Morphology, anatomy and histochemistry of Salicornioideae (Chenopodiaceae) fruits and seeds.
The subfamily Salicornioideae (Chenopodiaceae) are a taxonomically difficult group largely due to the lack of diagnostic characters available to delineate tribal- and generic-level boundaries; a consequence of their reduced floral and vegetative features. This study examined the variation in fruits and seeds across both tribes of the Salicornioideae to assess if characters support traditional taxonomic sections. Light microscopy, environmental scanning electron microscopy and anatomical ultra-thin sectioning were employed to examine variation in fruits and seeds. Sixty-eight representatives across 14 of the 15 genera currently recognized within the tribes Halopeplideae and Salicornieae were examined to determine whether characters support current taxonomic groups. Characters such as seed coat structure, embryo shape, seed orientation, the forms of seed storage proteins and carbohydrates show variation within the Salicornioideae and may be phylogenetically useful. The campylotropous ovule typical of the Chenopodiaceae generally results in a curved embryo; however, many Halosarcia and Sclerostegia species have straight embryos and in Salicornia and Sarcocornia the large peripheral embryo appears bent rather than curved. Seed coat ornamentation of Microcnemum and Arthrocnemum is distinct from other Salicornioideae as the elongated epidermal cells of the exotesta have convex walls. Histochemical stains of anatomical sections of cotyledon cells showed protein bodies were variable in shape, and starch grains were present in some species, namely Salicornia bigelovii, S. europaea and Allenrolfea occidentalis. While fruits and seeds were found to be variable within the subfamily, no synapomorphic characters support the tribe Halopeplideae as these genera have crustaceous seed coats, curved embryos and abundant perisperm; features characteristic of many of the tribe Salicornieae. The endemic Australian genera are closely related and few seed and fruit characters are diagnostic at the generic level. Nineteen characters identified as being potentially informative will be included in future phylogenetic analyses of the subfamily.


KAT5 acetylates cGAS to promote innate immune response to DNA virus.
The DNA sensor cGMP-AMP synthase (cGAS) senses cytosolic microbial or self DNA to initiate a MITA/STING-dependent innate immune response. cGAS is regulated by various posttranslational modifications at its C-terminal catalytic domain. Whether and how its N-terminal unstructured domain is regulated by posttranslational modifications remain unknown. We identified the acetyltransferase KAT5 as a positive regulator of cGAS-mediated innate immune signaling. Overexpression of KAT5 potentiated viral-DNA-triggered transcription of downstream antiviral genes, whereas a KAT5 deficiency had the opposite effects. Mice with inactivated Kat5 exhibited lower levels of serum cytokines in response to DNA virus infection, higher viral titers in the brains, and more susceptibility to DNA-virus-induced death. Mechanistically, KAT5 catalyzed acetylation of cGAS at multiple lysine residues in its N-terminal domain, which promoted its DNA-binding ability. Our findings suggest that KAT5-mediated cGAS acetylation at its N terminus is important for efficient innate immune response to DNA virus.


Massive Post-Partum Haemorrhage In A SARS-Cov-2 Positive Primi Gravid Patient: A Case Report.
A 20-year-old, primi gravid, south Asian woman, was referred from secondary care to Aga Khan University Hospital for further management of primary post-partum haemorrhage after a normal vaginal delivery, on July 12, 2021. She was otherwise healthy, with no known comorbidity. She tested positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during her admission, on account of universal Covid-19 testing policy. Initially, conservative measures were instituted to control bleeding, including bilateral internal iliac artery ligation, concluding with hysterectomy to stop the haemorrhage. Multiple units of blood products had to be transfused. Obstetrical haemorrhage being the menace in this case, the possibility of Covid-19 positive obstetrics patients being more susceptible to massive post-partum haemorrhage, requiring more radical life-saving procedures to be conducted earlier, was considered. In future, more observational studies will be needed to determine any association between Covid-19 infection and obstetric haemorrhage.


Cortical and subcortical neurodegeneration is associated with HIV neurocognitive impairment.
To determine the association of markers of regional neurodegeneration (ND) at autopsy to degree of neurocognitive impairment in persons with HIV. In a prospectively followed cohort of HIV-infected individuals we examined the relationship between antemortem neuropsychological (NP) abilities and postmortem neuropathological data. Twenty-seven HIV-infected individuals with both neuropsychological and neuropathological data were identified. Laser confocal scanning microscopy was used to determine the degree of ND based on: (1) microtubule-associated protein (MAP2; reflecting neuronal cell bodies and dendrites) and (2) synaptophysin (SYN; a measure of presynaptic terminals). A regional combined score, based on the distribution of percentage neuropil occupied by MAP2 and SYN and emphasizing severity of ND, was created for each brain region: midfrontal cortex, hippocampus, and putamen. The regional combined scores from each brain region studied were better correlated with level of global NP impairment than measures of SYN and MAP2 individually. In a regression, hippocampal and putamen regional combined scores were independent predictors of degree of antemortem NP impairment (F(3,23) = 6.17; P < 0.01; R2 = 0.45). The correlations among regional ND measures demonstrated that ND is unevenly distributed across multiple brain regions. As the anatomic distribution and temporal progression of neuropathologic changes appears to differ across individuals, it is important to consider both cortical and subcortical brain regions in studies of neuropathogenesis and treatment of HIV-related brain disease. Furthermore, combining information from several markers of neural injury provided the strongest association with degree of neurocognitive impairment during life.


Inflammatory cells and chronic obstructive pulmonary disease.
A major contributory factor to the development of chronic obstructive pulmonary disease (COPD) is the inflammatory response to cigarette smoke. However, when those with COPD stop smoking, a continuous cycle of inflammation can lead to continued decline in lung function. Understanding the role of inflammatory cells in COPD is difficult because it is a mixture of diseases--bronchitis, small airways disease and emphysema--that exhibit different patterns of inflammation and different pathology. Neutrophils and macrophages have been implicated in this process; they release proteolytic enzymes and generate oxidants, which cause tissue damage, as well as cytokines and chemokines, which can potentiate inflammation and trigger an immune response. Analysis of sputum and bronchoalveolar lavage fluid shows increases in both neutrophils and macrophages in respiratory secretions in COPD subjects; neutrophils are the predominant cell in the conducting airways, whereas macrophages are the major cell in secretions from the small airways and parenchyma. Airway tissue neutrophils are increased in the large and small airways during infection and exacerbations, whilst parenchymal neutrophil numbers are inversely related to alveolar wall destruction, suggesting that they are not involved in the progression of emphysema. Macrophages are increased throughout the respiratory tract airway lumen and epithelium in COPD and are positively related to severity of disease, airway obstruction and degree of alveolar wall damage in emphysema. Unactivated T-lymphocytes do not linger in lung tissue. Activated (eg due to antigenic stimulus) memory T cells home in to the lung and act as effector cells. CD-8+ T cell differentiation into memory cells is facilitated by CD4+ T cells. Binding of CD-8+ T cells to collagen stimulates proliferation and mediator production which may contribute to the inflammatory response. CD8+ cytotoxic/suppressor T cells release cytotoxic perforins and granzyme B which cause cell death and apoptosis, a feature of emphysema. Lung secretions contain only a small percentage of T cells; most T-lymphocytes reside in the subepithelial and smooth muscle region of the tissue. During COPD, there is either an increase in the CD8+/CD4+ ratio of T cells, or an increase in the in total numbers of both CD8+ and CD4+ T cells, in the tissue. Smoking status, smoking history, degree of airway obstruction and emphysema are all related to increased CD8+ cells and/or CD8+/CD4+ ratio. During severe emphysema requiring lung volume reduction surgery, there is a considerable increase in macrophages, neutrophils, eosinophils, CD4+ and CD8+ T cells which relates to the severity of the disease. Interestingly, the marked increase in luminal CD8+ cells results in an increased ratio of CD8+/CD4+ T cells that is not seen in the parenchymal tissue. The florid inflammation observed in severe emphysema is suggested to be related to latent viral infection.


Reconstructing phylogenetic relationships based on repeat sequence similarities.
A recent phylogenetic method based on genome-wide abundance of different repeat types proved to be useful in reconstructing the evolutionary history of several plant and animal groups. Here, we demonstrate that an alternative information source from the repeatome can also be employed to infer phylogenetic relationships among taxa. Specifically, this novel approach makes use of the repeat sequence similarity matrices obtained from the comparative clustering analyses of RepeatExplorer 2, which are subsequently transformed to between-taxa distance matrices. These pairwise matrices are used to construct neighbour-joining trees for each of the top most-abundant clusters and they are finally summarized in a consensus network. This methodology was tested on three groups of angiosperms and one group of insects, resulting in congruent evolutionary hypotheses compared to more standard systematic analyses based on commonly used DNA markers. We propose that the combined application of these phylogenetic approaches based on repeat abundances and repeat sequence similarities could be helpful to understand mechanisms governing genome and repeatome evolution.


The effect of apricots on the experimental cataract model formed by sodium selenite.
This study was designed in order to investigate whether sun dried apricots have a preventive effect on the experimental cataract model formed by sodium selenite in rats. Fifty-nine Spraque-Dawley rat pups were divided into three groups. Group 1 (control group) consisted of twenty rat pups, born from the rats nourished ad libitum. Group 2 consisted of 18 newborn rats, born from the rats nourished ad libitum with 10% sun dried natural apricots. Group 3 consisted of 21 newborn rats, born from the rats nourished ad libitum. Subcutaneous (30nmol/gr) sodium selenite injection was applied to all the newborn rats except the control group (Group 1) on postpartum day 10. Cataract development was graded by slit-lamp examination and photography. Encapsulated lenses were analyzed for reduced glutathione (GSH) and malondialdehyde (MDA), a marker of lipid per oxidation. Lenses were also analyzed for total nitrite (TN). The presence of oxidative stress in selenite cataract development and its prevention by sun dried apricots.


Design of pressure-driven microfluidic networks using electric circuit analogy.
This article reviews the application of electric circuit methods for the analysis of pressure-driven microfluidic networks with an emphasis on concentration- and flow-dependent systems. The application of circuit methods to microfluidics is based on the analogous behaviour of hydraulic and electric circuits with correlations of pressure to voltage, volumetric flow rate to current, and hydraulic to electric resistance. Circuit analysis enables rapid predictions of pressure-driven laminar flow in microchannels and is very useful for designing complex microfluidic networks in advance of fabrication. This article provides a comprehensive overview of the physics of pressure-driven laminar flow, the formal analogy between electric and hydraulic circuits, applications of circuit theory to microfluidic network-based devices, recent development and applications of concentration- and flow-dependent microfluidic networks, and promising future applications. The lab-on-a-chip (LOC) and microfluidics community will gain insightful ideas and practical design strategies for developing unique microfluidic network-based devices to address a broad range of biological, chemical, pharmaceutical, and other scientific and technical challenges.


