Pioglitazone attenuates TGF-beta(1)-induction of fibronectin synthesis and its splicing variant in human mesangial cells via activation of peroxisome proliferator-activated receptor (PPAR)gamma.
The peroxisome proliferator-activated receptor (PPAR)gamma is expressed not only in adipose tissue but also in macrophages/monocytes and plays important roles in acute/chronic inflammation. Transforming growth factor (TGF)-beta is a common pathogenic indicator of sclerosis because it induces the accumulation of extracellular matrix (ECM) in the glomerular mesangium of the kidney. Among components of the ECM, fibronectin (FN) is an acute reactant in inflammation, and isoforms of it produced by splicing of gene variants appear during abnormal conditions such as wound healing. In this study, we examined the effects of pioglitazone, a PPARgamma agonist, on TGF-beta(1)-induced FN synthesis in cultured mesangial cells using RT-PCR and Western blot analysis. We also analyzed its splicing variant, extra domain (ED) A, containing FN (EDA(+)FN). TGF-beta(1) enhanced the production of both FN and EDA(+) FN and down-regulated PPARgamma expression. Pioglitazone reversed both these effects of TGF-beta(1). These findings suggest that PPARgamma activation by pioglitazone may affect the TGF-beta(1)-induced FN accumulation observed in the glomerular mesangium in cases of glomerulosclerosis, although further in vivo experiments are needed to evaluate this inference.


Membrane potential changes during IgE-mediated histamine release from rat basophilic leukemia cells.
The membrane potential of rat basophilic leukemia cells (RBL-2H3 cell line) has been determined by monitoring the distribution of the lipophilic [3H] tetraphenylphosphonium cation between the cells and the extracellular medium. By this method, the determined potential of these cells, passively sensitized with IgE, is -93 +/- 5 mV (mean +/- SEM, interior negative). Almost 40% of this membrane potential is rapidly collapsed upon the addition of the proton carrier, carbonyl cyanide p-trifluoromethoxyphenyl hydrazone (FCCP). It is suggested that the FCCP-sensitive fraction of the total membrane potential results from the accumulation of this cation by the mitochondria, which maintains a negative membrane potential. Thus, the resting plasma membrane potential of these cells equals -55 +/- 6 mV. During the process of immunological stimulation by antibodies directed against cell membrane bound IgE, the membrane potential decreases. Moreover, there is a correlation between the extent of degranulation of the cells and the depolarization. It is concluded that in common with other secretory systems, depolarization of the plasma membrane is involved in the stimulus-secretion coupling of the histamine secreting RBL cells.


Neural correlates of hierarchical predictive processes in autistic adults.
Bayesian theories of autism spectrum disorders (ASD) suggest that atypical predictive mechanisms could underlie the autistic symptomatology, but little is known about their neural correlates. Twenty-six neurotypical (NT) and 26 autistic adults participated in an fMRI study where they performed an associative learning task in a volatile environment. By inverting a model of perceptual inference, we characterized the neural correlates of hierarchically structured predictions and prediction errors in ASD. Behaviorally, the predictive abilities of autistic adults were intact. Neurally, predictions were encoded hierarchically in both NT and ASD participants and biased their percepts. High-level predictions were following activity levels in a set of regions more closely in ASD than NT. Prediction errors yielded activation in shared regions in NT and ASD, but group differences were found in the anterior cingulate cortex and putamen. This study sheds light on the neural specificities of ASD that might underlie atypical predictive processing.


Enamel matrix derivative as adjunctive to non-surgical periodontal therapy: a systematic review and meta-analysis of randomized controlled trials.
To assess the potential additional benefit of the local application of enamel matrix derivative (EMD) on the clinical outcomes following non-surgical periodontal therapy (NSPT) (steps 1 and 2 periodontal therapy). A systematic literature search was performed in several electronic databases, including Medline/PubMed, Embase, The Cochrane Register of Central Trials (CENTRAL), LILACS, and grey literature. Only randomized controlled clinical trials (RCTs) were eligible for inclusion. Clinical attachment level (CAL) change (primary outcome), probing pocket depth (PPD), and bleeding on probing (BoP) reductions (secondary outcomes) were evaluated. The Cochrane Risk of Bias tool (RoB 2.0) was used to assess the quality of the included trials. Weighted mean differences (WMDs) and 95% confidence intervals (CIs) between test and control sites were estimated using a random-effect model for amount of mean CAL and PPD change. Six RCTs were included for the qualitative analysis, while data from 4 studies were used for meta-analysis. Overall analysis of CAL gain (3 studies) and PPD reduction (4 studies) presented WMD of 0.14 mm (p = 0.74; CI 95% - 0.66; 0.94) and 0.46 mm (p = 0.25; CI 95% - 0.33; 1.26) in favor of NSPT + EMD compared to NSPT alone respectively. Statistical heterogeneity was found to be high in both cases (I2 = 79% and 87%, respectively). Within their limitations, the present data indicate that the local application of EMD does not lead to additional clinical benefits after 3 to 12 months when used as an adjunctive to NSPT. However, due to the high heterogeneity among the studies, additional well-designed RCTs are needed to provide further evidence on this clinical indication for the use of EMD. The adjunctive use of EMD to NSPT does not seem to additionally improve the clinical outcomes obtained with NSPT alone.


Downregulation of miR-26b attenuates early brain injury induced by subarachnoid hemorrhage via mediating the KLF4/STAT3/HMGB1 axis.
Early brain injury (EBI) refers to early-onset secondary complications that occur after subarachnoid hemorrhage (SAH), and associated with high rate of disability and mortality. Recent investigations have indicated microRNA-26b (miR-26b) as a biomarker in the progression of SAH. Accordingly, the present study was designed to elucidate the role of miR-26b in influencing EBI following SAH and the downstream mechanisms. Firstly, SAH rat models and neuron injury models were developed to assess the effect of miR-26b on EBI-like symptoms and subsequent inflammation. Dual-luciferase reporter gene assay was further implemented to evaluate the binding of miR-26b to its putative target gene STAT3. Loss-of-function and rescue experiments were performed to assess the functionality of miR-26b-mediated STAT3 in both models. miR-26b was found to target KLF4 and negative-modulate its expression, whereby aggravating EBI and inflammatory response in SAH rat models and stimulating hemoglobin-induced apoptosis in astrocytes. On the other hand, silencing of miR-26b reversed these changes in SAH rat models and hemoglobin (Hb)-induced astrocytes. miR-26b could further activate STAT3 via down-regulation of KLF4. Furthermore, KLF4 knockdown up-regulated HMGB1 to aggravate EBI following SAH. Collectively, our findings highlighted the ameliorative effect of miR-26b inhibition on EBI in SAH and the possible mechanism associated with the KLF4/STAT3/HMGB1 axis.


Characterization of binding kinetics of [3H]Tyr-D-Arg2-Phe-Sar4 at opioid receptors.
The dermorphin-derived, MOP receptor-selective tetrapeptide Tyr-D-Arg2-Phe-Sar4 (TAPS) exhibits a high antinociceptive potency and stimulates respiration in rats. The receptor binding kinetics of [3H]TAPS were investigated using crude calf thalamic membrane preparations. Saturation studies showed binding of [3H]TAPS at two binding sites (0.4 and 3.2 nM). In the presence of MgSO4, [3H]TAPS binding occurred with high affinity at a single site only. The high-affinity binding component was reduced following the addition of K2-EDTA. The increase of the apparent dissociation constant was due to an enhanced dissociation rate (P<0.05), while association rates remained unchanged. Addition of 5'-guanylylimidodiphosphate (Gpp(NH)p) resulted in a reduction of affinity which was augmented in the presence of Na+. Thus, [3H]TAPS, depending on the presence of divalent cations, serves as ligand at two MOP receptor binding sites in the calf thalamus, which may represent distinct affinity states of the same receptor or receptor subtypes thereof.


Real-time continuous estimation of gas exchange by dual oximetry.
We designed a ventilation-perfusion index (VQI) to estimate venous admixture (Qsp/Qt) in a real-time fashion by simultaneous pulse and pulmonary artery oximetry in 17 patients with acute respiratory failure. Changes in Qsp/Qt were produced by altering the level of continuous positive airway pressure. VQI correlated well with Qsp/Qt (r = 0.78). This contrasts with the poor correlation found between Qsp/Qt and the commonly used oxygen tension based indices such as PaO2/FIO2 (r = -0.51), PaO2/PAO2 (r = 0.47), and PAO2-PaO2 (r = 0.23). The use of dual oximetry to derive a VQI appears to be a reliable and accurate method for real-time assessment of pulmonary gas exchange in patients with acute respiratory failure.


Comparative toxicokinetics of 2,3-14C-and 1-14C-acrylonitrile in the rat.
The tissue distribution, elimination and covalent binding of 2,3-14C-and 1-14C-acrylonitrile (VCN) were studies in male Sprague-Dawley rats given an oral dose of 46.5 mg kg-1. Exhalation of unchanged VCN, 14CO2 and H14CN was monitored at selected intervals. Only 5% of the total dose administered was recovered was unchanged VCN. Rate given 2,3-14C-VCN exhaled only 2% of 14C activity was 14CO2 and none was recovered as H14CN, whereas rats given 1-14C-VCN exhaled about 12% of 14C activity as 14CO2 and 0.5% as H14CN. In the initial 24 h, 40% of radioactivity from 1-14C-VCN appeared in urine, while 60% was recovered in the urine of rats given 2,3-14C-VCN. The red blood cells retained significant amounts of radioactivity from both the compounds for more than 10 days after administration, whereas the 14C activity in plasma declined sharply. The highest level of radioactivity from both compounds was recovered in the gastrointestinal tract. In liver, kidney, brain, spleen, adrenal, lung and heart tissues the unbound percent radioactivity decreased, while irreversible percent covalent binding to macromolecules in relation to total increased concomitantly. Subcellular fractionation of the tissues showed that most of the covalently bound radioactivity was distributed in non-cytosolic fractions. As compared to 1-14C-VCN administered animals, the percentage of covalent binding of 2,3-14C-VCN was significantly higher even 72 h after dosing. The relationship between covalent binding and acrylonitrile toxicity is discussed.


