zygotes resulted in the birth of 50 offspring (32 %), and PCR identified five (10 %) as Foxn1-edited. Genomic sequencing revealed the deletion of 44 or 60 bp from and/or insertion of 4 bp into the Foxn1 gene in a single allele. The number of T-cells in the peripheral blood lymphocytes of mutant rats decreased markedly. While homozygous deleted mutant rats had no thymus, the mutant rats were not completely hairless and showed normal performance in delivery and nursing. Splicing variants of the indel-mutation in the Foxn1 gene may cause hypomorphic allele, resulting in the phenotype of thymus deficiency and incomplete hairless. In conclusion, the mutant rats in Foxn1 gene edited by the CRISPR/Cas9 system showed the phenotype of thymus deficiency and incomplete hairless which was characterized by splicing variants.


Hemodynamics and intracardiac conduction after operative repair of tetralogy of Fallot.
Electrophysiologic studies were performed in 47 children aged 3 to 18 years, 15 of whom had cardiac arrhythmias 1 to 15 years after repair of tetralogy of Fallot. Six exhibited sinus or atrioventricular nodal dysfunction, 8 had ventricular extrasystoles, and 1 had supraventricular tachycardia. Hemodynamic and electrophysiologic data were obtained at postoperative catheterization. Although electrophysiologic responses were abnormal in a proportion of both the children with and those without arrhythmia, hemodynamic values were similar. Three of 6 children with impaired sinus impulse generation or atrioventricular nodal conduction had a prolonged A-H interval, and in 3 Wenckebach heart block developed at low pacing rates. Ventricular ectopic rhythm was not associated with any particular abnormality of basic intracardiac conduction intervals. Thus, arrhythmias and conduction abnormalities are not consistently related to residual right ventricular hypertension. Abnormalities in electrophysiologic function are common after repair of tetralogy of Fallot in patients with sinus rhythm and may have prognostic implications for these patients.


Ultrastructural and physicochemical characterization of the hepatitis C virus recovered from the serum of an agammaglobulinemic patient.
Hepatitis C virus (HCV) morphology and physicochemical properties remain unclear because HCV usually circulates in a complexed form in association with immunoglobulins. In the present work, we were interested in the characterization of HCV particles derived from the serum of an anti-HCV negative/HCV RNA positive agammaglobulinemic patient suffering from chronic type C hepatitis. Physicochemical properties of the virus particles were determined by serum centrifugation on a 10-60% isopycnic sucrose density gradient. HCV RNA quantified by bDNA was found in a major peak at density 1.13 g/ml and in a minor peak at densities 1.05-1.07 g/ml. By electron microscopy, 45 nm large core-like particles were found at the 1.13 g/ml density while 60 nm large virus-like particles similar to other members of the Flaviviridae family were visualized at the 1.06-1.07 g/ml densities. This confirms some studies reporting the low density of HCV as compared to other members of the Flaviviridae family.


Loss of Endometrial Sodium Glucose Cotransporter SGLT1 is Detrimental to Embryo Survival and Fetal Growth in Pregnancy.
Embryo implantation requires a hospitable uterine environment. A key metabolic change that occurs during the peri-implantation period, and throughout early pregnancy, is the rise in endometrial glycogen content. Glycogen accumulation requires prior cellular uptake of glucose. Here we show that both human and murine endometrial epithelial cells express the high affinity Na+-coupled glucose carrier SGLT1. Ussing chamber experiments revealed electrogenic glucose transport across the endometrium in wild type (Slc5a1 +/+) but not in SGLT1 deficient (Slc5a1 -/-) mice. Endometrial glycogen content, litter size and weight of offspring at birth were significantly lower in Slc5a1 -/- mice. In humans, SLC5A1 expression was upregulated upon decidualization of primary endometrial stromal cells. Endometrial SLC5A1 expression during the implantation window was attenuated in patients with recurrent pregnancy loss when compared with control subjects. Our findings reveal a novel mechanism establishing adequate endometrial glycogen stores for pregnancy. Disruption of this histiotrophic pathway leads to adverse pregnancy outcome.


My Personal Journey from the Fascination for Phages to a Tumor-Inducing Fungal Pathogen of Corn.
My path in science began with a fascination for microbiology and phages and later involved a switch of subjects to the fungus Ustilago maydis and how it causes disease in maize. I will not provide a review of my work but rather focus on decisive findings, serendipitous, lucky moments when major advances made the U. maydis-maize system what it is now-a well-established model for biotrophic fungi. I also want to share with you the joy of finding the needle in a haystack at the very end of my scientific career, a fungal structure likely used for effector delivery, and how we were able to translate this into a potential application in agriculture.