Early progression of diabetic nephropathy correlates with methylglyoxal-derived advanced glycation end products.
Increased advanced glycation end products (AGEs) and oxidation products (OPs) are proposed to lead to progression of diabetic nephropathy (DN). We investigated the relationship between AGEs, OPs, and progression of DN in 103 subjects with type 1 diabetes participating in the Natural History of Diabetic Nephropathy Study. Mean age of subjects was 17.6±7.4 years, and mean duration of diabetes was 8.3±4.9 years. All patients were normoalbuminuric. Change in glomerular basement membrane (GBM) width from baseline to 5 years, measured using electron micrographs of renal biopsies, was our primary end point, and mesangial fractional volume was a secondary end point. Fast progressors (FPs) were defined as those in the upper quartile of GBM change, and the remaining patients were classified as slow progressors (SPs). AGEs (3-deoxyglucosone and methylglyoxal hydroimidazolones [MGHI]), carboxymethyl lysine (CML), carboxyethyl lysine (CEL), and OPs (methionine sulfoxide and 2-aminoadipic acid) were measured at year 5 by liquid chromatography/triple-quadruple mass spectroscopy on 10-K plasma filtrates. We found that MGHI, CEL, and CML levels were significantly higher in FPs relative to SPs. No product predicted mesangial expansion. A model containing only HbA1c accounted for 4.7% of GBM width variation, with the total variability explained by the model increasing to 11.6% when MGHI, CEL, and CML were added to the regression model (7.9% increase). MGHI was a significant independent predictor of FP. Using a logistic regression model to relate each biomarker to the probability of a subject's classification as an FP, CML, CEL, and MGHI, but not HbA1c, showed a significant relationship to the probability of FP. The results suggest that these three major AGEs may be early indicators of progression of important DN lesions.


A retrospective cohort study of economic outcomes and adherence to monotherapy with metformin, pioglitazone, or a sulfonylurea among patients with type 2 diabetes mellitus in the United States from 2003 to 2005.
The aims of this study were to compare all-cause total health care costs and diabetes mellitus (DM)-specific health care costs between patients who were adherent or nonadherent to monotherapy with metformin, pioglitazone, or a sulfonylurea and to examine whether cost differences varied among patients using these oral antidiabetic drugs. This was a retrospective cohort study using data from the MEDSTAT MarketScan Research Databases. Patients aged 18 to 90 years who were continuously insured between 2003 and 2005 and had > or =2 outpatient claims or > or =1 inpatient claim with a diagnosis of DM (International Classification of Diseases, Ninth Revision, Clinical Modification code 250.xx) in 2003 were eligible for the study. To be part of the final sample, patients had to fill > or =2 prescriptions for metformin, pioglitazone, or a sulfonylurea during 2003, including > or =1 prescription during the last 3 months of the year. Patients were not eligible if they were taking polytherapy or a combination drug. All eligible patients were followed in 2004 and 2005. Adherence was calculated for each year using a medication possession ratio, and was dichotomized at > or =80% as either adherent or nonadherent. Annual all-cause health care costs and diabetes-specific costs were estimated using generalized linear models, adjusting for demographic characteristics, insurance, and comorbid conditions. A total of 108,592 patients who met the inclusion criteria were identified. Their mean age was 63 years; 49.8% (54,037/108,592) were women. More pioglitazone users resided in the north-central or south regions (81.3% [9364/11,520]) compared with metformin (62.4% [32,550/52,156]) or sulfonylurea (62.6% [28,105/44,916]) users (P < 0.001). Mean comorbidity scores were higher in the sulfonylurea (1.78) and pioglitazone (1.69) group than in the metformin group (1.45) (P < 0.001). Mean adherence ranged from 61.3% to 73.8% during the 2 years of follow-up. After adjustment, all-cause health care costs were $12,412 annually among adherent patients and $13,258 among nonadherent patients (difference, $846 [95% CI, $747 to $945]). Diabetes-related health care costs were $2230 annually among adherent patients and $2284 among nonadherent patients (difference, $55 [95% CI, $33 to $77]). In specific monotherapy groups, adjusted annual all-cause health care costs were $336 higher (95% CI, $216 to $456) for nonadherent metformin users, $1140 higher (95% CI, $793 to $1486) for nonadherent pioglitazone users, and $1509 higher (95% CI, $1339 to $1679) for nonadherent sulfonylurea users compared with adherent users. Compared with metformin users, sulfonylurea and pioglitazone users had greater adherence-related differences in all-cause health care costs (P < 0.05). There was no significant difference in diabetes-specific total costs between nonadherent and adherent patients taking metformin (difference, $6; 95% CI, -$31 to $20). Patients who were nonadherent to sulfonylureas had $271 higher (95% CI, $235 to $307) diabetes-specific costs per year than patients who were adherent to sulfonylureas. Patients who were nonadherent to pioglitazone had $433 lower (95% CI, -$516 to -$350) diabetes-specific costs per year than patients who were adherent to pioglitazone. Adherence with metformin, pioglitazone, or a sulfonylurea was associated with overall cost reductions in the patients studied, but these cost reductions varied by monotherapy. Adherence to sulfonylureas or pioglitazone was associated with greater total cost reductions than was adherence to metformin. Health systems that commit resources to improving interventions may be able to achieve a return on investment if adherence to oral antidiabetic agents can be improved.


Successful Extubation After Weaning Failure by Noninvasive Ventilation in Patients With Neuromuscular Disease: Case Series.
To report successful cases of extubation from invasive mechanical ventilation at our institution using pulmonary rehabilitation consisting of noninvasive ventilation (NIV) in neuromuscular patients with experience of reintubation. Patients who experienced extubation failure via the conventional weaning strategy but afterwards had extubation success via NIV were studied retrospectively. Continuous end-tidal CO2 (ETCO2) and pulse oxyhemoglobin saturation (SpO2) monitoring were performed. Extubation success was defined as a state not requiring invasive mechanical ventilation via endotracheal tube or tracheotomy during a period of at least 5 days. A total of 18 patients with ventilatory failure who initially experienced extubation failure were finally placed under part-time NIV after extubation. No patient had any serious or long-term adverse effect from NIV, and all patients left the hospital alive. NIV may promote successful weaning in neuromuscular patients with experience of reintubation.


Comparative effectiveness of standard care with IV thrombolysis versus without IV thrombolysis for mild ischemic stroke.
One third of patients presenting with initially mild strokes have unfavorable outcomes, and the efficacy of intravenous thrombolysis (IVT) in this population has not been proven. This study aimed to evaluate the comparative effectiveness of standard care with IVT versus without IVT in mild stroke patients. Using a multicenter stroke registry database, we identified patients with acute ischemic stroke who presented within 4.5 hours of symptom onset and had initial National Institutes of Health Stroke Scale scores ≤5. Multivariable logistic analysis and propensity score matching were used to adjust for baseline imbalances between the patients who did and did not receive IVT. Adjusted odds ratios and 95% CIs of IVT were estimated for 3-month modified Rankin Scale scores of 0 to 1 and symptomatic. Of 13 117 patients with stroke who were hospitalized between April 2008 and May 2012, 1386 met the eligibility criteria, and 194 (14.0%) were treated with IVT. For a modified Rankin Scale of 0 to 1 at 3 months, the adjusted odds ratios were 1.96 (95% CI, 1.28 to 3.00; P=0.002) by multivariable logistic analysis and 1.68 (1.10 to 2.56; P=0.02) by propensity score matching analysis, respectively. There was a statistically nonsignificant excess of symptomatic hemorrhagic transformation (odds ratios=3.76 [0.95 to 16.42; P=0.06] and 4.81 [0.84 to 49.34; P=0.09]), respectively. In this observational registry-based study, standard care with IVT is more effective than not receiving IVT in mildischemic stroke patients, and there is a statistically nonsignificant risk of symptomatic hemorrhagic transformation.


Parathyroid score can predict the duration of required calcium supplementation after total thyroidectomy.
Postoperative hypoparathyroidism is the most common complication after total thyroidectomy, owing to unintentional injury or decreased blood flow to the parathyroid glands. Prediction of postoperative hypoparathyroidism would be helpful for surgeons to manage postoperative hypocalcemia. In this study, we scored the discoloration of the parathyroid glands using a new parathyroid scoring system and evaluated the correlation between the parathyroid score and duration of required calcium supplementation after total thyroidectomy. A total of 316 patients undergoing total thyroidectomy between November 2009 and April 2010 were enrolled in this retrospective study. Parathyroid scoring was performed by one experienced surgeon. The status of each of the 4 parathyroid glands was classified as normal color (3 points), slightly discolored (2 points), dark discoloration (1 point), or loss of the gland (0 points), resulting in possible total scores of 0-12. Serum parathyroid hormone (PTH), serum calcium, and ionized calcium concentrations were measured at 2 hours, 2 weeks, 3 months, 6 months, and 1 year after surgery. Patients were also divided into three groups based on the duration of required calcium supplementation: no required supplementation (n = 260, 82.3%), required supplementation for <6 months (n = 38, 12%), and required supplementation for ≥6 months (n = 18, 5.75%). Parathyroid scores were positively correlated with ionized PTH concentrations at 2 hours (r = 0.053, p < 0.001), 2 weeks (r = 0.056, p < 0.001), 3 months (r = 0.032, p<0.001), 6 months (r = 0.072, p < 0.001), and 1 year (r = 0.071, p < 0.001) after thyroidectomy. Parathyroid scores were significantly and inversely associated with the duration of required calcium supplementation (p = 0.001). Parathyroid scores at the end of surgery might be helpful for predicting the degree of postoperative hypocalcemia after total thyroidetomy.


[The gut-brain axis in attention deficit hyperactivity disorder: the role of the microbiota].
Attention deficit hyperactivity disorder (ADHD) has a complex aetiology, mainly attributed to a number of susceptibility genes and environmental factors. Genetic association studies, however, have been inconsistent and have identified genetic variants with a moderate effect that explain a small proportion of the estimated inheritability of the disorder (< 10%). Recent studies suggest that the gut microbiota and diet play an important role in the development and symptoms of different mental disorders. Nevertheless, no clear evidence exists on the issue. This project proposes an alternative approach to identify mechanisms by which the intestinal microbial ecosystem and diet could contribute to the presence of ADHD. To identify biomarkers for ADHD by examining the gut microbiota. We conducted a cross-sectional study of adult patients with ADHD (n = 100) and control subjects (n = 100). Measures of ADHD evaluation and eating habits were performed in both groups. Samples of faecal material were obtained from which to extract bacterial DNA, then used to characterise the participants' gut microbiota. A meta-genomic association study was later performed to attempt to correlate the bacterial composition of the intestine with the clinical subtypes of the disorder. Comparing the gut microbiota profiles of subjects with ADHD and controls is expected to help account for the clinical heterogeneity of the disorder and identify new mechanisms involved in its development.