Utility of enzymes from Fibrobacter succinogenes and Prevotella ruminicola as detergent additives.
In this study, we investigated the application of cellulase and protease purified from rumen bacteria as detergent additives. Cellulase and protease were purified from the rumen cellulytic bacteria Fibrobacter succinogenes S85, and Prevotella ruminicola 23, respectively. An inhibitor test indicated that the purified protease belongs to the category of serine proteases and metalloproteases. Both the enzymes were effective at a high temperature (50 degrees C) and neutral pH (pH 7-8), but the protease activity increased with the increase in temperature and pH. The purified protease was treated with ten types of surfactants/detergents; it was found to retain over 60% of its activity in the presence of anionic and nonionic detergents. The cellulose plus protease combination was still effective after treatment with Triton X-100 and Tween 80, but the residual activity was low after treatment with Tween 20 than that after treatment with other nonionic detergents. Washing tests indicated that enzyme addition produced no significant improvement in the removal of grass stains, but individual enzyme addition in surfactants/detergents, especially in nonionic detergents, could improve the washing performance of the detergents by improving its ability to remove blood stains. This suggested that the surfactant/detergent class, enzyme properties, and the mixing ratio of ingredients should be considered simultaneously to enhance the washing performance.


Maternal secretin ameliorates obesity by promoting white adipose tissue browning in offspring.
Our knowledge of the coordination of intergenerational inheritance and offspring metabolic reprogramming by gastrointestinal endocrine factors is largely unknown. Here, we showed that secretin (SCT), a brain-gut peptide, is downregulated by overnutrition in pregnant mice and women. More importantly, genetic loss of SCT in the maternal gut results in undesirable phenotypes developed in offspring including enhanced high-fat diet (HFD)-induced obesity and attenuated browning of inguinal white adipose tissue (iWAT). Mechanistically, loss of maternal SCT represses iWAT browning in offspring by a global change in genome methylation pattern through upregulation of DNMT1. SCT functions to facilitate ubiquitination and degradation of DNMT1 by activating AMPKα, which contributes to the observed alteration of DNMT1 in progeny. Lastly, we showed that SCT treatment during pregnancy can reduce the development of obesity and improve glucose tolerance and insulin resistance in offspring of HFD-fed females, suggesting that SCT may serve as a novel biomarker or a strategy for preventing metabolic diseases.


Molecular mechanisms of nonablative fractionated laser resurfacing.
Nonablative fractionated laser resurfacing improves the texture of treated skin, but little is known about the molecular mechanisms that underlie clinical improvements. We sought to examine and quantify the time course and magnitude of dermal matrix changes that occur in response to nonablative fractionated laser resurfacing, with the dual goals of better understanding the molecular mechanisms that underlie clinical improvements and of gaining knowledge that will enable evidence-based treatment parameter optimization. Twenty patients (mean age 58 years) with photodamaged skin were focally treated on dorsal forearms with a nonablative fractionated laser. Serial skin samples were obtained at baseline and at various times after treatment. Biopsies were examined with real-time polymerase chain reaction technology and immunohistochemical techniques. Laser treatment resulted in an initial inflammatory response as indicated by statistically significant induction of proinflammatory cytokines (interleukin-1β and tumour necrosis factor-α). This was followed by substantial increases in levels of several matrix metalloproteinases and later by significant induction of type I collagen. Dermal remodelling was noted with both low and high microbeam energy treatment parameters. Nonablative fractionated laser resurfacing induces a well-organized wound-healing response that leads to substantial dermal remodelling and collagen induction. Surprisingly, only minimal differences were observed between lower and higher microbeam energy settings. These data suggest that lower microbeam energy/higher microbeam density treatment parameters, which are generally better tolerated by patients, may yield dermal changes similar to those that result from higher microbeam energy/lower microbeam density treatment parameters.


Is there equity of patient health outcomes across models of general practice in Aotearoa New Zealand? A national cross-sectional study.
Primary care in Aotearoa New Zealand is largely delivered by general practices, heavily subsidised by government. Te Tiriti o Waitangi (1840) guarantees equal health outcomes for Māori and non-Māori, but differences are stark and longstanding. Seven models of primary care have evolved. We hypothesised that patient health outcomes would differ between models of care; and that Māori, Pacific peoples and those living in material deprivation would have poorer outcomes from primary care. We conducted a cross-sectional study of patient-level data from national datasets and practices, at 30 September 2018, using multilevel mixed effects regression analyses (patients clustered within practices). Primary outcomes, considered to be measures of unmet need for primary care, were polypharmacy (≥ 65 years), HbA1c testing in adults with diabetes, childhood immunisations (6 months), ambulatory sensitive hospitalisations (0-14, 45-64 years) and emergency department attendances. Explanatory variables adjusted for patient and practice characteristics. Equity, by model of care, ethnicity and deprivation, was assumed if they showed no significant association with patient outcomes. Patient characteristics included: age, ethnicity, deprivation, multi-morbidity, first specialist assessments and practice continuity. Practice characteristics included: size, funding and doctor continuity. Clinical input (consultations and time with nurses and doctors) was considered a measure of practice response. The study included 924 general practices with 4,491,964 enrolled patients. Traditional practices enrolled 73% of the population, but, on average, the proportion of Māori, Pacific and people living with material deprivation was low in any one Traditional practice. Patients with high health needs disproportionately enrolled in Māori, Pacific and Trust/NGO practices. There were multiple associations between models of care and patient health outcomes in fully adjusted regressions. No one model of care out-performed others across all outcomes. Patients with higher health need received more clinical input but this was insufficient to achieve equity in all outcomes. Being a Māori or Pacific patient, or living in material deprivation, across models of care, remained associated with poorer outcomes. Model-level associations with poor patient outcomes suggest inequity in measures that might be used to target investment in primary care.


Genome-scale CRISPR screening at high sensitivity with an empirically designed sgRNA library.
In recent years, large-scale genetic screens using the CRISPR/Cas9 system have emerged as scalable approaches able to interrogate gene function with unprecedented efficiency and specificity in various biological contexts. By this means, functional dependencies on both the protein-coding and noncoding genome of numerous cell types in different organisms have been interrogated. However, screening designs vary greatly and criteria for optimal experimental implementation and library composition are still emerging. Given their broad utility in functionally annotating genomes, the application and interpretation of genome-scale CRISPR screens would greatly benefit from consistent and optimal design criteria. We report advantages of conducting viability screens in selected Cas9 single-cell clones in contrast to Cas9 bulk populations. We further systematically analyzed published CRISPR screens in human cells to identify single-guide (sg) RNAs with consistent high on-target and low off-target activity. Selected guides were collected in a novel genome-scale sgRNA library, which efficiently identifies core and context-dependent essential genes. We show how empirically designed libraries in combination with an optimized experimental design increase the dynamic range in gene essentiality screens at reduced library coverage.


Key molecules involved in the Th17/Treg balance are associated with the pathogenesis of reflux esophagitis and Barrett's esophagus.
Reflux esophagitis (RE) impairs the squamous epithelium that normally lines the esophagus, and contributes to the replacement of the damaged squamous lining by the intestinal metaplasia of Barrett's esophagus (BE), which is considered as a precursor of esophageal adenocarcinoma. This study aimed to investigate the changes in the balance of Th17/Treg and the related key molecules in the pathogenesis of RE and BE and evaluate the diagnostic and predictive value of the molecules in patients with these diseases. The proportions of Th17 and Treg in RE and BE patients were estimated using flow cytometric analysis. Key molecules involving in the Th17/Treg balance, including RORγt, Foxp3, IL-17, IL-6, IL-10, and TGF-β, were measured using quantitative real-time PCR (qRT-PCR) and ELISA analyses. The diagnostic and predictive value of the Th17/Treg ratio and its key regulators was evaluated using a receiver operating characteristic assay (ROC). In addition, the Spearman correlation analysis explored the relationship between the Th17/Treg ratio and the clinical characteristics. An increased ratio of Th17/Treg was observed in RE and BE compared with the normal controls, and the proportion of Th17/Treg in BE was further increased compared with RE patients. Moreover, the expression levels of RORγt, IL-17, IL-6, and TGF-β were elevated, while the levels of Foxp3 and IL-10 were reduced in patients when compared to the controls. Compared with the RE groups, the levels of IL-17 were significantly higher in BE patients, while the Foxp3 levels were significant decreased. In addition, the Th17/Treg ratio also showed high diagnostic significance and considerable predictive value for the clinical outcomes in patients with RE and BE. The balance of Th17/Treg was impaired in patients with RE and BE. Th17/Treg may be involved in the development of both RE and BE through regulating the release of inflammatory cytokines, but the concrete mechanisms maybe different in the two diseases. The imbalance of Th17/Treg ratio and the related key molecules had a certain clinical diagnosis and prediction potential for RE and BE.


Distribution of cGMP-dependent and cGMP-independent Ca(2+)-activated Cl(-) conductances in smooth muscle cells from different vascular beds and colon.
In the present patch-clamp study we have, for the first time, shown the tissue distribution of a recently characterized cGMP-dependent Ca(2+)-activated Cl(-) conductance [18] in smooth muscle cells freshly isolated from different regions: aorta, pulmonary artery, tail artery, femoral artery, femoral vein, middle cerebral artery, renal artery, portal vein, superior mesenteric artery, mesenteric small artery and colon. The cGMP-dependent Cl(-) conductance has properties distinct from those of the 'classical' Ca(2+)-activated Cl(-) conductances; their different sensitivities to niflumic acid and zinc were here utilized to distinguish them. They were found to be co-expressed in different patterns in smooth muscle cells of different origins. The cGMP-dependent conductance was greater in myocytes from cerebral artery and femoral vein and was greater in the renal artery, aorta, mesenteric small artery, femoral artery and the superior mesenteric artery. The presence of the cGMP-dependent Ca(2+)-activated Cl(-) current in smooth muscle cells isolated from the colon demonstrates that this conductance is not limited to the vasculature. The 'classical' Ca(2+)-activated Cl(-) conductance was strongly expressed in smooth muscle cells from the portal vein and the tail artery, and noticeably higher in the pulmonary artery.


Lichenoid dermatitis: a clinicopathologic and immunopathologic review of sixty-two cases.
We reviewed 62 cases (26 men and 36 women, median age 55 years) of previously diagnosed lichenoid dermatitis and correlated the histologic and immunodermatologic findings with the clinical diagnosis and course. The clinical diagnostic groups were contact dermatitis (6 cases), drug eruption (14), lupus erythematosus (6), lichen planus (3), cutaneous T cell lymphoma (3), chronic dermatitis or neurodermatitis (19), and miscellaneous dermatologic disorders (11). With the use of individual histologic features, a correlation with specific clinical conditions was possible, and the clinical groups of dermatitis, drug eruption, lupus erythematosus or lichen planus, and T cell infiltration could be selected histologically. Direct immunofluorescence studies were of most help in the diagnosis of lupus erythematosus and atypical lichen planus, but these studies should be performed in all cases involving lichenoid inflammation. Although previously the nonspecific term lichenoid dermatitis was used to classify the histologic features in these cases, we found that by careful correlation of histologic, immunodermatologic, and clinical features, we were often able to provide a specific diagnosis.