Effect of oral administration of Pinellia ternata, Zingiberis rhizoma and their mixture on the efferent activity of the gastric branch of the vagus nerve in the rat.
The effect of taste stimulation of Pinellia temata, Zingiberis rhizoma and their mixture on the efferent activity of the gastric branch of the vagus nerve was observed in the anesthetized rat. Taste stimulation by Pinellia ternata (50 mg/ml, 10 min) resulted in a suppression in vagal gastric nerve activity. On the contrary, stimulation by Zingiberis rhizoma (50 mg/ml, 10 min) caused a facilitation in efferent activity. The mixture of Pinellia ternata and Zingiberis rhizoma (5:1, 50 mg/ml, 10 min stimulation) demonstrated no suppressive effect on gastric nerve activity. These observations indicate that it is reasonable to prescribe Pinellia ternata with Zingiberis rhizoma in traditional Japanese medicine to prevent suppressive effect of the taste of Pinellia ternata on gastric function.


High expression of IGFBP2 is associated with chemoresistance in adult acute myeloid leukemia.
Insulin-like growth factor (IGF) signaling plays an important role in many tumors and overexpression of IGF Binding Protein (IGFBP) 2 has been associated with adverse outcome in childhood leukemia. Here, we evaluated IGFBP2 mRNA expression and its prognostic implications in 99 adult acute myeloid leukemia (AML) patients by quantitative real-time RT-PCR. High IGFBP2 was associated with a high incidence of primary resistant disease (IGFBP2 high 65%, IGFBP2 low 32%; P=0.02) and was independently predictive for therapy resistance [OR 3.6 (95% CI 1.2-11); P=0.02] in multivariate analyses. Gene-expression profiling revealed an up-regulation of genes implicated in leukemogenesis (MYB, MEIS1, HOXB3, HOXA9) and genes associated with adverse outcome (ERG, WT1) in patients with high IGFBP2 expression. Thus, our data suggest a role of IGFBP2 and IGF signaling in chemoresistance of AML. Patients with high IGFBP2 expression might benefit from molecular therapies targeting the IGF pathway.


The essential mitochondrial protein Erv1 cooperates with Mia40 in biogenesis of intermembrane space proteins.
The proteins of the mitochondrial intermembrane space (IMS) are encoded by nuclear genes and synthesized on cytosolic ribosomes. While some IMS proteins are imported by the classical presequence pathway that involves the membrane potential deltapsi across the inner mitochondrial membrane and proteolytic processing to release the mature protein to the IMS, the import of numerous small IMS proteins is independent of a deltapsi and does not include proteolytic processing. The biogenesis of small IMS proteins requires an essential mitochondrial IMS import and assembly protein, termed Mia40. Here, we show that Erv1, a further essential IMS protein that has been reported to function as a sulfhydryl oxidase and participate in biogenesis of Fe/S proteins, is also required for the biogenesis of small IMS proteins. We generated a temperature-sensitive yeast mutant of Erv1 and observed a strong reduction of the levels of small IMS proteins upon shift of the cells to non-permissive temperature. Isolated erv1-2 mitochondria were selectively impaired in import of small IMS proteins while protein import pathways to other mitochondrial subcompartments were not affected. Small IMS precursor proteins remained associated with Mia40 in erv1-2 mitochondria and were not assembled into mature oligomeric complexes. Moreover, Erv1 associated with Mia40 in a reductant-sensitive manner. We conclude that two essential proteins, Mia40 and Erv1, cooperate in the assembly pathway of small proteins of the mitochondrial IMS.


Inhibitory effect of free fatty acids on plasma protein binding of disopyramide in haemodialysis patients.
The plasma (or serum) protein binding of disopyramide (DSP) in five haemodialysis patients was studied using an ultrafiltration technique. There was an increase in the free fraction of DSP in the plasma on dialysis days in comparison to the levels on interdialysis days, which was associated with an elevation of the free fatty acid levels in the plasma together with the increase of the free fraction of DSP. The inhibitory effect of free fatty acids on DSP binding in an in vitro study was enhanced in proportion to their concentrations, and was shown to be due to competition at one binding site by experiments with oleic acid as a representative displacer. Certain endogenous organic acids, such as indoxyl sulphate, 2-hydroxyhippuric acid and hippuric acid, which are characteristically elevated in chronic renal failure, scarcely affected the protein binding of DSP. The findings indicate that free DSP should be monitored in patients with elevated plasma free fatty acid levels, such as those on haemodialysis therapy.


Crucial Role of the Chaperonin GroES/EL for Heterologous Production of the Soluble Methane Monooxygenase from Methylomonas methanica MC09.
Methane is a widespread energy source and can serve as an attractive C1 building block for a future bioeconomy. The soluble methane monooxygenase (sMMO) is able to break the strong C-H bond of methane and convert it to methanol. The high structural complexity, multiplex cofactors, and unfamiliar folding or maturation procedures of sMMO have hampered the heterologous production and thus biotechnological applications. Here, we demonstrate the heterologous production of active sMMO from the marine Methylomonas methanica MC09 in Escherichia coli by co-synthesizing the GroES/EL chaperonin. Iron determination, electron paramagnetic resonance spectroscopy, and native gel immunoblots revealed the incorporation of the non-heme diiron centre and homodimer formation of active sMMO. The production of recombinant sMMO will enable the expansion of the possibilities of detailed studies, allowing for a variety of novel biotechnological applications.