At any level of adiposity, relatively elevated leptin concentrations are associated with decreased insulin sensitivity.
The impact of obesity on glucose homeostasis has a high interindividual variability, which may be partially explained by different adipokines concentrations. Leptin regulates energy balance and metabolism, and although its plasma levels are proportional to fat mass, they vary significantly across individuals with the same level of adiposity. We tested whether glucose homeostasis differs in subjects with similar degrees of adiposity but different leptin levels. We analyzed 1,290 healthy adults from the RISC study cohort (30-60 years, M/F 577/713, BMI 25 ± 3 kg/m2) characterized for body composition and metabolic variables with a 75-g OGTT, euglycemic-hyperinsulinemic clamp, β-cell function and lipidomics. Individuals were divided into relatively high and low leptin (RHL and RLL) if they were above or below the sex-specific leptin-fat mass(%) regression. Despite similar glucose tolerance, RHL showed markedly higher fasting and OGTT insulin concentration (+30% and +29% respectively, p < 0.0001) and secretion (+17% and +11% respectively, p < 0.0001). Regardless of BMI, RHL individuals had lower whole-body (-17-23%, p < 0.0001) and adipose tissue insulin sensitivity (-24%, p < 0.0001) compared to RLL. Notably, lean RHL individuals showed similar insulin sensitivity and β-cell function to RLL individuals with overweight/obesity. Subjects with leptin levels that are inappropriately elevated for their fat mass show whole-body/adipose tissue insulin resistance and hyperinsulinemia, regardless of BMI.


[Laser photocoagulation in treatment of plexiform neurofibromas in maxilla-facial area].
The incidence of plexiform neurofibromas (PN) is 1:2500-3000, the disease affects maxilla-facial area in 31% of cases. The current paper describes results of treatment of 23 patients as well as preliminary data on using laser coagulation as treatment option for PN, namely on preoperative stage of treatment.


Disequilibrium in the triose phosphate isomerase system in rat liver.
1. The equilibrium constant at 38 degrees and I 0.25 of the triose phosphate isomerase reaction was found to be 22.0 and that of the aldolase reaction, 0.99x10(-4)m. The [dihydroxyacetone phosphate]/[glyceraldehyde phosphate] ratio was found to be 9.3 in rat liver. The causes of the apparent deviation of the triose phosphate isomerase system from equilibrium in vivo have been investigated. 2. The equilibria of the triose phosphate isomerase and aldolase reactions were studied with relatively large concentrations of crystalline enzymes and small concentrations of substrates, approximating to those found in rat liver and muscle. There was significant binding of fructose diphosphate by aldolase under these conditions. There was no evidence that binding of glyceraldehyde phosphate by either enzyme affected the equilibria. 3. The deviation from equilibrium of the triose phosphate isomerase system in rat liver can be accounted for by the low activity of the enzyme, in relation to the flux, at low physiological concentrations of glyceraldehyde phosphate (about 3mum). It has been calculated that a flux of 1.8mumoles/min./g. wet weight of liver would be expected to cause the measured degree of disequilibrium found in vivo. 4. The conclusion that the triose phosphate isomerase is not at equilibrium is in accordance with the situation postulated by Rose, Kellermeyer, Stjernholm & Wood (1962) on the basis of isotope-distribution data. 5. The triose phosphate isomerase system is closer to equilibrium in resting muscle probably because of a very low flux and a high enzyme concentration. 6. The aldolase system deviated from equilibrium in rat liver by a factor of about 10 and by a much greater factor in resting muscle. 7. The measurement of total dihydroxyacetone phosphate and glyceraldehyde phosphate content indicates the concentrations of the free metabolites in the tissue. This may not hold for fructose diphosphate, a significant proportion of which may be bound to aldolase.


Effect of supporter characteristics on expression of negative social reactions toward rape survivors in Dar Es Salaam, Tanzania.
Using a community representative sample of 1,505 adults we examined interpretations of rape situations in order to establish attitudes toward rape. We assessed their intentions to express negative social reactions (NSRs) toward rape survivors. We then determined effects of attitudinal and sociodemographic characteristics in logistic regression models assessing the odds of expressing NSRs. Being old, male, and Muslim, and failing to interpret the legal circumstances of rape increased their risks of expressing NSRs. The degree of interpretation of lack of consent as rape affected their intentions to express NSRs, but not how they responded to survivors of different social status.


The adaptor protein Sh2d3c is critical for marginal zone B cell development and function.
Sh2d3c is an adaptor protein that has been implicated in T cell activation and shown to associate with different components of the integrin signaling pathway ex vivo. However, the in vivo significance of Sh2d3c expression in the regulation of the immune response and/or hematopoietic cell lineage development is not known. In this study, we show that expression of Sh2d3c is more critical for development and function of marginal zone B (MZB) cells than for T cell maturation. Mice deficient in Sh2d3c expression (Sh2d3c(-/-)) had a reduced number of MZB cells, and the residual MZB cells failed to properly capture polysaccharide Ags. Activation-induced proliferation, cytokine production, and migration of Sh2d3c(-)(/)(-) splenic B cells were also significantly reduced in vitro compared with wild-type (Sh2d3c(+/+)) cells. In contrast, T cell development and function were largely normal in Sh2d3c(-/-) mice. The thymi of Sh2d3c(-/-) mice showed no maturational abnormalities, the number of splenic T cells was only modestly reduced, and the T cells responded normally to in vitro polyclonal activation. The observed B cell deficiency in the Sh2d3c(-/-) mice led to diminished humoral immune response against thymus-independent type 2, but not thymus-dependent Ags, which highlights the primary in vivo role of Sh2d3c in regulating B cell development and function.


Serum ferritin as a tumor marker in patients with squamous cell carcinoma of the head and neck.
A double antibody enzyme immunoassay was used to measure serum ferritin levels in several different control and tumor-bearing populations collected from two institutions. The control groups consisted of normal volunteers, smokers, and Latter Day Saints. No statistically significant differences were noted in ferritin levels between pairs of these groups. Differences were noted among the normal groups when separated on the basis of age and sex, with higher ferritin levels in individuals older than 32 years of age and in men. By one-way analysis of variance, most control groups and subgroups were shown to have significantly lower levels (P less than 0.05) than the head and neck cancer group, with the exception of male smokers, who had levels comparable to male head and neck cancer patients. Serum ferritin levels were higher in head and neck cancer patients than in controls; however, there was no difference when compared with patients with other types of solid malignancies or when considering the anatomic site of the head and neck lesion. Ferritin levels were significantly (P less than 0.05) higher in patients with advanced (Stages III and IV) cancer than in those individuals with Stage I or II cancer. In patients with no evidence of clinical disease 5 years after treatment, the ferritin level had essentially returned to normal. In a group of head and neck cancer patients followed longitudinally, a significant decline in ferritin levels (P less than 0.05) was seen by 5 months after the completion of successful treatment. Furthermore, ferritin levels showed a tendency to increase or remain at high levels in patients with a poor prognosis and to decrease in those patients with a favorable prognosis, giving support to the contention that ferritin may prove to be a valuable tumor marker in head and neck cancer.


Epithelial cell-turnover ensures robust coordination of tissue growth in Drosophila ribosomal protein mutants.
Highly reproducible tissue development is achieved by robust, time-dependent coordination of cell proliferation and cell death. To study the mechanisms underlying robust tissue growth, we analyzed the developmental process of wing imaginal discs in Drosophila Minute mutants, a series of heterozygous mutants for a ribosomal protein gene. Minute animals show significant developmental delay during the larval period but develop into essentially normal flies, suggesting there exists a mechanism ensuring robust tissue growth during abnormally prolonged developmental time. Surprisingly, we found that both cell death and compensatory cell proliferation were dramatically increased in developing wing pouches of Minute animals. Blocking the cell-turnover by inhibiting cell death resulted in morphological defects, indicating the essential role of cell-turnover in Minute wing morphogenesis. Our analyses showed that Minute wing discs elevate Wg expression and JNK-mediated Dilp8 expression that causes developmental delay, both of which are necessary for the induction of cell-turnover. Furthermore, forced increase in Wg expression together with developmental delay caused by ecdysone depletion induced cell-turnover in the wing pouches of non-Minute animals. Our findings suggest a novel paradigm for robust coordination of tissue growth by cell-turnover, which is induced when developmental time axis is distorted.


Conformity in diversity? Isotopic investigations of infant feeding practices in two iron age populations from Southern Öland, Sweden.
This article presents the results of a study of infant diet at two Iron Age sites on the island of Öland, Sweden. The cemetery at Bjärby contained a large number of subadults who had survived the earliest years of life, whereas most individuals at Triberga had died by 6 months of age. To investigate whether differences in infant feeding could explain the different mortality rates, the carbon, nitrogen, and sulfur stable isotope ratios of bone and tooth dentin collagen from the two sites were analyzed. Twenty-two samples from Triberga and 102 from Bjärby yielded data that could be included in the carbon and nitrogen analysis. Twelve samples from Triberga and 42 from Bjärby were included in the sulfur analysis. The results for carbon (δ(13) C: Triberga X = -18.8, s.d. = 1.1; Bjärby X = -19.8, s.d. = 0.4), nitrogen (δ(15) N: Triberga X = 12.9, s.d. = 1.5; Bjärby X = 13.4, s.d. = 1.4), and sulfur (δ(34) S: Triberga X = 8.1, s.d. = 1.1; Bjärby X = 5.8, s.d. = 1.3) suggest that diet was broadly similar at both sites and based on terrestrial resources. At Bjärby, females and high-status individuals consumed higher-trophic level protein than other males from early childhood onward. There was some indication that the contribution of marine resources to the diet may also have differed between the sexes at Triberga. No consistent differences in breast milk intake were observed between the two sites, but there was substantial variation at each. This variation may reflect an influence of gender and social status on infant feeding decisions.