Mental illness, family and networks in a London borough: two cases studied by an anthropologist.
The situation of crisis undergone by a family when one of its members is admitted to psychiatric hospital is a privileged moment for anthropological investigation of the relationship between the individual and society. Social rules are being experienced, tested and explained through a crisis that is apparently localized in the individual although the whole family is involved in various ways. In this paper a detailed ethnography of such crises in two families in a London Borough will illuminate two levels of analysis: (a) the cultural frameworks which give meaning to these events (b) the network mobilized by the families in an attempt to cope with the situation. It is my hypothesis that the following analysis of the patterns of coping exhibited by these two families will provide us with two models of the way in which crises of mental illness are dealt with. In the first family, a 'sociocentric' model of explanation is utilised and is associated with a segregated conjugal role relationship and an amplified movement of the nuclear family in relation to its network. In the second case, an 'egocentric' model is utilised in order to understand the crisis. This is associated with a joint role--relationship between husband and wife and a contracted movement in relation to the network. The hypothesis, exemplified in specific situations of social drama, stresses the movement of a social unit (the family), and indicates its dynamic character, the existence of different choices and distinct conceptions of the idea of the relationship between the person and his immediate social environment--the family. This paper is particularly significant since there exist very few detailed studies of urban contexts in Western societies containing data about the meaning of mental illness in the social environment in which it occurs.


The prevalence, characteristics, and related factors of pressure injury in medical staff wearing personal protective equipment against COVID-19 in Turkey: A multicenter cross-sectional study.
The aim of this study was to determine the prevalence of device-related pressure injury (DRPI) related to the use of personal protective equipment (PPE) among healthcare professionals during the COVID-19 pandemic and the associated risk factors effective in the development of DRPI. This descriptive, cross-sectional and correlational study was conducted with 1465 healthcare professionals working in healthcare institutions in Eastern Turkey during the COVID-19 pandemic. The study data were collected by means of an online anonymous survey questioning the prevalence, characteristics, and associated factors of DRPI in the first week of April 2021, using the snowball sampling method. Number, percentage, arithmetic mean, Chi-Square, and regression analysis were used for the evaluation of the study data. The prevalence of DRPI due to the use of PPE use among healthcare professionals was calculated to be 60.5%. Of the developed DRPIs, 79.5% were stage 1, and the most frequent anatomical locations of DRPI were the bridge of the nose (30.2%), behind the ears (24.6%), and cheeks (20.8%). The logistic regression analysis revealed that male gender, age <35 years, being a physician and nurse, prolonged duration of PPE use (>4 h), working in a high-risk clinic (COVID-19 clinic and COVID-19 intensive care unit), and sweating during the use of PPE were predictive factors for the development of DRPI (p < 0.05). The prevalence of DRPI due to PPE use among healthcare workers was quite high. Gender, age, occupation, long duration of PPE use, working in a high-risk clinic, and sweating during the use of PPE were found to be risk factors in the development of DRPI.


First complete genome sequence of garlic virus X infecting Allium sativum- G282 from India.
The present report communicates the first full genome sequencing of the Garlic virus X from northern India. The total genome size of Garlic virus X (MK503771) reported in this study is 8458 bp ssRNA. The full genome sequence analysis showed the close relationship of Garlic virus X from India to that of from China, Korea, Australia and Spain. The full genome sequence based study of Indian Garlic virus X reveals the geographical relationship of this virus in India and global origin which may assists in development of control strategy for this virus.


Evaluation of the humpback scaphoid by computed tomography in the longitudinal axial plane of the scaphoid.
The deformity known as the humpback scaphoid is difficult to visualize on radiographs. Three patients were studied by imaging with computerized axial tomography (CT scan) in the true longitudinal axis of the scaphoid. This projection demonstrates the anatomy, allowing assessment of fractures, the humpback deformity, and the shape of the scaphoid after grafting.


STING/NF-κB/IL-6-Mediated Inflammation in Microglia Contributes to Spared Nerve Injury (SNI)-Induced Pain Initiation.
Innate immune response acts as the first line of host defense against damage and is initiated following the recognition of pathogen-associated molecular patterns (PAMPs). For double-stranded DNA (dsDNA) sensing, interferon gene stimulator (STING) was discovered to be an integral sensor and could mediate the immune and inflammatory response. Selective STING antagonist C-176 was administered and pain behaviors were assessed following spared nerve injury (SNI)-induced neuropathic pain. The level of serum dsDNA following neuropathic pain was assessed using Elisa analysis. STING signaling pathway, microglia activation, and proinflammatory cytokines were assessed by qPCR, western blots, Elisa, and immunofluorescence staining. STING agonist DMXAA was introduced into BV-2 cells to assess the inflammatory response in microglial cells. dsDNA was significantly increased following SNI and STING/TANK-binding kinase 1 (TBK1)/nuclear factor-kappa B (NF-κB) pathway was activated in vivo and vitro. Early but not the late intrathecal injection of C-176 attenuated SNI-induced pain hypersensitivity, microglia activation, proinflammatory factors, and phosphorylated JAK2/STAT3 in the spinal cord dorsal horn, and the analgesic effect of C-176 was greatly abolished by recombinant IL-6 following SNI. We provided evidence clarifying dsDNA mediated activation of microglia STING signaling pathway, after which promoting expression of proinflammatory cytokines that are required for hyperalgesia initiation in the spinal cord dorsal horn of SNI model. Further analysis showed that microglial STING/TBK1/NF-κB may contribute to pain initiation via IL-6 signaling. Pharmacological blockade of STING may be a promising target in the treatment of initiation of neuropathic pain.


Differences in the effectiveness of EDTA to induce SCEs and chromosomal aberrations in CHO and Allium cepa chromosomes.
The chelating agent EDTA was able to produce chromosome aberrations (CA) in CHO cells when it was administered simultaneously with BrdUrd (2 x 10(-5) M), without any concomitant effect on the yield of sister chromatid exchanges (SCEs). Root meristematic cells of Allium cepa did not show any type of CA when they were treated with different doses of EDTA (with or without BrdUrd 10(-4) M) while the SCE frequency was increased in a dose-dependent fashion. These effects of EDTA have not been previously reported. It is suggested that deprivation of divalent cations (Ca(+)+/Mg(+)+) probably play an important role in DNA replication and repair processes.


An observational study of clinical recurrence in patients with interstitial lung disease related to the antisynthetase syndrome.
To describe the clinical characteristics and risk factors of clinical recurrence in interstitial lung disease related to antisynthetase syndrome (ARS-ILD). Patients diagnosed as ARS-ILD in Nanjing Drum Tower Hospital between January 2015 and November 2020 were retrospectively analyzed. Clinical information and treatment course were reviewed. The primary endpoint was the disease recurrence, and the secondary point was mortality. Univariate and multivariable Cox regression analyses were performed to identify risk factors for recurrence. Totally, 132 patients with ARS-ILD received immunomodulation treatment from diagnosis. During follow-ups, sixty-nine patients showed recurrence, with a recurrency rate yielding 52.3%. The median duration from treatment initiation to recurrence was 11 (5-18) months. The median tapering course in the recurrence group was 8 (3-12.5) months, which was significantly shorter than the 16 (10-32) months in the no-recurrence group (p < 0.001). Fifty-eight patients experienced recurrence when the glucocorticoids (GC) dose dropped to 10 (9.375-15) mg/day. Twelve patients discontinued GC with a median treatment course of 11.5 (8-16.75) months, and 11 patients developed recurrence after discontinuing GC for 3 (1-4) months. Twelve patients died, with a mortality rate of 9.1%, and recurrence was not associated with increased mortality. The adjusted multivariate analysis showed that age, increased serum lactate dehydrogenase (LDH) level, relatively shorter tapering duration, and inappropriate GC discontinuation were associated with recurrence. Recurrence of ARS-ILD was common during medication intensity reduction. Age, LDH, medication tapering duration, and discontinuation were risk factors for recurrence. Further efforts to reduce recurrence should take into consideration of these factors. Key Points • Recurrence is observed commonly with a recurrency rate 52.3% in patients with interstitial lung disease related to antisynthetase syndrome (ARS-ILD) when glucocorticoids (GC) tapering or discontinuation. • Age, increased serum lactate dehydrogenase (LDH) level, medication tapering duration, and GC discontinuation were identified to be significantly associated with the recurrence of ARS-ILD.


Resolving an unnoticed diversity within the Schistura robertsi species complex (Teleostei: Nemacheilidae) using molecules and morphology.
The Schistura robertsi species complex is a group of freshwater fish inhabiting streams in southeast Myanmar as well as in western and southern Thailand. In southern Thailand, the distribution exceeds the biogeographically important 'Surat Thani - Krabi line'. The complex is believed to include five described and one undescribed species, but monophyly and systematics of the group have never been studied explicitly. The present study aims to resolve the number of species within the Schistura robertsi group as well as their distribution areas and phylogenetic relations. We analysed mitochondrial and nuclear sequence data of 86 specimens from 47 localities and 18 morphological characters of 193 specimens. The phylogenetic analyses revealed the S. robertsi complex to be monophyletic and to be composed of ten major lineages. Six of them correspond to the known described or undescribed species, but another four newly identified clades reveal the existence of an overlooked diversity within the group. All genetic lineages are statistically highly supported and all are morphologically diagnosable, suggesting that they represent distinct species. The distribution areas of several clades overlap, the cases of direct co-occurrence show no sign of hybridisation.


Terabyte-scale supervised 3D training and benchmarking dataset of the mouse kidney.
The performance of machine learning algorithms, when used for segmenting 3D biomedical images, does not reach the level expected based on results achieved with 2D photos. This may be explained by the comparative lack of high-volume, high-quality training datasets, which require state-of-the-art imaging facilities, domain experts for annotation and large computational and personal resources. The HR-Kidney dataset presented in this work bridges this gap by providing 1.7 TB of artefact-corrected synchrotron radiation-based X-ray phase-contrast microtomography images of whole mouse kidneys and validated segmentations of 33 729 glomeruli, which corresponds to a one to two orders of magnitude increase over currently available biomedical datasets. The image sets also contain the underlying raw data, threshold- and morphology-based semi-automatic segmentations of renal vasculature and uriniferous tubules, as well as true 3D manual annotations. We therewith provide a broad basis for the scientific community to build upon and expand in the fields of image processing, data augmentation and machine learning, in particular unsupervised and semi-supervised learning investigations, as well as transfer learning and generative adversarial networks.


Prognostic Assessment of Fractional Flow Reserve in Different Strata in Patients with Coronary Artery Disease.
There are limited real-world data on the clinical course of untreated coronary lesions according to their functional severity. To evaluate the 5-year clinical outcomes of patients with revascularized lesions with fractional flow reserve (FFR) ≤ 0.8 and patients with non-revascularized lesions with FFR > 0.8. The FFR assessment was performed in 218 patients followed for up to 5 years. Participants were classified based on FFR into ischemia group (≤ 0.8, intervention group, n = 55), low-normal FFR group (> 0.8-0.9, n = 91), and high-normal FFR group (> 0.9, n = 72). The primary endpoint was major adverse cardiac events (MACEs), a composite of death, myocardial infarction, and need for repeat revascularization. The significance level was set at 0.05; therefore, results with a p-value < 0.05 were considered statistically significant. Most patients were male (62.8%) with a mean age of 64.1 years. Diabetes was present in 27%. On coronary angiography, the severity of stenosis was 62% in the ischemia group, 56.4% in the low-normal FFR group, and 54.3% in the high-normal FFR group (p<0.05). Mean follow-up was 3.5 years. The incidence of MACEs was 25.5%, 13.2%, and 11.1%, respectively (p=0.037). MACE incidence did not differ significantly between the low-normal and high-normal FFR groups. Patients with FFR indicative of ischemia had poorer outcomes than those in non-ischemia groups. There was no difference in the incidence of events between the low-normal and high-normal FFR groups. Long-term studies with a large sample size are needed to better assess cardiovascular outcomes in patients with moderate coronary stenosis with FFR values between 0.8 and 1.0.


The use of a wearable cardioverter defibrillator during postmastectomy radiotherapy as an alternative for relocation of the pulse generator.
We describe a case in which a breast cancer patient with an implantable cardioverter-defibrillator (ICD) was treated with postmastectomy followed by axillary radiation therapy. The ICD was partly within the target volume and according to the treatment planning system would receive a maximum dose of about 36 Gy. Postoperatively, the patient had a wound infection needing surgical intervention and 3 months of antibiotics. Therefore, it was decided not to relocate the ICD but instead to switch off the shock function of the ICD for the entire radiation therapy course. The ICD was successfully substituted with a wearable cardioverter defibrillator (WCD), which the patient wore during the 3-week radiation therapy period until 2 weeks after the end of treatment. The ICD function was monitored from the start of the treatment until 6 months after treatment. In situations in which an ICD needs to be switched off semipermanently, a WCD can be used as alternative. The patient in this report tolerated the WCD well. Despite the high dose received by the ICD, the device and its battery continued to function normally.


Control and coding of pupil size by hypothalamic orexin neurons.
Brain orexin (hypocretin) neurons are implicated in sleep-wake switching and reward-seeking but their roles in rapid arousal dynamics and reward perception are unclear. Here, cell-specific stimulation, deletion and in vivo recordings revealed strong correlative and causal links between pupil dilation-a quantitative arousal marker-and orexin cell activity. Coding of arousal and reward was distributed across orexin cells, indicating that they specialize in rapid, multiplexed communication of momentary arousal and reward states.


Chemometrics-assisted spectroscopic methods for rapid analysis of combined anti-malarial tablets.
Combination of piperaquine (PQ) (320mg) and dihydroartemisinin (DHA) (40 mg) is an anti-malarial formulation, which is recommended by World Health Organization (WHO). Simultaneous analysis of PQ and DHA can be problematic due to the lack of chromophores or fluorophores in DHA molecule. Whereas PQ possesses strong UV absorption and it presents in 8 times of DHA contents in the formulation. In this study, two spectroscopic methods, Fourier transform infrared (FTIR) and Raman spectroscopy, were developed for the determination of both drugs in combined tablets. The FTIR and Raman spectra were recorded in the attenuate total reflectance (ATR) and scattering modes, respectively. The original and pretreated spectra from FTIR and handheld-Raman were subjected to Unscrambler® program to construct partial least squares regression (PLSR) model comparing with references values obtained from high performance liquid chromatography (HPLC)-UV method. The optimal PLSR models of PQ and DHA from FTIR spectroscopy were obtained from orthogonal signal correction (OSC) pretreatment at the wavenumbers 400-1,800 cm-1 and 1,400-4,000 cm-1, respectively. For Raman spectroscopy of PQ and DHA, the optimal PLSR models were obtained from standard normal variate (SNV) pretreatment at the wavenumbers 1,200-2,300 cm-1 and OSC pretreatment at the wavenumber 400-2,300 cm-1, respectively. Determination of PQ and DHA in tablets from the optimum model was compared with HPLC-UV method. Results were not significantly different at 95% confidence limit (p-value >0.05). The chemometrics-assisted spectroscopic methods were fast (1-3 min), economical and less labor intensive. Moreover, the handheld Raman spectrometer is portable and can be utilized for onsite analysis to facilitate the detection of counterfeit or substandard drugs at ports of entry.


Tracing the Evolution of Human Gene Regulation and Its Association with Shifts in Environment.
As humans populated the world, they adapted to many varying environmental factors, including climate, diet, and pathogens. Because many of these adaptations were mediated by multiple noncoding variants with small effects on gene regulation, it has been difficult to link genomic signals of selection to specific genes, and to describe the regulatory response to selection. To overcome this challenge, we adapted PrediXcan, a machine learning method for imputing gene regulation from genotype data, to analyze low-coverage ancient human DNA (aDNA). First, we used simulated genomes to benchmark strategies for adapting PrediXcan to increase robustness to incomplete data. Applying the resulting models to 490 ancient Eurasians, we found that genes with the strongest divergent regulation among ancient populations with hunter-gatherer, pastoralist, and agricultural lifestyles are enriched for metabolic and immune functions. Next, we explored the contribution of divergent gene regulation to two traits with strong evidence of recent adaptation: dietary metabolism and skin pigmentation. We found enrichment for divergent regulation among genes proposed to be involved in diet-related local adaptation, and the predicted effects on regulation often suggest explanations for known signals of selection, for example, at FADS1, GPX1, and LEPR. In contrast, skin pigmentation genes show little regulatory change over a 38,000-year time series of 2,999 ancient Europeans, suggesting that adaptation mainly involved large-effect coding variants. This work demonstrates that combining aDNA with present-day genomes is informative about the biological differences among ancient populations, the role of gene regulation in adaptation, and the relationship between genetic diversity and complex traits.


Platelet adhesion and aggregate formation controlled by immobilised and soluble VWF.
It has been demonstrated that von Willebrand factor (VWF) mediated platelet-endothelium and platelet-platelet interactions are shear dependent. The VWF's mobility under dynamic conditions (e.g. flow) is pivotal to platelet adhesion and VWF-mediated aggregate formation in the cascade of VWF-platelet interactions in haemostasis. Combining microfluidic tools with fluorescence and reflection interference contrast microscopy (RICM), here we show, that specific deletions in the A-domains of the biopolymer VWF affect both, adhesion and aggregation properties independently. Intuitively, the deletion of the A1-domain led to a significant decrease in both adhesion and aggregate formation of platelets. Nevertheless, the deletion of the A2-domain revealed a completely different picture, with a significant increase in formation of rolling aggregates (gain of function). We predict that the A2-domain effectively 'masks' the potential between the platelet glycoprotein (GP) Ib and the VWF A1-domain. Furthermore, the deletion of the A3-domain led to no significant variation in either of the two functional characteristics. These data demonstrate that the macroscopic functional properties i.e. adhesion and aggregate formation cannot simply be assigned to the properties of one particular domain, but have to be explained by cooperative phenomena. The absence or presence of molecular entities likewise affects the properties (thermodynamic phenomenology) of its neighbours, therefore altering the macromolecular function.


Innate apoptotic immunity: the calming touch of death.
Apoptotic cell death is an essential and highly ordered process that contributes to both the development and the homeostasis of multicellular organisms. It is associated with dramatic biochemical and cell biological events within the dying cell, including fragmentation of the nucleus and the redistribution of intracellular proteins and membrane lipids. It has long been apparent that phagocytic clearance of the cell corpse is an integral part of the apoptotic process; apoptotic clearance also may be essential in tissue homeostasis. During the cell death process, apoptotic cells acquire new cell surface determinants for specific recognition by responder phagocytes and suppression of immune responsiveness. Recent studies indicate that these determinants are well conserved throughout metazoan evolution; remarkably, their recognition shows no species-specific restriction. Professional and non-professional phagocytes recognize and respond to apoptotic cells similarly, notably with the immediate-early transcriptional repression of a variety of specific genes including those encoding inflammatory cytokines. Secondary responses following engulfment of the apoptotic corpse, utilizing several distinct mechanisms, enhance and sustain this apoptotic suppression. In this review, we highlight the central role of apoptotic cells in innate homeostatic regulation of immunity.


Metabonomic modeling of drug toxicity.
Global metabolic profiling (metabonomics/metabolomics) has shown particular promise in the area of toxicology and drug development. In both preclinical screening and mechanistic exploration, metabolic profiling can offer rapid, noninvasive toxicological information that is robust and reproducible, with little or no added technical resources to existing studies in drug metabolism and toxicity. In this review, the study design and analytical technology required for metabonomics are discussed, along with key examples of how fundamental questions in drug development can be addressed. Strategies for metabonomic data analysis in toxicity assessment are detailed in both principle and practice, together with a description of toxicologically relevant metabolic biomarkers. Extended into the assessment of efficacy and toxicity in the clinic, metabonomics may prove crucial in making personalized therapy and pharmacogenomics a reality.


Encryption chain based on measurement result and its applications on semi-quantum key distribution protocol.
This study proposes a new encoding method, also known as an encryption chain based on the measurement result. Then, using the encryption chain to propose a unitary-operation-based semi-quantum key distribution protocol (SQKD) protocol. In the existing SQKD protocols, semi-quantum environments adopt a round-trip transmission strategy. In round-trip transmission, the classical participant must resend the received photons to the quantum participant after implementing local operations. Therefore, round-trip transmissions are vulnerable to Trojan horse attacks. Hence, the classical participant must be equipped with a photon number splitter and an optical wavelength filter device against Trojan horse attacks. This is illogical for semi-quantum environments because the burden on the classical participant is significantly increased as it involves the prevention of Trojan horse attacks. The proposed SQKD protocol is congenitally immune to Trojan horse attacks and involves no extra hardware because it is designed based on a one-way transmission as opposed to a round-trip transmission. When compared to the existing SQKD protocols, the proposed SQKD protocol provides the best qubit efficiency, and classical participants only require two quantum capabilities, which enhance its practicability. Moreover, the proposed SQKD protocol is free from collective attacks, Trojan horse attacks, and intercept-resend attacks. Thus, the proposed scheme is more efficient and practical than the existing SQKD protocols.