A randomized trial of mifepristone (10 mg) and levonorgestrel for emergency contraception.
To compare the efficacy, patient acceptability and adverse effects of low-dose mifepristone (10 mg) with the levonorgestrel regimen (2 doses of 750 microg given 12 hours apart) for emergency contraception. This randomized controlled trial compared mifepristone (10 mg) to levonorgestrel (2 doses of 750 microg given 12 hours apart) in the context of emergency contraception within 120 hours of unprotected intercourse. The primary outcome measure was unintended pregnancy. Secondary outcomes included adverse effects experienced by women, acceptability of the method of emergency contraception used, and the timing of the first menstrual cycle after treatment. The total number of women recruited was 2,065. The crude pregnancy rates were 1.3% and 2.0% for mifepristone and levonorgestrel (P = .46), with 77% and 64% of expected pregnancies prevented, respectively. Women receiving mifepristone were more likely to have a delayed onset of the subsequent menstrual cycle after treatment (P < .001), whereas those having levonorgestrel were more likely to have an early onset of the subsequent menstrual cycle (P < .001). Acceptability levels were high for both methods, with 94% of women receiving mifepristone and 91% receiving levonorgestrel expressing satisfaction. There was no difference in adverse effects (nausea, vomiting, breast tenderness, abdominal pain, lethargy, headache, hot flushes, and dizziness) experienced by women in the 2 groups. This study suggests that a small dose of mifepristone is not less effective than levonorgestrel for emergency contraception. Both regimens were highly acceptable to women.


Different recovery methods and muscle performance after exhausting exercise: comparison of the effects of electrical muscle stimulation and massage.
In this study we assessed the influence of the three different recovery interventions massage (MSG), electrical muscle stimulation (EMS), and passive rest (PR) on lactate disappearance and muscle recovery after exhausting exercise bouts. Twelve healthy male sport students participated in the study. They attended the laboratory on five test days. After measurement of [Formula: see text]O2max and a baseline Wingate test (WGb), the three recovery interventions were tested in random counterbalanced order. High intensity exercise, which consisted of six exhausting exercise bouts (interspersed with active recovery), was followed by MSG, EMS or PR application (24 minutes); then the final Wingate test (WGf) was performed. Lactate, heart rate, peak and mean power, rating of perceived exertion (RPE), and total quality of recovery (TQR) were recorded. In WGf mean power was significantly higher than in WGb for all three recovery modalities (MSG 6.29%, EMS 5.33%, PR 4.84% increase, p < 0.05), but no significant differences in mean and peak power were observed between the three recovery modes (p > 0.05). The heart rate response and the changes in blood lactate concentration were identical in all three interventions during the entire protocol (p = 0.817, p = 0.493, respectively). RPE and TQR scores were also not different among the three interventions (p > 0.05). These results provide further evidence that MSG and EMS are not more effective than PR in the process of recovery from high intensity exercise.


From farm to fork: Spread of a multidrug resistant Salmonella Infantis clone encoding blaCTX-M-1 on pESI-like plasmids in Central Italy.
Salmonella enterica subsp. enterica serovar Infantis (S. Infantis) is one of the "top five Salmonella serovars" of clinical significance in the European Union (EU). Antimicrobial resistant and extended spectrum β-lactamase (ESBL) AmpC-producing S. Infantis have been described in food production systems and human clinical samples in Italy. Recently, an increase of MDR S. Infantis carrying blaCTX-M genes involved in 3rd generation cephalosporin resistance was noticed in the EU, including Italy, mainly due to the spread of S. Infantis harboring a pESI-like plasmid. The aim of this study was to investigate the occurrence of the S. Infantis pESI-like plasmid among antibiotic resistant S. Infantis strains isolated at different points of the food chain, and to provide a phylogenetic analysis to gain further insight on their transmission pathways from 'farm to fork'. MDR S. Infantis strains (n. 35) isolated from 2016 to 2021 at different stages of the food chain (animals, food, food-related environments, and humans) were investigated with in depth molecular characterization using real-time PCR, S1 nuclease pulsed-field gel electrophoresis (S1-PFGE) and whole genome sequencing (WGS). Our study reported the occurrence of S. Infantis strains harboring pESI-like plasmids, carrying blaCTX-M-1 genes, in Central Italy, at different sampling points along the food chain. Results confirmed the presence of a plasmid with a molecular size around 224-310 kb, thus consistent with the pESI-like, in 97 % of the 35 samples investigated. Two variants of S. Infantis pESI-like IncFIB(K)_1_Kpn3 were detected, one associated with the European clone carrying blaCTX-M-1 (21 isolates) and the other associated with U.S. isolates carrying blaCTX-M-65 (2 isolates, pESI-like U.S. variant). The majority was resistant to 3rd generation cephalosporins but none of the strains tested positive for the carbapenemase encoding genes. A total of 118 virulence genes were identified in isolates harboring the pESI-like plasmid. cgMLST and SNP-based analysis revealed the presence of one main cluster, composed by strains isolated from the environment, animals, food and humans. The results of this investigation underline the importance of phylogenetic studies to monitor and understand pathogen and AMR spread in a One Health approach.


Soluble sugar-based quinoline derivatives as new antioxidant modulators of metal-induced amyloid aggregation.
Oxidative stress and protein aggregation have been demonstrated to be the major factors involved in neurodegenerative diseases. Metal ions play a pivotal role, acting as mediators of neurotoxicity either by favoring or redox cycling. Thus, they represent a promising and suitable therapeutic target for the treatment of neurodegenerative disorders. In particular, the development of bifunctional or multifunctional molecules, which have antiaggregant and metal-chelating/antioxidant properties, may be considered as a valuable strategy for the treatment of neurodegeneration considering its multifactorial nature. Herein, we report the design and the characterization of four new multifunctional sugar-appended 8-hydroxyquinolines focusing on the effects of the conjugation with trehalose, a nonreducing disaccharide involved in the protection of proteins and cells against environmental stresses. These glycoconjugates do not exhibit any antiproliferative activity against three human cell lines of different histological origin, unlike 8-hydroxyquinolines. The multiple properties of the new derivatives are highlighted, reporting their Cu(2+) and Zn(2+) binding ability, and antioxidant and antiaggregant capacities. In particular, these latter were determined by different assays, including the evaluation of their ability to modulate or even suppress the aggregation of Aβ1-40 and Aβ1-42 peptides induced by copper or zinc ions.


Laparoscopic subtotal colectomy with antiperistaltic cecorectal anastomosis: a new step in the treatment of slow-transit constipation.
Subtotal colectomy with antiperistaltic cecorectal anastomosis (SCCRA) has proved to be an effective alternative to total colectomy for the treatment of severe slow-transit constipation. The laparoscopic approach has made this procedure even more attractive. This is the first controlled trial on laparoscopic SCCRA. The study compares the laparoscopic and the open approach. Since 2001, all SCCRAs have been performed laparoscopically at our institution. Only severely symptomatic patients are offered surgery, after stringent patient selection. Laparoscopic SCCRA was performed following the same steps that we first described for the open approach, by utilizing a five-trocar technique. Outcome parameters were prospectively collected every 3 and 6 months. Wexner constipation and incontinence scales (WCS, WI) and gastrointestinal quality of life index (GIQLI) were adopted for functional results. We conducted a case-control study of 15 consecutive patients who underwent laparoscopic SCCRA (VL) and 15 patients previously operated on by the open approach (Op) to compare postoperative and functional outcomes. The VL group had better postoperative outcomes (pain, ileus) while complication rates were similar. Resolution of constipation was impressive in both groups, with no significant difference at follow-up. The VL group presented with a higher number of bowel movements at 3 months (3.8 vs. 2.8, p = 0.039), resulting in a significantly higher incontinence rate at 3 months (WI 6.4 vs. 2.73, p = 0.004), although the difference was no longer significant at 1-year follow-up. The quality of life was good for both groups; the VL group showed a significant improvement at 1-year follow-up (64.18 vs. 114.79, p < 0.01). Laparoscopic SCCRA confirmed the good functional results of the open approach, with no increase in morbidity rate and a faster postoperative recovery. An early higher incontinence rate did not affect quality of life.


Suicidal ideation and anxiety disorders: elevated risk or artifact of comorbid depression?
Research into the possible relationship between anxiety disorders and suicidal ideation has yielded mixed results, leading some to suggest that the positive findings between anxiety and suicidal ideation might simply be a by-product of comorbid depression. Recent work has suggested that having an anxiety disorder without history of mood disorder does convey increased risk for suicidal ideation, although the study could not assess for the possible impact of subsyndromal depressiveness. This current study, therefore, examined the relationship between anxiety disorder symptoms and suicidality using continuous scales and controlling for depressiveness. Data regarding the severity of panic, social anxiety, generalized anxiety, and obsessive-compulsive symptoms were obtained from a sample of 166 college students. Results generally supported the conclusions that anxiety disorders convey risk for suicidal ideation above and beyond any co-occurring depressiveness, and anxiety and depression together conveyed an additional interactive risk.


Testing Robustness of Child STEPs Effects with Children and Adolescents: A Randomized Controlled Effectiveness Trial.
A critical task in psychotherapy research is identifying the conditions within which treatment benefits can be replicated and outside of which those benefits are reduced. We tested the robustness of beneficial effects found in two previous trials of the modular Child STEPs treatment program for youth anxiety, depression, trauma, and conduct problems. We conducted a randomized trial, with two significant methodological changes from previous trials: (a) shifting from cluster- to person-level randomization, and (b) shifting from individual to more clinically feasible group-based consultation with STEPs therapists. Fifty community clinicians from multiple outpatient clinics were randomly assigned to receive training and consultation in STEPs (n= 25) or to provide usual care (UC; n= 25). There were 156 referred youths-ages 6-16 (M= 10.52, SD = 2.53); 48.1% male; 79.5% Caucasian, 12.8% multiracial, 4.5% Black, 1.9% Latino, 1.3% Other-who were randomized to STEPs (n= 77) or UC (n= 79). Following previous STEPs trials, outcome measures included parent- and youth-reported internalizing, externalizing, total, and idiographic top problems, with repeated measures collected weekly during treatment and longer term over 2 years. Participants in both groups showed statistically significant improvement on all measures, leading to clinically meaningful problem reductions. However, in contrast to previous trials, STEPs was not superior to UC on any measure. As with virtually all treatments, the benefits of STEPs may depend on the conditions-for example, of study design and implementation support-in which it is tested. Identifying those conditions may help guide appropriate use of STEPs, and other treatments, in the future.