Ruptured pseudoaneurysm of the aorta with encapsulated mediastinal abscess after coronary artery bypass grafting.
Rupture of pseudoaneurysm of the aorta with chronic active mediastinitis after cardiac surgery is a catastrophic complication. We report a case of one-stage operation of urgent repair for ruptured pseudoaneurysm under deep hypothermic circulatory arrest, debridement of mediastinum, and omental transfer, with an uneventful postoperative course.


Diagnosis of anal sphincter defects by three-dimensional transperineal ultrasound in women with anal incontinence.
To assess the utility of transperineal three-dimensional (3D) ultrasound for diagnosing anal sphincter defects and evaluating the function of the anal canal in women with anal incontinence. The study subjects were 13 women with anal incontinence. Symptoms of fecal incontinence were assessed by Wexner score. The anal canal of each woman was examined ultrasonically with both a convex transperineal 3D scanner and a radial transanal scanner to compare the accuracy of the two approaches for diagnosis of anal sphincter defects. The anorectal angle and the length of the anal canal were also measured by utilizing the functionality of the transperineal 3D ultrasound. The mean age was 58.9 ± 14.9 years (±SD), and the mean Wexner score was 8.4 ± 5.6. In terms of ultrasound diagnosis of anal sphincter defects, the two methods showed consistent results in each woman. The length of the portion where both the internal and external anal sphincters were intact was significantly correlated with the Wexner score, whereas the total length of the anal canal was not. Less invasive transperineal 3D ultrasound provides accurate evaluation of the internal and external anal sphincters in women with anal incontinence, and the method is potentially useful for detection of anal sphincter abnormalities.


Severe Versus Very Severe Pediatric Obstructive Sleep Apnea Outcomes After Adenotonsillectomy.
Adenotonsillectomy (AT) is generally considered the first line treatment for pediatric patients with obstructive sleep apnea (OSA). Pediatric patients with severe OSA have worse outcomes after AT than patients with milder OSA. It is currently unclear if this group of higher morbidity patients should be subdivided further. This study investigates patients with severe pediatric OSA to determine if there are differences in postsurgical outcomes based on initial severity of sleep disordered breathing, medical comorbidities, or demographic factors. Retrospective cohort study at a single tertiary referral center. Patients aged 2-18 who underwent polysomnogram (PSG) from October 2012 to January 2019, had an apnea-hypopnea index (AHI) >10, and subsequently underwent AT were identified using a filter through electronic medical record. A total of 112 patients underwent both pre- and postoperative PSG. Bivariate analysis was conducted via Pearson chi-square test. Univariate and multivariate analyses via binary logistic and multinomial linear regressions were performed using SPSS. Of the 112 patients included in this study, 68 patients were identified as having severe OSA (AHI = 10-20) and 44 as having very severe OSA (AHI > 20). Very severe OSA patients were significantly less likely to be cured of sleep disordered breathing or have their OSA reduced to mild OSA. Obese patients were found to have less reduction in AHI after AT. The postsurgical outcomes of patients with severe and very severe OSA are significantly different indicating that patients traditionally categorized as having severe OSA may need to be further subcategorized. 4 Laryngoscope, 132:1855-1860, 2022.


Breathlessness rating type influences respiratory behavior during hypercapnia in the rebreathing test.
The online rating of perceived breathlessness in a rebreathing test (RT) is a new and powerful technique to analyze the psychological and physiological process related to the mechanisms of breathlessness. The aim of the present study was to assess the influence of rating type on respiratory sensation and behavior during repeated hypercapnic exposures. Sixty-one healthy women performed three rebreathing trials. Half of them were auditorily cued to rate breathlessness intensity every 12 s (CR, n=30), the other half was instructed to rate breathlessness each time a change in breathlessness intensity was perceived (UCR, n=31). The ventilatory response was assessed simultaneously. The CR group yielded an earlier threshold in tidal volume (V(T)) than the UCR group. Only in the UCR group did changes occur across rebreathing trials: the V(T) threshold decreased, the respiratory rate (RR) threshold increased, and the sensitivity of the RR increased. The type of breathlessness rating influences respiratory behavior in the RT. The short-interval cued rating seems to be the best option for a standardized application of this test in research on panic disorder.


Calcium, mitochondria and apoptosis studied by fluorescence measurements.
Among the many unsolved problems of calcium signalling, the role of calcium elevations in apoptotic and necrotic cell death has been a focus of research in recent years. Evidence has been presented that calcium oscillations can effectively trigger apoptosis under certain conditions and that dysregulation of calcium signalling is a common cause of cell death. These effects are regularly mediated through calcium signal propagation to the mitochondria and the ensuing mitochondrial membrane permeabilization and release of pro-apoptotic factors from mitochondria to the cytoplasm. The progress in this area depended on the development of (1) fluorescent/luminescent probes, including fluorescent proteins that can be genetically targeted to different intracellular locations and (2) the digital imaging technology, fluorescence-activated cell sorting and fluorescent high throughput approaches, which allowed dynamic measurements of both [Ca2+] in the intracellular compartments of interest and the downstream processes. Fluorescence single cell imaging has been the only possible approach to resolve the cell-to-cell heterogeneity and the complex subcellular spatiotemporal organization of the cytoplasmic and mitochondrial calcium signals and downstream events. We outline here fluorometric and fluorescence imaging protocols that we set up for the study of calcium in the context of apoptosis.


Sequential detection of Salmonella typhimurium and Bacillus anthracis spores using magnetoelastic biosensors.
Multiple phage-based magnetoelastic (ME) biosensors were simultaneously monitored for the detection of different biological pathogens that were sequentially introduced to the measurement system. The biosensors were formed by immobilizing phage and 1mg/ml BSA (blocking agent) onto the magnetoelastic resonator's surface. The detection system included a reference sensor as a control, an E2 phage-coated sensor specific to S. typhimurium, and a JRB7 phage-coated sensor specific to B. anthracis spores. The sensors were free standing during the test, being held in place by a magnetic field. Upon sequential exposure to single pathogenic solutions, only the biosensor coated with the corresponding specific phage responded. As the cells/spores were captured by the specific phage-coated sensor, the mass of the sensor increased, resulting in a decrease in the sensor's resonance frequency. Additionally, non-specific binding was effectively eliminated by BSA blocking and was verified by the reference sensor, which showed no frequency shift. Scanning electron microscopy was used to visually verify the interaction of each biosensor with its target analyte. The results demonstrate that multiple magnetoelastic sensors may be simultaneously monitored to detect specifically targeted pathogenic species with good selectivity. This research is the first stage of an ongoing effort to simultaneously detect the presence of multiple pathogens in a complex analyte.


Melatonin Improves Skeletal Muscle Structure and Oxidative Phenotype by Regulating Mitochondrial Dynamics and Autophagy in Zücker Diabetic Fatty Rat.
Obesity-induced skeletal muscle (SKM) inflexibility is closely linked to mitochondrial dysfunction. The present study aimed to evaluate the effects of melatonin on the red vastus lateralis (RVL) muscle in obese rat models at the molecular and morphological levels. Five-week-old male Zücker diabetic fatty (ZDF) rats and their age-matched lean littermates (ZL) were orally treated either with melatonin (10 mg/kg body weight (BW)/24 h) (M-ZDF and M-ZL) or non-treated (control) (C-ZDF and C-ZL) for 12 weeks. Western blot analysis showed that mitochondrial fission, fusion, and autophagy were altered in the C-ZDF group, accompanied by reduced SIRT1 levels. Furthermore, C-ZDF rats exhibited depleted ATP production and nitro-oxidative stress, as indicated by increased nitrites levels and reduced SOD activity. Western blotting of MyH isoforms demonstrated a significant decrease in both slow and fast oxidative fiber-specific markers expression in the C-ZDF group, concomitant with an increase in the fast glycolytic fiber markers. At the tissue level, marked fiber atrophy, less oxidative fibers, and excessive lipid deposition were noted in the C-ZDF group. Interestingly, melatonin treatment partially restored mitochondrial fission/fusion imbalance in the RVL muscle by enhancing the expression of fission (Fis1 and DRP1) markers and decreasing that of fusion (OPA1 and Mfn2) markers. It was also found to restore autophagy, as indicated by increased p62 protein level and LC3BII/I ratio. In addition, melatonin treatment increased SIRT1 protein level, mitochondrial ATP production, and SOD activity and decreased nitrites production. These effects were associated with enhanced oxidative phenotype, as evidenced by amplified oxidative fiber-specific markers expression, histochemical reaction for NADH enzyme, and muscular lipid content. In this study, we showed that melatonin might have potential therapeutic implications for obesity-induced SKM metabolic inflexibility among patients with obesity and T2DM.


Open stone surgery in the treatment of bilateral complex renal stones with left infected hydronephrosis: A case report.
With the strong development of percutaneous nephrolithotomy as well as other less invasive procedures, the indication of open surgery for the treatment of nephrolithiasis has been significantly reduced and is only applied in selective cases. A 55-year-old male was admitted to the Department of Urology due to fever and left flank pain. Clinical examination and imaged studies reveal staghorn calculi in the right kidney and infected hydronephrosis with multiple stones on the left side. The stone removal surgery was performed in 2 sessions, left first - right after. The two operations came out with no peri- or post-operative complication, no blood transfusion. The two reasons for the indication of open surgery instead of percutaneous nephrolithotomy were i/ the stone's sizes were very large and very hard, and ii/ the multi-tract nephrolithotomy increased the risks of blood transfusion and parenchymal's damage. Also, a long-time and difficult nephroscopy was not the good choice for the left-infected hydronephrosis. Open stone surgery is still a good alternative approach for kidney complex stone burden, especially infected hydronephrosis.


Getting key players to work together and defending against diversion--Arizona.
In this report, the author traces the development of the offensive and defensive strategies employed by various antitobacco coalitions and organizations in Arizona. Further, he describes key principles and identifies directions for states soon to embark on tobacco control initiatives.


A high molecular weight inhibitor of Ca2+ -dependent neutral protease in rat brain.
An endogenous, heat-stable inhibitor of high mol. wt (approximately 3x10(5)) was found to be present in rat brain, which inhibited Ca2+-dependent neutral protease specifically but not due to its binding of Ca2+ in the medium .


Variation in reproductive traits in European passerines in relation to nesting site: allometric scaling to body weight or adaptive variation?
The relationships between different reproductive traits and body weight, in relation to nesting site, were studied in European passerines. Body weight explains a significant proportion of the variance in clutch size, egg weight, clutch mass, and incubation period. Whereas clutch size decreases, the other three traits increase with body weight. Even though both clutch mass and clutch size are related to body size, for a given body weight, concealed nesters lay a larger clutch and a greater clutch mass than open-nesters.


Development and Function of Effector Regulatory T Cells.
Distinguishing self from nonself is a unique feature of the immune system. Although most self-reactive T cells are eliminated in the thymus, a few rogue cells escape the negative selection process and have the potential to mediate autoimmune disease. Over the last decade, there has been a vast improvement in our understanding of the cellular mechanisms that evolved to dampen the deleterious effects of these self-reactive T cells. In particular, T cells expressing the transcription factor FoxP3, known as regulatory T (Treg) cells, play a central role in maintaining immune homeostasis and suppressing autoimmune responses. In addition, Treg cells are endowed with the ability to suppress diverse inflammatory responses both in lymphoid and in nonlymphoid tissues. This requires Treg cells to undergo a peripheral differentiation and specialization program that results in the emergence of effector Treg (eTreg) cells that are characterized by their ability to produce high amounts of immunosuppressive molecules, including IL-10. This chapter discusses the recent advances in our understanding of the mechanisms governing the differentiation, migration, and maintenance of eTreg cells, in particular in nonlymphoid tissues, in health and disease.


Chemically and mechanically isolated nanocellulose and their self-assembled structures.
Cellulose nanocrystals (CNCs) and nanofibrils (CNFs) have been isolated from pure rice straw cellulose via sulfuric acid hydrolysis, mechanical blending and TEMPO-mediated oxidation to 16.9%, 12% and 19.7% yields, respectively. Sulfuric acid hydrolysis produced highly crystalline (up to 90.7% CrI) rod-like (3.96-6.74 nm wide, 116.6-166 nm long) CNCs with similarly negative surface charges (-67 to -57 mV) and sulfate contents but decreasing yields and dimensions with longer hydrolysis time. Mechanical defibrillated CNFs were 82.5% crystalline and bimodally distributed in sizes (2.7 nm wide and 100-200 nm long; 8.5 nm wide and micrometers long). TEMPO mediated oxidation liberated the most uniform, finest (1.7 nm) and micrometer long, but least crystalline (64.4% CrI) CNFs. These nanocellulose self-assembled into submicron (153-440 nm wide) fibers of highly crystalline (up to 90.9% CrI) cellulose Iβ structure upon rapid freezing (-196 °C) and freeze-drying. The self-assembling behaviors were analyzed based on nanocellulose dimensions, specific surfaces and surface chemistries.


Characterization of the paraoxonase system in follicular fluid of women subjected to an ovarian stimulation cycle.
Reactive oxygen species (ROS) and antioxidants are involved in the regulation of reproductive processes. Previous studies in infertile women undergoing an ovarian stimulation cycle have suggested a possible role for ROS in the occurrence of conception in In Vitro Fertilization. In this context the control of the redox balance of follicular fluid becomes essential for reproduction, so that the presence of enzymes with antioxidant activities, such as the paraoxonase (PON) system, would play a role in maintaining this balance. The objective of this work was a) to characterize the paraoxonase system in follicular fluid of women undergoing a controlled ovarian stimulation cycle, analysing the associated PON1, PON2, and PON3 activities, and b) to study the possible involvement of the PON system in follicular maturation. The enzyme activities were quantified in follicular fluid from large and small follicles from women undergoing an ovarian stimulation cycle in the IVI-Bilbao clinic. PON activities were quantified using spectrophotometric and HPLC techniques. Statistical comparisons were performed using the Student's t-test for paired data. Results indicate that follicular fluid presents paraoxonase activities which are detectable by the methods developed in this study. PON activities were associated with follicular maturation, suggesting that the PON system plays a role in oocyte maturation. This work was supported by research grants from the Ministry of Health and Consumption (FIS/FEDER PI11/02559), the Basque Country Government (Dep. Education, Universities and Research ref. IT687-13, and DCIT ref. S-PE13UN063), and UPV/EHU (CLUMBER UFI11/20 and PES13/58). The work was approved by the Ethics Committee of the UPV/EHU (CEISH/96/2011/RUIZLARREA), and performed according to the UPV/EHU and IVI-Bilbao agreement (Ref. 2012/01).


Dynamics of Metabolic Pathways and Stress Response Patterns during Human Neural Stem Cell Proliferation and Differentiation.
Understanding early nervous system stress response mechanisms is crucial for studying developmental neurotoxicity and devising neuroprotective treatments. We used hiPSC-derived long-term self-renewing neuroepithelial stem (lt-NES) cells differentiated for up to 12 weeks as an in vitro model of human neural development. Following a transcriptome analysis to identify pathway alterations, we induced acute oxidative stress (OS) using tert-butyl hydroperoxide (TBHP) and assessed cell viability at different stages of neural differentiation. We studied NRF2 activation, autophagy, and proteasomal function to explore the contribution and interplay of these pathways in the acute stress response. With increasing differentiation, lt-NES cells showed changes in the expression of metabolic pathway-associated genes with engagement of the pentose phosphate pathway after 6 weeks, this was accompanied by a decreased susceptibility to TBHP-induced stress. Microarray analysis revealed upregulation of target genes of the antioxidant response KEAP1-NRF2-ARE pathway after 6 weeks of differentiation. Pharmacological inhibition of NRF2 confirmed its vital role in the increased resistance to stress. While autophagy was upregulated alongside differentiation, it was not further increased upon oxidative stress and had no effect on stress-induced cell loss and the activation of NRF2 downstream genes. In contrast, proteasome inhibition led to the aggravation of the stress response resulting in decreased cell viability, derangement of NRF2 and KEAP1 protein levels, and lacking NRF2-pathway activation. Our data provide detailed insight into the dynamic regulation and interaction of pathways involved in modulating stress responses across defined time points of neural differentiation.


Absorbable gelatin plug does not prevent iatrogenic preterm premature rupture of membranes after fetoscopic laser surgery for twin-twin transfusion syndrome.
Despite fetoscopic laser ablation (FLA) having emerged as an effective treatment for twin-twin transfusion syndrome (TTTS), major postintervention challenges, such as iatrogenic preterm premature rupture of membranes (iPPROM), remain. A chorioamniotic plug (CAP) made of absorbable gelatin sponge has been used to seal the trocar entry site in the chorioamniotic layers to promote healing and prevent iPPROM, yet the results have been equivocal. Our objective was to test the hypothesis that, following FLA for TTTS, iPPROM may be prevented by placement of an absorbable gelatin CAP. A retrospective cohort study was performed on prospectively collected data from 134 consecutive patients who underwent FLA for TTTS. The decision for CAP placement was at the discretion of the physician. Preoperative, operative and postoperative variables were collected and analyzed. The primary outcome was the incidence of iPPROM, and the secondary outcomes were procedure-to-delivery interval and gestational age at delivery. Comparative statistical analysis was performed as appropriate. A CAP was placed in 72 (54%) of 134 patients. Factors significantly associated with CAP placement were higher gravidity (P = 0.03), anterior placentation (P = 0.04), general endotracheal intubation (P = 0.02) and a cannula diameter of 12-Fr (P = 0.003). There were no differences between CAP and no-CAP groups in the rate of iPPROM (39% vs 34%, respectively; P = 0.42) or in the procedure-to-delivery interval (65.3 ± 34.7 days vs 58.2 ± 30.8 days, respectively; P = 0.21). The gestational age at delivery was later in the CAP group compared with the no-CAP group (30.7 ± 4.5 weeks vs 28.9 ± 3.9 weeks, respectively; P = 0.02). CAP did not reduce the overall risk for iPPROM and did not increase the procedure-to-delivery interval. Further research is needed to identify other methods to prevent iPPROM and prolong pregnancy after laser therapy.


THOC2 and THOC5 Regulate Stemness and Radioresistance in Triple-Negative Breast Cancer.
Triple-negative breast cancer (TNBC) is the most aggressive subtype of breast cancer. Radioresistance and stemness are substantial obstacles to TNBC treatment. The THO complex (THOC) is a subunit of the TRanscription-EXport complex that functions in the coupling of transcription to nascent RNA splicing, elongation, and export. However, its role in regulating TNBC therapeutic resistance is not reported yet. In this study, the authors demonstrate that cancer stem cells are enriched in radioresistant TNBC cells and describe the role of the THOC in regulating TNBC radioresistance and stemness. The authors find that THOC2 and THOC5 are upregulated in radioresistant TNBC cells and associated with a poor prognosis in TNBC patients. Further investigation reveals that THOC2 promotes the stem-like properties and radioresistance of TNBC cells in a THOC5-dependent manner by facilitating the release of sex-determining region Y (SRY)-box transcription factor 2 (SOX2) and homeobox transcription factor (NANOG) transcripts from the nucleus. Silencing THOC2 or THOC5 expression decreases the protein expression of SOX2 and NANOG, depletes the stem-like properties, and causes radiosensitization in these TNBC cells. Moreover, THOC2 or THOC5 depletion blocks the xenograft tumorigenesis and growth of radioresistant TNBC in vivo. These findings uncover the novel correlations of THOC with TNBC stemness and therapeutic resistance, proposing alternative therapeutic strategies against relapsed TNBC.


Novel generation of hormone receptor specificity by amino terminal processing of peptide YY.
The physiological significance of multiple Y receptors has not been determined since until recently only one form of endogenous agonists was known, namely PYY1-36 and NPY1-36. Recently, a new molecular form of PYY was characterized as des(Tyr-Pro)PYY (PYY3-36 or PYY-II). Its ability to interact at various Y receptors was not characterized. Analytical chromatography of fresh canine colon extracts shows two peaks of immunoreactivity eluting in the positions of PYY-II and PYY1-36 (PYY). PYY-II was about 40% of the total PYY immunoreactivity indicating that it is one of the major forms of PYY expressing its biological activity. It is shown that PYY-II will not displace label from the Y1 receptors found on a human neuroblastoma cell line. It is further shown that PYY-II is as potent as PYY for the inhibition of pancreatic secretion, which must occur through Y2 receptors. The enzymatic removal of Tyr-Pro from PYY to form PYY-II must therefore regulate the relative expression of a non-selective agonist (PYY) to a highly selective Y2 agonist (PYY-II). Amino terminal processing of PYY represents a novel type of regulation of peptide hormone specificity. It has important biological implications for PYY and potential relevance for other peptide hormone receptor systems.