Glyphosate skin binding, absorption, residual tissue distribution, and skin decontamination.
Glyphosate is a broad-spectrum postemergence translocated herbicide. Its interactions with skin and potential systemic availability through percutaneous absorption was studied by skin binding, skin absorption, residual tissue distribution, and skin decontamination. Glyphosate in a final formulation (Roundup) undiluted and diluted with water 1:20 and 1:32, would not partition into powdered human stratum corneum (less than 1%). In vitro percutaneous absorption through human skin into human plasma as receptor fluid was no more than 2% over a concentration range of 0.5-154 micrograms/cm2 and a topical volume range of 0.014-0.14 ml/cm2. Disposition of glyphosate following iv administration of 93 and 9 micrograms doses to rhesus monkeys was mainly through urine excretion, 95 +/- 8 and 99 +/- 4% in 7 days, respectively. Percutaneous absorption in vivo in rhesus monkey was 0.8 +/- 0.6% for the low dose (25 micrograms/cm2) and 2.2 +/- 0.8% for the high dose (270 micrograms/cm2). No residual 14C was found in organs of the monkeys euthanized 7 days after the topical application. Washing the skin application site with soap and water removed 90 +/- 4% of applied dose, and washing with water only removed 84 +/- 3% of applied dose. Both soap and water and water only were equal in ability to remove glyphosate from skin over a 24 hr skin application period. About 50% of the initially applied dose could be recovered after 24 hr. Glyphosate is very soluble in water and insoluble in most organics (octanol/water log P = -1.70) and therefore not compatible with the lipid-laden stratum corneum.(ABSTRACT TRUNCATED AT 250 WORDS)


Pharmacokinetics of trimethoprim-sulfamethoxazole in children.
The present report extends experience with the use of trimethoprim-sulfamethoxazole (TMP-SMX) in children aged 3 months to 10 years. The regimen was TMP (200 mg)--SMX (1000 mg)/m-2d given in two equal doses. The drug was easily administered, well tolerated and efficacious in the treatment of a variety of infections in 12 children. A steady state had been achieved by the third dose of medication and accumulation of either component during days 1 through 4 did not occur. Serum concentrations of TMP were slightly lower in children aged less than 3 years compared with those aged 3 to 6 years but the differences were small and these results are preliminary. Peak mean serum TMP concentration was highest at day 3 when it reached 1.63 mug/ml. It is concluded that this regimen may be suboptimal for some major parenchymal infections even though the therapeutic result was excellent in most children.


A simple method for assessing quinine pre-treatment in acute malaria.
Administration of a loading dose of quinine in severe malaria may be dangerous if therapeutic blood concentrations are already present because of previous treatment. To assess the reliability of the history of pretreatment we conducted a prospective study of 379 adult patients with acute falciparum malaria admitted to a hospital in western Thailand. Admission plasma concentrations of quinine were measured by high performance liquid chromatography (HPLC), and compared with the patients' history of previous quinine treatment. The sensitivity of the history was 59% (95%) confidence interval [CI] 49-69%), the specificity was 79% (95% CI 74-84%), the positive predictive value was 53%, and the negative predictive value 82%. A rapid (10 min semi-quantitative estimate of plasma quinine concentrations, based on simple 'dipstick' method using a quinine-specific monoclonal antibody, proved considerably more sensitive and specific. The correlation between the dipstick estimate and the subsequent HPLC measurement of plasma quinine concentration was 0.85 (n = 404; P < 0.0001). In 404 admission samples, the negative predictive value of the dipstick estimate for plasma quinine concentrations > 1 microgram/mL was 100%. In Thailand the history of previous quinine treatment given by the patients or their relatives was unreliable but the quinine dipstick provided a simple and rapid means of assessment of quinine pre-treatment in acute falciparum malaria.


Serum hyaluronidase levels in patients with aneurysmal subarachnoid haemorrhage.
The purpose of this study was to investigate the time course(s) of the serum hyaluronidase levels in patients with aneurysmal subarachnoid haemorrhage and to show whether there is a correlation between symptomatic vasospasm and serum levels of hyaluronidase. This prospective, open, non-randomised clinical study consisted of 20 patients with aneurysmal subarachnoid haemorrhage, and eight patients with normotensive hydrocephalus who served as the control group. Serum hyaluronidase levels were detected within the first three days, days five and seven after aneurysmal subarachnoid haemorrhage, and the results were compared with those from the control group. The results were also compared with those of the clinical parameters, including the patient's outcome at six months and symptomatic vasospasm. Mean serum hyaluronidase levels were higher on days five and seven, and comparisons with either day five (p-value is 0.001) and/or day seven (p-value is 0.00001) showed a statistical difference between subarachnoid haemorrhage and controls. However, no relationship was found between elevated serum hyaluronidase levels and the clinical parameters including symptomatic vasospasm (p-value is greater than 0.05) and outcome at sixth months (p-value is greater than 0.05). Our results indicate that serum hyaluronidase is elevated in the acute stage(s) of subarachnoid haemorrhage; however, no difference was found between serum hyaluronidase levels and subarachnoid haemorrhage severity. Clinical studies with larger population of patients with aneurysmal subarachnoid haemorrhage are required.


Effect of chondroitin sulfate-E on the osteoclastic differentiation of RAW264 cells.
The present study was designed to investigate whether chondroitin sulfate (CS)-E, a CS structural isomer variant, alter the differentiation of macrophage cell line RAW264 cells to osteoclast-like cells. CS-B, CS-E, low molecular weight CS-E, synthetic chondroitin polysulfate (CPS) and heparin significantly inhibited the formation of tartrate-resistant acid phosphatase-positive multinuclear cells and pit formation on calcium phosphate (CaP)-coated plates. CS-E pre-coated on the CaP plate also inhibited pit formation. Digestion of CS on the cell surface by chondroitinase showed no effect on the osteoclastic differentiation of RAW264 cells whereas inhibitory effect on the differentiation of osteoblastic cell line MC3T3-E1. On the other hand, exogenously added fluorescein-labeled CS-E directory bound to fibronectin and RAW264 cells. These results suggest that CS-E structure on the surface of osteoblasts or bone matrix binds to cell adhesion molecule such as integrin on the pre-osteoclastic cells and inhibits the differentiation into osteoclasts. CS-E may have a potential in treating bone defect if combined with CaP materials.


Impact of age on anaesthesiologists' competence: A narrative review.
The international anaesthesia community is getting older, in line with trends worldwide, and as men and women age there is the risk that psychophysiological decline could have an impact on clinical practice. Impairment of technical and nontechnical skills could have a negative impact on patients' safety and outcomes. The ageing process may not necessarily go hand-in-hand with a predictable pattern of decreased competence as not all aspects of functional decline are affected at the same rate and to the same extent. The development of simulation has provided a means of detecting and perhaps reversing the decline in ability associated with age. The introduction of recertification based on an assessment of competence at simulation sessions could play a crucial role in maintaining a high standard of patient care and an appropriate level of patient safety.


Ubiquitination of the Dishevelled DIX domain blocks its head-to-tail polymerization.
Dishevelled relays Wnt signals from the plasma membrane to different cytoplasmic effectors. Its signalling activity depends on its DIX domain, which undergoes head-to-tail polymerization to assemble signalosomes. The DIX domain is ubiquitinated in vivo at multiple lysines, which can be antagonized by various deubiquitinases (DUBs) including the CYLD tumour suppressor that attenuates Wnt signalling. Here, we generate milligram quantities of pure human Dvl2 DIX domain mono-ubiquitinated at two lysines (K54 and K58) by genetically encoded orthogonal protection with activated ligation (GOPAL), to investigate their effect on DIX polymerization. We show that the ubiquitination of DIX at K54 blocks its polymerization in solution, whereas DIX58-Ub remains oligomerization-competent. DUB profiling identified 28 DUBs that cleave DIX-ubiquitin conjugates, half of which prefer, or are specific for, DIX54-Ub, including Cezanne and CYLD. These DUBs thus have the potential to promote Dvl polymerization and signalosome formation, rather than antagonize it as previously thought for CYLD.


Common mitochondrial DNA deletion associated with sudden natural death in adults.
One of the most frequent causes of death in developed countries is sudden natural death (SND), which is the most common indication for medico-legal autopsies. Cardiac diseases are frequently detected among SND. Mitochondrial DNA (mtDNA) is easily damaged by reactive oxygen species, and it may cause dysfunction in tissues, leading to early events in cardiovascular disease. A specific mtDNA deletion of 4977 bp is associated to aging, myocardial dysfunction, and bioenergetic deficit. The potential link between mtDNA damage and SND has not been investigated before. Our aim was to evaluate the accumulation of the common mtDNA4977-deletion in cardiac muscle samples from autopsies of SND in adults (n = 14) in comparison to control samples from unnatural deaths (n = 12). Serial dilution-polymerase chain reaction method was performed to estimate the proportion of the total mtDNA harboring the mtDNA4977-deletion. Coefficient variation intra-assay was 8%, and inter-assay was 12%. MtDNA4977-deletion percentage was higher in samples obtained from victims of SND than in those from subjects who died of unnatural causes (p < 0.05). No differences in mtDNA4977-deletion were found between SND victims 39-51 years old, and no correlation was found between these samples and age, r = 0.30, p = 0.29 while it was significant among control samples, r = 0.68, p < 0.05. The association between mtDNA4977 deletion with SND victims might offer a tool to provide additional information to clarify complex SND investigations.


Studies on a central site of action for naloxone in endotoxin shock.
A complete understanding of naloxone's role in shock has not been well established; however, it has been accepted that naloxone applied specifically to the central nervous system can improve the course of the cardiovascular response. The currently available data would suggest that these central effects are mediated via nuclei that 1) are found in both the rostral and caudal regions of the brainstem, 2) contain opiate receptors, 3) interact with the fastigial nuclei of the cerebellum, and 4) possibly receive input from peripheral thermoreceptors. Recordings of neural activity from specific nuclei during shock as well as microinjections and iontophoresis of naloxone onto specific sites is needed to improve the current understanding of this apparently complex process.