Combined tubal and multiple intrauterine pregnancies following ovulation induction.
An infertile woman who conceived after human menopasual gonadotropin-induced ovulation experienced ovarian hyperstimulation and a quintuplet gestation--an ectopic and a quadruplet intrauterine pregnancy. The intrauterine pregnancy was confirmed by B-scan ultrasonography about the time that the tubal pregnancy was ruptured. Salpingectomy did not interfere with the intrauterine pregnancy, but in spite of a cervical cerclage the patient developed premature labor after about 26 weeks of gestation.


Expression of nm23 in gastric carcinoma: association with tumor progression and poor prognosis.
Expression of nm23 has been shown to be inversely correlated with the metastatic potential of several human cancers. In the current study, the expression and prognostic impact of nm23 was immunohistochemically studied in 413 curatively resected gastric carcinomas. Tumor sections of the 413 gastric carcinomas were stained with a polyclonal antibody that was raised against the nm23-H1/NDP kinase A, which is identical to the nm23-H1 gene product. Expression of nm23 was detected in 84.5% (n = 349) of all tumors, in the majority of cases (71.2%) causing a homogeneous staining reaction in more than 75% of tumor cells. Expression of nm23 was positively correlated with the intestinal type of tumor, according to the Lauren classification and advanced pT categories, and was also correlated with the presence of blood and lymphatic vessel invasion. In contrast, no correlation could be demonstrated between nm23 expression and lymph node involvement. As shown in univariate analysis, patients with nm23 positive tumors, especially those with nm23 positive diffuse-type carcinomas, had significantly shorter overall survival than patients with nm23 negative tumors (P = 0.03 and P = 0.0065, respectively). However, in a multivariate analysis that included the prognostic parameters pT category, pN category, and blood and lymphatic vessel invasion, this prognostic impact was not maintained. In contrast to results for breast and colorectal carcinomas, our results for 413 gastric carcinomas showed that expression of the designated metastasis suppressor gene nm23 is correlated with aggressive tumor growth and poor prognosis but is not an independent prognostic marker.


Occurrence, distribution, and bioaccumulation of antibiotics in coastal environment of Dalian, China.
Seawater, sediment, and aquatic organism samples were collected from 20 sampling sites in coastal environment of Dalian in August, 2011. The occurrence, distribution, and bioaccumulation of 20 antibiotics categorizing into three groups, including 14 sulfonamides (SAs), two chloramphenicols (CAPs) and four tetracyclines (TCs), were investigated. The results suggested that tetracyclines were the predominant antibiotics in the seawater (range: 2.11-9.23 ng L(-1)), while sulfonamides were the dominant antibiotics in both sediments (range: 1.42-71.32 μg kg(-1)) and aquatic organisms (range: 2.18-63.87 μg kg(-1)). The sorption coefficient Kd,s values revealed that sulfameter, sulfadiazine, sulfamethoxypyridazine, sulfamonomethoxine, chloramphenicol, and doxycycline presented higher sorption capacities than the other antibiotics. The average BAFs suggested that sulfamethazine, sulfamethiazole, sulfamonomethoxine, and doxycycline were potentially bioaccumulative, while sulfadiazine, sulfameter, sulfamethoxypyridazine, and chloramphenicol were bioaccumulative.


Effects of autophagy inhibition by chloroquine on hepatic stellate cell activation in CCl4-induced acute liver injury mouse model.
Hepatic stellate cells (HSCs) activation, a critical event in liver fibrosis, has been recently shown to be related to autophagy. Determine whether chloroquine (CQ) could affect (i) the activation of HSC in vivo and (ii) the hepatic damage in a mice acute liver injury model. The acute liver injury was induced in BALB/c mice by carbon tetrachloride (CCl4 group); 24 h before and after CCl4 administration animals were treated by CQ (CCl4 + CQ group). As control, mice treated by olive oil were considered. After 48 h from CCl4 /olive oil administration, blood samples, liver tissues, and HSCs were harvested for analysis. In vivo, CQ attenuates CCl4 -induced acute liver damage as evidenced by (i) the reduction of liver enlargement, (ii) the reduction of liver swelling and necrosis also supported by a certain decrease of circulating transaminases level, and (iii) the reduction of liver fibrosis evaluated by collagen deposition and α-sma protein expression. In HSCs isolated from CQ treated group, we observed the inhibition of autophagy proved by the increase in p62 protein and the decrease of lc3 protein. In addition, CQ reduced the expression of the HSCs activation markers α-sma/collagen-I and down-regulated the expression of the proliferative marker ki67. The autophagy attenuation exerted by CQ together with the reduction of the expression of the proliferation marker in HSCs can lessen the acute liver damage potentially opening the way to novel therapeutic approaches for hepatic fibrosis.


Treatment of refractory atopic dermatitis using 'wet-wrap' dressings and diluted corticosteroids: results of standardized treatment in both children and adults.
'Wet-wrap' dressings with diluted corticosteroids form an alternative treatment in patients with refractory atopic dermatitis (AD). To evaluate a standardized treatment, using wet-wrap dressings with diluted corticosteroids, in patients with refractory AD. Results of treatment, complications and possible side effects were retrospectively evaluated in 14 children and 12 adults. Skin lesions improved dramatically during 1 week of inpatient treatment. A significant decrease in early-morning serum cortisol levels was measured. Levels below the normal range were only observed after 1 week in 2 adults and on day 4 in 3 children. Suppression of the hypothalamus-pituitary-adrenal-cortex axis in 1 adult and a new exacerbation of AD in 2 children and 3 adults complicated long-term treatment at home. Additional complications included folliculitis, a Pseudomonas aeruginosa infection, a secondary bacterial infection and refractory skin lesions between bandages. Wet-wrap dressings and diluted corticosteroids form an effective treatment in patients with refractory AD.


An adaptive correlation ratemeter: a new method for Doppler fetal heart rate measurements.
For use in a cardiotocograph, ultrasound Doppler fetal singles have been thought to provide merely a limited reliability of the fetal heart rate record because of the difficulty of signal processing to obtain a consistent trigger, although the signal itself is easy to obtain with an excellent signal-to-noise ratio. However, the authors have developed a unique signal processing system using a correlation technique with an automatic adaptation algorithm, which solved almost all of the difficulties associated with the old Doppler cardiotocograph and showed that the ultrasound Doppler fetal signal can also provide heart rate records compatible to that obtained by a fetal ECG system. The authors are sure that their method is a most promising one in the coming age of microprocessor-oriented instrumentation.


Inhibition of growth and ochratoxin A biosynthesis in Aspergillus carbonarius by flavonoid and nonflavonoid compounds.
The effect of naturally occurring phenolic compounds on Aspergillus carbonarius growth and ochratoxin A (OTA) production was studied. Caffeic acid and the flavonoids, rutin and quercetin, were added to Czapek Yeast Extract agar at concentrations ranging between 50 and 500 mg/l. All phenolic compounds had a significant influence on growth rate and lag phase of A. carbonarius at 250 mg/l. The growth was completely inhibited with 500 mg/l. In comparison with the control, a significant decrease in OTA production was observed with all phenolic compounds. In general, effect on growth was less evident than effect on toxin production. An inhibitory effect on growth and OTA production, as concentration was increased was observed in all cases. The response of A. carbonarius to the flavonoids, rutin and quercetin, was similar. The inhibitory effect of these natural phenolic compounds on fungal growth and OTA production could be an alternative to the use of chemical fungicides.


[Epidemiology and Resistance Mechanisms of Group B Streptococci in Late-pregnant Maternal Birth Canal].
To study the antibiotic-resistant rate of group B streptococci (GBS) in obstetric canal of late-pregnant women, evaluate the antibiotic-resistant status and finally to support the GBS prevention and curing by proper antibiotics. 31 pregnant women between 35 to 37 gestational weeks were included, for whom the antibiotic sensitivity as well as the drug (erythromycin and clindamycin) resistance genes of GBS in obstetric canal was analyzed. 12 (38. 7%) strains of GBS were resistant to clindamycin, while 21 (67. 7%) to erythromycin, within which 12 strains were intrinsic phenotype - cMLS type-clindamycin resistance, other 9 were active efflux phenotype - MS type-clindamycin sensitive and all of which were confirmed by Double disk diffusion method. Eleven strains were mef (A) positive, and 12 strains were erm (B) positive, in which 3 with erm (C). In our research the GBS strains show a high erythromycin and clindamycin resistance rate. The resistance of our GBS strains are mainly caused by the ribosomal target changes induced by erm (B) and the increased efflux of clindamycin induced by mef (A).


Effect of RNA and protein synthesis inhibitors on the release of inflammatory mediators by macrophages responding to phorbol myristate acetate.
The interaction of phorbol myristate acetate with resident populations of mouse peritoneal macrophages causes an increased release of arachidonic acid followed by increased synthesis and secretion of prostaglandin E2 and 6-keto-prostaglandin F1 alpha. In addition, phorbol myristate acetate causes the selective release of lysosomal acid hydrolases from resident and elicited macrophages. These effects of phorbol myristate acetate on macrophages do not cause lactate dehydrogenase to leak into the culture media. The phorbol myristate acetate-induced release of arachidonic acid and increased synthesis and secretion of prostaglandins by macrophages can be inhibited by RNA and protein synthesis inhibitors, whereas the release of lysosomal hydrolases is unaffected. 0.1 microgram/ml actinomycin D blocked the increased prostaglandin production due to this inflammatory agent by more than 80%, and 3 microgram/ml cycloheximide blocked prostaglandin production by 78%. Similar results with these metabolic inhibitors were found with another stimulator of prostaglandin production, zymosan. However, these inhibitors do not interfere with lysosomal hydrolase releases caused by zymosan or phorbol myristate acetate. It appears that one of the results of the interaction of macrophages with inflammatory stimuli is the synthesis of a rapidly turning-over protein which regulates the production of prostaglandins. It is also clear that the secretion of prostaglandins and lysosomal hydrolases are independently regulated.


Age Moderates Perceived COVID-19 Disruption on Well-Being.
It is not fully understood how large-scale events affect well-being. Older adults showed the highest levels of resilience following the September 11th (9/11) terrorist attacks, but during the severe acute respiratory syndrome outbreak, there were no age-related differences in well-being. The current study examined the Coronavirus Disease 2019 (COVID-19) disruption on well-being throughout adulthood. Perceived stress and affect were examined in 166 community-dwelling adults (Mage = 35.65; SD = 15.53; range = 18-79) in relation to the perceived disruption of the COVID-19 pandemic to their lives. A significant moderation was found for age and COVID-19 disruption on perceived stress [F(5, 153) = 8.88, p < .05, R2 = .22] and negative affect [F(5, 154) = 4.91, p < .05, R2 = .14], but not for positive affect. For participants over 50, those who rated COVID-19 as a low or high disruption had similar scores on stress and negative affect, but with younger aged participants, perceiving high disruption corresponded with higher levels of stress and negative affect. Findings are consistent with the strength and vulnerability integration (SAVI) model, wherein older adults try to maintain positive emotional well-being, with middle-aged and older adults in the current study having experienced less negative impact on well-being. Middle-aged and older adults may be better able to regulate negative emotions from COVID-19 than younger adults. SAVI proposes a greater negative impact on older adults when they experience sustained stressors; as the challenges with COVID-19 continue, further data will need to be examined.


Ichthyosiform eruption associated with lamivudine in a patient with chronic hepatitis-B infection.
The side-effects of antiretroviral agents have been widely reported in the literature. Lamivudine is an antiretroviral agent, and skin eruption because of this agent has been rarely reported. Previous reports regarding these few side-effects of lamivudine include angioedema, urticaria, anaflactoid reactions, allergic contact dermatitis and alopecia. There is no report of an ichthyosiform eruption associated with lamivudine. The authors present a case of 43-year-old female patient presenting with an ichthyosiform eruption during lamivudine therapy for chronic hepatitis-B for the first time.


MyD88 signaling is not essential for induction of antigen-specific B cell responses but is indispensable for protection against Streptococcus pneumoniae infection following oral vaccination with attenuated Salmonella expressing PspA antigen.
TLRs directly induce innate host defense responses, but the mechanisms of TLR-mediated adaptive immunity remain subject to debate. In this study, we clarified a role of TLR-mediated innate immunity for induction of adaptive immunity by oral vaccination with a live recombinant attenuated Salmonella enteric serovar Typhimurium vaccine (RASV) strain expressing Streptococcus pneumoniae surface protein A (PspA) Ag. Of note, oral or intranasal vaccination with RASV expressing PspA resulted in identical or even significantly higher levels of PspA-specific IgG and IgA responses in the systemic and mucosal compartments of MyD88(-/-) mice of either BALB/c or C57BL/6 background when compared with those of wild-type mice. Although PspA-specific CD4(+) T cell proliferation in the MyD88(-/-) mice was minimal, depletion of CD4(+) T cells abolished PspA-specific IgG and IgA responses in the MyD88(-/-) mice of BALB/c background. Of the greatest interest, MyD88(-/-) mice that possessed high levels of PspA-specific IgG and IgA responses but minimal levels of CD4(+) T cell responses died earlier than nonvaccinated and vaccinated wild-type mice following i.v. or intranasal challenge with virulent S. pneumoniae. Taken together, these results suggest that innate immunity activated by MyD88 signals might not be necessary for Ag-specific Ab induction in both systemic and mucosal sites but is critical for protection following oral vaccination with attenuated Salmonella expressing PspA.


MiR-708 promotes steroid-induced osteonecrosis of femoral head, suppresses osteogenic differentiation by targeting SMAD3.
Steroid-induced osteonecrosis of femoral head (ONFH) is a serious complication of glucocorticoid (GC) use. We investigated the differential expression of miRs in the mesenchymal stem cells (MSCs) of patients with ONFH, and aimed to explain the relationship between GC use and the development of MSC dysfunction in ONFH. Cells were collected from bone marrow of patients with ONFH. Samples were assigned to either GCs Group or Control Group at 1:1 matched with control. We then used miRNA microarray analysis and real-time PCR to identify the differentially expressed miRs. We also induced normal MSCs with GCs to verify the differential expression above. Subsequently, we selected some of the miRs for further studies, including miRNA target and pathway prediction, and functional analysis. We discovered that miR-708 was upregulated in ONFH patients and GC-treated MSCs. SMAD3 was identified as a direct target gene of miR-708, and functional analysis demonstrated that miR-708 could markedly suppress osteogenic differentiation and adipogenesis differentiation of MSCs. Inhibition of miR-708 rescued the suppressive effect of GC on osteonecrosis. Therefore, we determined that GC use resulted in overexpression of miR-708 in MSCs, and thus, targeting miR-708 may serve as a novel therapeutic biomarker for the prevention and treatment of ONFH.


205Tl+ as a spectroscopic probe of the monovalent cation binding sites of bovine plasma activated protein C and des-1-41-light-chain-activated protein C.
The kinetic properties of the stimulation by Tl+ of the amidase activity of bovine plasma activated protein C (APC) and a limited-proteolytic derivative of this enzyme, des-1-41-light chain APC (GDAPC), which has no remaining gamma-carboxyglutamic acid residues, have been compared, along with a 205Tl+ NMR analysis of the interaction of this cation with these enzymes, at 6 degrees C. In contrast to other monovalent cations, the productive kinetic complex of Tl+ and APC involves only a single Tl+ site, or class of sites, and is similar to GDAPC in this regard. In the case of each enzyme, the kinetic mechanism that best describes the participation of Tl+ is a rapid equilibrium type with random addition of the cation and substrate to the enzyme. The dissociation constants of the Tl+ X APC and Tl+ X GDAPC complexes have been determined by NMR analysis and have been found to be very similar to the same constants as calculated by kinetic means. These cation sites are also present intact on each zymogen, demonstrating that they are not generated as a result of activation. Our results also show that the Ca2+ binding sites of these proteins are exclusive of the T1+ site and that some interference with Tl+ binding is exercised by an active site-directed affinity label. We conclude that Tl+ can be effectively employed as a spectroscopic probe of the monovalent cation sites that serve an extensive stimulatory role in the amidolytic and esterolytic activities of APC.


Daily prostate targeting using implanted radiopaque markers.
A system has been implemented for daily localization of the prostate through radiographic localization of implanted markers. This report summarizes an initial trial to establish the accuracy of patient setup via this system. Before radiotherapy, three radiopaque markers are implanted in the prostate periphery. Reference positions are established from CT data. Before treatment, orthogonal radiographs are acquired. Projected marker positions are extracted semiautomatically from the radiographs and aligned to the reference positions. Computer-controlled couch adjustment is performed, followed by acquisition of a second pair of radiographs to verify prostate position. Ten patients (6 prone, 4 supine) participated in a trial of daily positioning. Three hundred seventy-four fractions were treated using this system. Treatment times were on the order of 30 minutes. Initial prostate position errors (sigma) ranged from 3.1 to 5.8 mm left-right, 4.0 to 10.1 mm anterior-posterior, and 2.6 to 9.0 mm inferior-superior in prone patients. Initial position was more reproducible in supine patients, with errors of 2.8 to 5.0 mm left-right, 1.9 to 3.0 mm anterior-posterior, and 2.6 to 5.3 mm inferior-superior. After prostate localization and adjustment, the position errors were reduced to 1.3 to 3.5 mm left-right, 1.7 to 4.2 mm anterior-posterior, and 1.6 to 4.0 mm inferior-superior in prone patients, and 1.2 to 1.8 mm left-right, 0.9 to 1.8 mm anterior-posterior, and 0.8 to 1.5 mm inferior-superior in supine patients. Daily targeting of the prostate has been shown to be technically feasible. The implemented system provides the ability to significantly reduce treatment margins for most patients with cancer confined to the prostate. The differences in final position accuracy between prone and supine patients suggest variations in intratreatment prostate movement related to mechanisms of patient positioning.


In-Air and Underwater Hearing in the Great Cormorant (Phalacrocorax carbo sinensis).
Hearing thresholds of a great cormorant (Phalacrocorax carbo) were measured in air and under water using psychophysics. The lowest thresholds were at 2 kHz (45 dB re 20 μPa root-mean-square [rms] in air and 79 dB re 1 μPa rms in water). Auditory brainstem response measurements on one anesthetized bird in air indicated an audiogram with a shape that resembled the one achieved by psychophysics. This study suggests that cormorants have rather poor in-air hearing abilities compared with other similar-size birds. The hearing capabilities in water are better than what would have been expected for a purely in-air adapted ear.


Mathematical modeling of enzyme production using Trichoderma harzianum P49P11 and sugarcane bagasse as carbon source.
A mathematical model to describe the kinetics of enzyme production by the filamentous fungus Trichoderma harzianum P49P11 was developed using a low cost substrate as main carbon source (pretreated sugarcane bagasse). The model describes the cell growth, variation of substrate concentration and production of three kinds of enzymes (cellulases, beta-glucosidase and xylanase) in different sugarcane bagasse concentrations (5; 10; 20; 30; 40 gL(-1)). The 10 gL(-1) concentration was used to validate the model and the other to parameter estimation. The model for enzyme production has terms implicitly representing induction and repression. Substrate variation was represented by a simple degradation rate. The models seem to represent well the kinetics with a good fit for the majority of the assays. Validation results indicate that the models are adequate to represent the kinetics for a biotechnological process.


[ZRS mutations in two Chinese Han families featuring triphalangeal thumbs and preaxial polydactyly].
To identify the causative mutations in two Chinese Han families featuring triphalangeal thumbs (TPT) and preaxial polydactyly (PPD). Blood samples were collected from 9 members (2 affected) from family 1 and 14 members (7 affected) from family 2. After genomic DNA was extracted, the ZPA regulatory sequence (ZRS) region was analyzed with real-time quantitative PCR (qPCR) and Sanger sequencing. For family 1, haplotypes compassing the ZRS were also analyzed with short tandem repeats (STR) and single nucleotide changes. No copy number mutation around the ZRS region was found in both families. Two heterogeneous mutations in the ZRS (406A>G and 105C>G) were found to co-segregate with the TPT/PPD malformation in family 1 and 2, respectively. Neither mutation was detected in 200 healthy individuals. Haplotype analysis and Sanger sequencing of family 1 indicated that the first TPT/PPD patient in the family was both germline and somatic mosaic for the 406A>G mutation. Two pathogenic ZRS mutations, 105C>G and 406A>G, have been identified in two Chinese Han families with TPT/PPD, among which the 406A>G mutation was de novo.


Change in Alcohol Use and Association with Positive and Negative Emotions: Results from an Alcohol Treatment Study with Hepatitis C Patients.
Few studies exist on the change over time in positive and negative emotions during treatment for alcoholism disorders. We aimed to evaluate relationship between alcohol reduction and change in positive and negative emotions. Chronic HCV patients (n=174) with alcohol use received brief alcohol counseling. Participants completed the PANAS-Short Form, MHC-Short Form, and the Alcohol Timeline Follow back at baseline and 3, 6, and 12 months. Decreases in alcohol use were related to decreased negative emotions from baseline to 3 months, baseline to 6 months and baseline to 12 months. Decreases in alcohol use were associated with increased positive emotions from baseline to 12 months but not sooner.