Analysis of Chromatin Accessibility Changes Induced by BMMC Recognition of Foot-and-Mouth Disease Virus-like Particles through ATAC-seq.
Mast cells can recognize foot-and-mouth disease virus-like particles (FMDV-VLPs) via mannose receptors (MRs) to produce differentially expressed cytokines. The regulatory role of chromatin accessibility in this process is unclear. Bone marrow-derived mast cells (BMMCs) were cultured, and an assay of transposase-accessible chromatin sequencing (ATAC-seq) was applied to demonstrate the regulation of chromatin accessibility in response to the BMMCs' recognition of FMDV-VLPs. A pathway enrichment analysis showed that peaks associated with the nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), phosphatidylinositol 3 kinase-protein kinase B (PI3K-Akt), and other signaling pathways, especially the NF-κB pathway, were involved in the BMMCs' recognition of VLPs. Moreover, transcription factors including SP1, NRF1, AP1, GATA3, microphthalmia-associated transcription factor (MITF), and NF-κB-p65 may bind to the motifs with altered chromatin accessibility to regulate gene transcription. Furthermore, the expression of NF-κB, interleukin (IL)-9, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in the BMMCs of the VLP group increased compared with that of the BMMCs in the control group, whereas the expression of IL-10 did not differ significantly between groups. After inhibiting the MRs, the expression of NF-κB, IL-9, TNF-α, and IFN-γ decreased significantly, whereas the expression of IL-10 increased. The expression of MAPK and IL-6 showed no significant change after MR inhibition. This study demonstrated that MRs expressed on BMMCs can affect the NF-κB pathway by changing chromatin accessibility to regulate the transcription of specific cytokines, ultimately leading to the differential expression of cytokines. These data provide a theoretical basis and new ideas for the development of a novel vaccine for FMD.


Dynamic Contrast-Enhanced Ultrasound Radiomics for Hepatocellular Carcinoma Recurrence Prediction After Thermal Ablation.
To develop a radiomics model based on dynamic contrast-enhanced ultrasound (CEUS) to predict early and late recurrence in patients with a single HCC lesion ≤ 5 cm in diameter after thermal ablation. We enrolled patients who underwent thermal ablation for HCC in our hospital from April 2004 to April 2017. Radiomics based on two branch convolution recurrent network was utilized to analyze preoperative dynamic CEUS image of HCC lesions to establish CEUS model, in comparison to the conventional ultrasound (US), clinical, and combined models. Clinical follow-up of HCC recurrence after ablation were taken as reference standard to evaluate the predicted performance of CEUS model and other models. We finally analyzed 318 patients (training cohort: test cohort = 255:63). The combined model showed better performance for early recurrence than CUES (in training cohort, AUC, 0.89 vs. 0.84, P < 0.001; in test cohort, AUC, 0.84 vs. 0.83, P = 0.272), US (P < 0.001), or clinical model (P < 0.001). For late recurrence prediction, the combined model showed the best performance than the CEUS (C-index, in training cohort, 0.77 vs. 0.76, P = 0.009; in test cohort, 0.77 vs. 0.68, P < 0.001), US (P < 0.001), or clinical model (P < 0.001). The CEUS model based on dynamic CEUS radiomics performed well in predicting early HCC recurrence after ablation. The combined model combining CEUS, US radiomics, and clinical factors could stratify the high risk of late recurrence.


How many radiographs are needed to detect angular stable head screw cut outs of the proximal humerus - a cadaver study.
Cut out of locking head screws is the most common complication of locking plates in fracture fixation of the proximal humerus with potentially disastrous consequences. Aim of the study was to find the single best and combination of radiographic projections to reliably detect screw cut outs. The locking plate was fixed to six cadaveric proximal humeri. Six different radiographs were performed: anteriorposterior in internal (apIR), in neutral (ap0) and in 30° external rotation (apER); axial in 30° (ax30) and 60° (ax60) abduction and an outlet view. Each head screw (n=9) was sequentially exchanged to perforate the humeral head with the tip and all radiographs were repeated for each cut out. Randomized image reading by two blinded examiners for cut out was done for single projection and combinations. Interrater agreement was 0.72-0.93. Best single projection was ax30 (sensitivity 76%) and the worst was the outlet view (sens. 17%). Standard combination of apIR/outlet reached a sens. of 54%. The best combination of two was: apER/ax30 (90% sens.), of three: apIR/apER/ax30 (96% sens.) and of four: apIR/ap0/apER/ax30 (100% sens.). Standard radiographs (ap/outlet), especially in internal rotation, may miss nearly half of screw cut outs. Single best radiographic projection was an axial view with 30° abduction. To account for all cut outs and correct screw position a combination of four projections was needed. These simple and feasible intraoperative and postoperative radiographs help to detect screw perforations of the locking plate reliably. I (Study of Diagnostic Test).


Pathogenicity of a Human Laminin β2 Mutation Revealed in Models of Alport Syndrome.
Pierson syndrome is a congenital nephrotic syndrome with eye and neurologic defects caused by mutations in laminin β2 (LAMB2), a major component of the glomerular basement membrane (GBM). Pathogenic missense mutations in human LAMB2 cluster in or near the laminin amino-terminal (LN) domain, a domain required for extracellular polymerization of laminin trimers and basement membrane scaffolding. Here, we investigated an LN domain missense mutation, LAMB2-S80R, which was discovered in a patient with Pierson syndrome and unusually late onset of proteinuria. Biochemical data indicated that this mutation impairs laminin polymerization, which we hypothesized to be the cause of the patient's nephrotic syndrome. Testing this hypothesis in genetically altered mice showed that the corresponding amino acid change (LAMB2-S83R) alone is not pathogenic. However, expression of LAMB2-S83R significantly increased the rate of progression to kidney failure in a Col4a3-/- mouse model of autosomal recessive Alport syndrome and increased proteinuria in Col4a5+/- females that exhibit a mild form of X-linked Alport syndrome due to mosaic deposition of collagen α3α4α5(IV) in the GBM. Collectively, these data show the pathogenicity of LAMB2-S80R and provide the first evidence of genetic modification of Alport phenotypes by variation in another GBM component. This finding could help explain the wide range of Alport syndrome onset and severity observed in patients with Alport syndrome, even for family members who share the same COL4 mutation. Our results also show the complexities of using model organisms to investigate genetic variants suspected of being pathogenic in humans.


The levels of nitrite, nitrate and lipid peroxidation in diabetic mouse brain: the effect of melatonin and pentoxifylline.
This study investigated the relationship between diabetes (DM) and nitrite, nitrate and MDA levels and effect of melatonin and pentoxifylline. Sixty mice were randomly divided into four groups. Control: no action; Diabetes group (DM): after fasting-blood-glucose (FBG) was measured, 150 mg/kg alloxane was applied intraperitoneally three-times every other day; Diabetes + Melatonin (DM + MLT) and Diabetes + Pentoxifylline groups (DM + PTX): following the same procedures with DM, 10 mg/kg melatonin and 50 mg/kg pentoxifylline were administered subcutaneously six days, respectively. Following FBG analysis, brain tissues were taken under the anaesthesia. Nitrite, nitrate and MDA levels were measured. In the all groups with alloxane, FBG were higher than in before application (p < .05). Also, FBG, nitrite, nitrate and MDA levels in the DM + MLT and DM + PTX groups were lower than in the DM (p < .05). Nitrite and nitrate may be related to etiopathogenesis of DM, and pentoxifylline and especially melatonin relatively decrease nitrite, nitrate and lipid peroxidation.


Validation of El-Minia equation for estimation of glomerular filtration rate in different stages of chronic kidney disease.
Assessment of glomerular filtration rate (GFR) is the best index of kidney functions. We aimed to determine the accuracy of El-Minia equation for estimation of GFR in different stages of chronic kidney disease (CKD) in the El-Minia population of CKD patients, in comparison to other proposed equations. The study included 320 isotopic GFR measurements by 99m-Tc technetium diethylene triamine pentaacetic acid. Patients were categorized into the five CKD stages based on this measurement, and estimated GFRs based on the El-Minia, Cockroft-Gault, Chronic Kidney Disease Epidemiology Collaboration, Modification of Diet in Renal Disease (MDRD), abbreviated MDRD, Walser, Nankivell, Cockcroft-Gault, and Mayo Clinic equations were validated using the National Kidney Foundation guidelines. The El-Minia Equation showed the best performance in all 5 stages of CKD, except for stage 1, in which the Cockroft-Gault equation showed the best performance (44% within ± 10% error). In addition, the El-Minia equation provided the best accuracy to classify CKD stages, except for stage 4, in which the MDRD gave the best accuracy (82%). Worsening of the accuracy was documented in all the eight equations with more severe stages of kidney disease. These results were not significantly changed when the analyses were restricted to diabetic patients. The El-Minia equation represents a better estimation of GFR in all stages of CKD than other published equations for CKD patients in El-Minia, Egypt. This equation was the best one for classification of CKD in these patients.


[The efficacy of omalizumab in the treatment of chronic rhinosinusitis with nasal polyps: a discussion of 2 refractory cases].
Nasal polyposis is a specific phenotype of chronic rhinosinusitis (CRS). Some cases can be managed with topical and infrequent use of systemic steroids, while many patients require surgery. Despite postoperative, regular steroid administration, recurrences may be found especially in patients suffering from Aspirin exacerbated respiratory disease (AERD), a particularly severe form of CRS with polyps, asthma and non-steroid-anti-inflammatory-drug (NSAID) intolerance. We report two cases of difficult-to-treat AERD patients following revision surgery, treated with monoclonal anti-IgE antibody (omalizumab) and successful control of the disease and symptoms. Omalizumab may be a promising alternative in selected cases of CRS with nasal polyps to avoid overuse of systemic steroids and frustrating repetition of paranasal sinus surgeries.


Two case reports of mixed epithelial and stromal renal tumor of kidney in children and literature review analysis.
Mixed epithelial and stromal tumor of the kidney (MESTK) occurs almost exclusively in perimenopausal women while rarely in children. Only five pediatric patients have been described previously. Herein, we report a girl and a boy with MESTK, aged 3- and 4-years-old, respectively. The two patients presented with hematuria or an abdominal mass. Histologically, the tumors were both composed of epithelial and stromal elements. Immunohistochemical staining of tumor cells expressed epithelial and mesenchymal component markers. They were diagnosed with MESTK by histology and immunohistochemistry after surgery. The patients were at good condition after surgery. To our knowledge, these are the youngest reported cases of MESTK. And the glandular luminal structure formed by the epithelium was lined with urothelium, which expanded the histological map of MESTK.


Wnt3a signal pathways activate MyoD expression by targeting cis-elements inside and outside its distal enhancer.
Wnt proteins are secreted cytokines and several Wnts are expressed in the developing somites and surrounding tissues. Without proper Wnt stimulation, the organization of the dermomyotome and myotome can become defective. These Wnt signals received by somitic cells can lead to activation of Pax3/Pax7 and myogenic regulatory factors (MRFs), especially Myf5 and MyoD. However, it is currently unknown whether Wnts activate Myf5 and MyoD through direct targeting of their cis-regulatory elements or via indirect pathways. To clarify this issue, in the present study, we tested the regulation of MyoD cis-regulatory elements by Wnt3a secreted from human embryonic kidney (HEK)-293T cells. We found that Wnt3a activated the MyoD proximal 6.0k promoter (P6P) only marginally, but highly enhanced the activity of the composite P6P plus distal enhancer (DE) reporter through canonical and non-canonical pathways. Further screening of the intervening fragments between the DE and the P6P identified a strong Wnt-response element (WRE) in the upstream -8 to -9k region (L fragment) that acted independently of the DE, but was dependent on the P6P. Deletion of a Pax3/Pax7-targeted site in the L fragment significantly reduced its response to Wnt3a, implying that Wnt3a activates the L fragment partially through Pax3/Pax7 action. Binding of β-catenin and Pax7 to their target sites in the DE and the L fragment respectively was also demonstrated by ChIP. These observations demonstrated the first time that Wnt3a can directly activate MyoD expression through targeting cis-elements in the DE and the L fragment.


A maintenance protocol for psoriasis plaques cleared by the 308 nm excimer laser.
Using the 308 nm excimer laser, to evaluate the effectiveness of tapering the dose frequency on maintaining the clearance of psoriasis plaques after at least 75% clearance of a target plaque was achieved. This prospective case series, conducted at a university-based dermatology practice, enrolled five adults with stable, mild-to-moderate plaque-type psoriasis vulgaris. Patients received 308 nm UVB doses to affected areas. Initial dosing was based on multiples of a pre-determined minimal erythema dose (MED). Delivered fluences were 100, 150, 200, 250, 300 and 350 mJ/cm(2), corresponding to MED levels of 1 through 6. Subsequent doses were based on response to treatment. Induction treatments were scheduled biweekly for a total of 15 treatments. After improvement of disease, tapering began as follows: one treatment per week for 4 weeks, one treatment every other week for 4 weeks, and one final treatment 4 weeks later for a total of seven treatments. The main outcome measure was maintenance of disease improvement as determined by the Psoriasis Area and Severity Index (PASI) score and percentage improvement of target lesion and total body disease during and after the laser therapy taper. A flare was defined as a 25% worsening of the percentage improvement. All five patients completed the protocol. All patients achieved > or = 75% clearance of target plaques. No flares were noted upon completing the first month of the taper in any patient, either per percentage improvement or per PASI scores. One month after the second month of the taper (total of six treatments), no flares were noted and four out of five patients had no flares per PASI scores. No patients, either at the end of induction or at any interval during the taper, met the definition of a 50% rebound of PASI scores. Adverse effects included erythema, blisters, hyperpigmentation and erosions. All were well tolerated. Our results reaffirm that biweekly UVB laser treatments promote clearance of psoriatic plaques. A tapering of laser treatments may be beneficial in maintaining the level of plaque clearance obtained from biweekly laser treatments. Randomized, prospective trials are needed to establish laser therapy protocols for maintaining the plaque clearance achieved with the 308 nm excimer laser.


Enhanced recovery after bariatric surgery: Implementation of program and clinical outcomes from a bariatric surgery and metabolic diseases reference center.
Enhanced recovery after bariatric surgery (ERABS) protocols involve a series of multimodal perioperative procedures based on evidence designed to reduce physiological stress, improve recovery, and reduce costs on medical attention by decreasing length of hospital stay (length of stay [LOS]). The objective of the study was to report the viability and results of the ERABS application in a reference bariatric center. A prospective, observational, and descriptive study on bariatric procedures conducted over 12 months in the ERABS context which includes pre-procedure, intraprocedure, and post-procedure measures. The collected data include demographic data, comorbidity, morbimortality, LOS, and readmission to hospital. Sixty-four patients within a median of 38.8 ± 9.5 years and 44.1 ± 6.20 kg/m2 BMI underwent surgery. Fifty-five (85.93%) were Roux-en-Y gastric bypass (RYGB) cases and 9 (14.06%) were sleeve gastrectomy (SG). Related comorbidities were hypertension 37%, diabetes 34%, dyslipidemia 23%, and obstructive sleep apnea 21%. Two (3.12%) patients developed post-operative morbidity (respiratory and thromboembolic complications). LOS for RYGB was 1.16 ± 0.97 and 1 ± 0 days for SG. The successful discharge rate on the 1st day after procedure was 96% and 100%, respectively. Readmission to hospital within a 30-day period presented itself on 4 patients (6.25%). Applying ERABS protocols is feasible, safe, morbidity low, LOS acceptable, and a low readmission rate within 30 days.


[Morpho-functional characteristics of normal and pathological human megakaryocytes studied by selective silver staining of cell nucleoli].
The activity of nucleolar organizer (NO) in megakaryocytes (MG) from 8 donors, 10 patients with immune thrombocytopenias (IT), 17 patients with chronic myelocytic leukemia (CML) and 14 patients with multiple myeloma (MM) was studied by silver staining. The average number of nucleoli in MG of normal donors comprised 21.8 per nucleus with a range from 16.6 to 33.7. It was significantly lower in MG of CML patients, and, on the contrary, it was higher in MG of IT patients. The average number of Ag grains per nucleus reflecting their activity in relation to ribosomal RNA synthesis was found to be the highest (127 +/- 32.1) in MG of IT patients but rather low (43.2 +/- 7.2) in CML patients as compared to those of the control (76.5 +/- 11.1) and MM patients (86.0 +/- 5.6). The differences in the functional state of MG in varying diseases as well as possibilities of using this new approach in hematology have been discussed.


The artificial heart--1986: partial fulfillment of a promise.
While certain critically ill patients who are awaiting cardiac transplantation can be stabilized with either drugs, the intraaortic balloon pump, or a left ventricular assist device, other patients do not respond to these therapies and the use of a safe and effective artificial heart can be lifesaving. For this latter group, the pneumatic heart appears to be suitable, although minor improvements will be welcomed. The percutaneous tubes and bulky drive unit are not great disadvantages to the patient who uses an interim heart and remains in a constant care area. The patient with end-stage heart disease who cannot have a heart transplant has little hope at present. Interestingly, the pneumatic heart that is available now might be an acceptable alternative to death if the incidence of stroke could be overcome. In the future, these patients will be better served by either an electrically powered left ventricular assist pump or a totally artificial heart.


Effects of GnRH antagonist on endometrial protein profiles in the window of implantation.
GnRH antagonists can suppress luteinizing hormone and follicle-stimulating hormone (FSH), with less initial stimulatory effect and lower risk of ovarian hyperstimulation syndrome. The effects of GnRH antagonists on embryonic implantation remain controversial. To evaluate the effects of GnRH antagonists, endometrial tissues were biopsied from 12 women with intracytoplasmic sperm injection treatment, in which four subjects undergoing controlled ovulation stimulation with rFSH and GnRH antagonist, four subjects with a GnRH agonist long protocol, and four natural cycle controls. After iTRAQ quantification analysis, 24 proteins showed differential expression between natural cycle and agonist treatment group and 39 proteins between natural cycle and antagonist treatment group. A total of seven proteins demonstrated differential expression only in antagonist treatment group. Bioinformatic analysis implied these proteins can function in cell processes including angiogenesis, cell proliferation, apoptosis, cell migration, and immune response. Furthermore, GnRH antagonist suppressed the function of GNAS and ANPEP, which were important for endometrial functions. Immunohistochemical staining showed that ANPEP was mainly localized in the human endometrial stroma, while ACO2, CDC5L, and GNAS were mainly localized in the glands. This study could provide insights into the effect of GnRH antagonists on the endometrium, and help optimize the embryo implantation and improve the success rate for GnRH antagonist protocol.


Clinical use of indium-111 labeled blood products.
Following the introduction of In-111 oxine as a label for blood cells by McAffee and Thakur in 1976, these procedures have become increasingly important in the practice of nuclear medicine. Of particular interest are studies involving the use of labeled leukocytes for the detection of focal infection. The clinical utility of labeled platelets is less well developed, although the use of platelets to detect the formation of thrombi in blood vessels and on vascular grafts and prostheses is gaining prominence. This report summarizes the techniques presently employed at the University of Minnesota for the labeling of blood products, and their clinical use. Consideration also is given to the desired expertise and cost factors involved in the labeling of leukocytes and platelets.


Vaccines targeting preS1 domain overcome immune tolerance in hepatitis B virus carrier mice.
Strong tolerance to hepatitis B virus (HBV) surface antigens limits the therapeutic effect of the conventional hepatitis B surface antigen (HBsAg) vaccination in both preclinical animal models and patients with chronic hepatitis B (CHB) infection. In contrast, we observed that clinical CHB patients presented less immune tolerance to the preS1 domain of HBV large surface antigen. To study whether targeting the weak tolerance of the preS1 region could improve therapy gain, we explored vaccination with the long peptide of preS1 domain for HBV virions clearance. Our study showed that this preS1-polypeptide rather than HBsAg vaccination induced robust immune responses in HBV carrier mice. The anti-preS1 rapidly cleared HBV virions in vivo and blocked HBV infection to hepatocytes in vitro. Intriguingly, vaccination of preS1-polypeptide even reduced the tolerized status of HBsAg, opening a therapeutic window for the host to respond to the HBsAg vaccine. A sequential administration of antigenically distinct preS1-polypeptide and HBsAg vaccines in HBV carrier mice could finally induce HBsAg/hepatitis B surface antibody serological conversion and clear chronic HBV infection in carrier mice. These results suggest that preS1 can function as a therapeutic vaccine for the control of CHB. (Hepatology 2017;66:1067-1082).


CHECK (Cohort Hip and Cohort Knee): similarities and differences with the Osteoarthritis Initiative.
To describe the osteoarthritis study population of CHECK (Cohort Hip and Cohort Knee) in comparison with relevant selections of the study population of the Osteoarthritis Initiative (OAI) based on clinical status and radiographic parameters. In The Netherlands a prospective 10-year follow-up study was initiated by the Dutch Arthritis Association on participants with early osteoarthritis-related complaints of hip and/or knee: CHECK. In parallel in the USA an observational 4-year follow-up study, the OAI, was started by the National Institutes of Health, on patients with or at risk of symptomatic knee osteoarthritis. For comparison with CHECK, the entire cohort and a subgroup of individuals excluding those with exclusively hip pain were compared with relevant subpopulations of the OAI. At baseline, CHECK included 1002 participants with in general similar characteristics as described for the OAI. However, significantly fewer individuals in CHECK had radiographic knee osteoarthritis at baseline when compared with the OAI (p<0.001). In contrast, at baseline, the CHECK cohort reported higher scores on pain, stiffness and functional disability (Western Ontario and McMaster osteoarthritis index) when compared with the OAI (all p<0.001). These differences were supported by physical health status in contrast to mental health (Short Form 36/12) was at baseline significantly worse for the CHECK participants (p<0.001). Although both cohorts focus on the early phase of osteoarthritis, they differ significantly with respect to structural (radiographic) and clinical (health status) characteristics, CHECK expectedly representing participants in an even earlier phase of disease.


Anti-obesity and anti-inflammatory effects of synthetic acetic acid vinegar and Nipa vinegar on high-fat-diet-induced obese mice.
Recently, food-based bioactive ingredients, such as vinegar, have been proposed as a potential solution to overcome the global obesity epidemic. Although acetic acid has been identified as the main component in vinegar that contributes to its anti-obesity effect, reports have shown that vinegar produced from different starting materials possess different degrees of bioactivity. This study was performed to compare the anti-obesity and anti-inflammatory effects of synthetic acetic acid vinegar and Nipa vinegar in mice fed a high-fat diet. In this work, mice were fed a high-fat diet for 33 weeks. At the start of week 24, obese mice were orally fed synthetic acetic acid vinegar or Nipa vinegar (0.08 and 2 ml/kg BW) until the end of week 33. Mice fed a standard pellet diet served as a control. Although both synthetic acetic acid vinegar and Nipa vinegar effectively reduced food intake and body weight, a high dose of Nipa vinegar more effectively reduced lipid deposition, improved the serum lipid profile, increased adipokine expression and suppressed inflammation in the obese mice. Thus, a high dose of Nipa vinegar may potentially alleviate obesity by altering the lipid metabolism, inflammation and gut microbe composition in high-fat-diet-induced obese mice.


Evaluation of Amniotic Multipotential Tissue Matrix to Augment Healing of Demineralized Bone Matrix in an Animal Calvarial Model.
Amniotic multipotential tissue matrix (AmnioMTM) is a membrane material derived from placental tissues and rich in growth factors that have been reported to have potential in healing bone. This study hypothesized that demineralized bone matrix (DBM) supplemented with AmnioMTM would accelerate healing and bone formation as compared with DBM alone in a critical size (10 mm) rat calvarial bone defect model. Five DBM grafts and 5 DBM supplemented with AmnioMTM grafts were implanted in a 10-mm critical sized defect in 10 rats (1 implant per rat). After 4 weeks, animals were euthanized and defects evaluated by microCT and histology. There were no statistical differences in microCT data for mineral density, percent bone fill, or bone surface to volume ratios between groups, though the bone surface to volume ratio for the amnio-supplemented group suggested increased osteoid activity as compared with the DBM alone group. Histological data also indicated active osteoid activity and induced bone formation in the center of defects implanted with AmnioMTM supplemented graft as compared with DBM graft alone suggesting some potential osteoinductive potential. However, there was no significant difference at the mean percent of newly mineralized bone in the DBM group defect as compared with the AmnioMTM supplemented graft material. These data suggest that while bone formation was not increased at this early time point, the increased osteoid activity and the induction of new bone in the middle of the defect by the AmnioMTM indicates that further study is needed to assess its potential benefit to bone healing and regeneration.


Bile acid signalling and its role in anxiety disorders.
Anxiety disorder is a prevalent neuropsychiatric disorder that afflicts 7.3%~28.0% of the world's population. Bile acids are synthesized by hepatocytes and modulate metabolism via farnesoid X receptor (FXR), G protein-coupled receptor (TGR5), etc. These effects are not limited to the gastrointestinal tract but also extend to tissues and organs such as the brain, where they regulate emotional centers and nerves. A rise in serum bile acid levels can promote the interaction between central FXR and TGR5 across the blood-brain barrier or activate intestinal FXR and TGR5 to release fibroblast growth factor 19 (FGF19) and glucagon-like peptide-1 (GLP-1), respectively, which in turn, transmit signals to the brain via these indirect pathways. This review aimed to summarize advancements in the metabolism of bile acids and the physiological functions of their receptors in various tissues, with a specific focus on their regulatory roles in brain function. The contribution of bile acids to anxiety via sending signals to the brain via direct or indirect pathways was also discussed. Different bile acid ligands trigger distinct bile acid signaling cascades, producing diverse downstream effects, and these pathways may be involved in anxiety regulation. Future investigations from the perspective of bile acids are anticipated to lead to novel mechanistic insights and potential therapeutic targets for anxiety disorders.


Extracting physics of life at the molecular level: A review of single-molecule data analyses.
Studying individual biomolecules at the single-molecule level has proved very insightful recently. Single-molecule experiments allow us to probe both the equilibrium and nonequilibrium properties as well as make quantitative connections with ensemble experiments and equilibrium thermodynamics. However, it is important to be careful about the analysis of single-molecule data because of the noise present and the lack of theoretical framework for processes far away from equilibrium. Biomolecular motion, whether it is free in solution, on a substrate, or under force, involves thermal fluctuations in varying degrees, which makes the motion noisy. In addition, the noise from the experimental setup makes it even more complex. The details of biologically relevant interactions, conformational dynamics, and activities are hidden in the noisy single-molecule data. As such, extracting biological insights from noisy data is still an active area of research. In this review, we will focus on analyzing both fluorescence-based and force-based single-molecule experiments and gaining biological insights at the single-molecule level. Inherently nonequilibrium nature of biological processes will be highlighted. Simulated trajectories of biomolecular diffusion will be used to compare and validate various analysis techniques.


Discovery of the Lyme Disease Agent.
A detailed first-hand account of the events leading up to the discovery of the Lyme disease agent has been lacking. Nearly 40 years have elapsed since the discovery of the organism that was named Borrelia burgdorferi There are thousands of articles in the scientific and medical literature on this organism and the disease that it causes. In the interval since the organism's discovery, however, misconceptions have arisen regarding not only the disease but the discovery itself. Accordingly, with this paper, we aim to fill in the details of this episode in medical history with a joint introduction, first-person accounts by the two authors, a summary of contemporaneous events, and concluding comments. The history of the discovery of the Lyme disease agent has threads originating in different places in the United States. Studies on Long Island, NY, provided the epidemiological thread of studies on rickettsial diseases and babesiosis, linking the latter with the cutaneous manifestation of Lyme disease, now known as erythema migrans. The Long Island thread intersected Montana's Rocky Mountain Laboratories thread of studies on a relapsing fever Borrelia and its cultivation and expertise in vector biology. This intersection made possible the discovery of the spirochete and its recovery from patients. This paper stresses that what may seem to have been an individual scientific discovery is actually the product of several threads coming together and is attributable to more people than appreciated.


Direct selection for P1-sensitive mutants of enteric bacteria.
A method has been developed to isolate mutants sensitive to coliphage P1 from bacterial genera normally not sensitive to this phage. P1clr100KM was used. This phage is heat inducible and confers kanamycin resistance when present as a prophage (in lysogens). P1-sensitive mutants of Klebsiella, Enterobacter, Citrobacter, and Erwinia have been found. This technique provides a well-known genetic system for the study of many bacterial genera that previously had either no such system or only a marginally useful means of genetic manipulation. It also extends the range of possible intergeneric hybrids that may be constructed and studied.


Mite and cockroach allergens activate protease-activated receptor 2 and delay epidermal permeability barrier recovery.
Protease-activated receptor-2 (PAR-2) is known to be involved in epidermal permeability barrier function homeostasis. PAR-2 activation occurs after barrier disruption and further activation of PAR-2 by activating peptide significantly delays barrier recovery rate. Cockroach and house dust mite allergens, both known to be associated with the development of asthma, allergic rhinitis, and atopic dermatitis, have protease activity, which can activate PAR-2. In this study, we investigated the effects of both allergens on the epidermal barrier function as well as on the epidermal calcium gradient. Both allergens, when topically applied on the barrier-disrupted site, increased protease activities in the epidermis and delayed barrier recovery and lamellar body secretion in murine skin. The topical application of PAR-2-specific antagonist or protease inhibitors normalized the barrier recovery. Cockroach allergens induced intracellular calcium oscillations in cultured human keratinocytes through PAR-2-involved pathway, which was confirmed by desensitization protocol. Abnormal calcium ion distribution after barrier disruption was also observed in allergens-applied skin. These results suggest that allergens with protease activity can influence the epidermal permeability barrier homeostasis through PAR-2 activation and consequent modulation of the calcium ions in skin.


Arterioportal shunt: prevalence in small hemangiomas versus that in hepatocellular carcinomas 3 cm or smaller at two-phase helical CT.
To compare the prevalence of arterioportal (AP) shunting associated with (a) small (< or =3 cm) hemangiomas and (b) hepatocellular carcinomas (HCCs) (< or =3 cm) at two-phase helical computed tomography (CT). Two-phase helical liver CT was performed in 107 patients (61 men, 46 women; age range, 25-73 years; mean, 48.6 years) with 169 small hemangiomas and in 384 patients (292 men, 92 women; age range, 18-82 years; mean, 58.3 years) with 598 HCCs 3 cm or smaller. Diagnosis of HCC was verified with histologic findings (n = 30) or typical imaging and clinical findings (n = 568); that of all hemangiomas was verified with typical imaging and clinical findings. Three radiologists retrospectively reviewed all CT images in consensus. Contrast material-enhanced CT scans were obtained during the hepatic arterial and portal venous phases. AP shunt was considered to be present when wedge-shaped or irregularly shaped homogeneous enhancement peripheral to tumor appeared at hepatic arterial phase CT and isoattenuation or slight hyperattenuation in that area appeared at portal phase CT. The prevalence of AP shunting associated with hemangiomas and that associated with HCCs were compared with multivariate model testing. Speed of lesion enhancement (rapid enhancement, when extent of intratumoral enhancement at hepatic arterial phase CT was >50%; slow enhancement, when extent of intratumoral enhancement was < or =50%) and presence of AP shunt were correlated with chi2 or Fisher exact testing. AP shunts were more frequently found in hemangiomas (36 lesions [21.3%]) than in HCCs (25 lesions [4.2%]) (P <.001). Twenty-four (38%) of the 64 hemangiomas with rapid enhancement had AP shunts, whereas only 12 (11.4%) of the 105 hemangiomas with slow enhancement had AP shunts (P <.001). There was no significant difference between prevalence of AP shunt in the 573 HCCs with rapid enhancement (24 lesions, 4.2%) and that in the 25 HCCs with slow enhancement (one lesion, 4.0%). AP shunts were more frequently seen at two-phase helical CT in small hepatic hemangiomas than in HCCs and thus represent a suggestive but not specific finding of hemangioma. Small hemangiomas with AP shunts tend to show rapid rather than slow enhancement.