What is the Next Step after an Electrodiagnostic Study in Children with Polyneuropathies? Rationale for Laboratory and Other Diagnostic Tests.
The etiology of polyneuropathies varies in the pediatric population, where hereditary or metabolic disorders are far more common than in adults. However, treatable polyneuropathies, also prevalent in these settings, are those to prioritize. Moreover, diagnosing subacute and chronic symptoms in children can be challenging compared to adults. Therefore, selecting the best and most relevant laboratory investigations and paraclinical studies is critical. This taskcan be relatively challenging in countries with limited resources or insurance coverage. This study describe the various types of polyneuropathies found in children and their characteristics and suggest an algorithm for using the best laboratory tests in the context of the Iranian healthcare system.


Molecular basis for the repurposing of histamine H2-receptor antagonist to treat COVID-19.
With the world threatened by a second surge in the number of Coronavirus cases, there is an urgent need for the development of effective treatment for the novel coronavirus (COVID-19). Recently, global attention has turned to preliminary reports on the promising anti-COVID-19 effect of histamine H2-receptor antagonists (H2RAs), most especially Famotidine. Therefore, this study was designed to exploit a possible molecular basis for the efficacy of H2RAs against coronavirus. Molecular docking was performed between four H2RAs, Cimetidine, Famotidine, Nizatidine, Ranitidine, and three non-structural proteins viz. NSP3, NSP7/8 complex, and NSP9. Thereafter, a 100 ns molecular dynamics simulation was carried out with the most outstanding ligands to determine the stability. Thereafter, Famotidine and Cimetidine were subjected to gene target prediction analysis using HitPickV2 and eXpression2Kinases server to determine the possible network of genes associated with their anti-COVID activities. Results obtained from molecular docking showed the superiority of Famotidine and Cimetidine compared to other H2RAs with a higher binding affinity to all selected targets. Molecular dynamic simulation and MMPBSA results revealed that Famotidine as well as Cimetidine bind to non-structural proteins more efficiently with high stability over 100 ns. Results obtained suggest that Famotidine and Cimetidine could be a viable option to treat COVID-19 with a mechanism of action that involves the inhibition of viral replication through the inhibition of non-structural proteins. Therefore, Famotidineand Cimetidine qualify for further study as a potential treatment for COVID-19.


Available volume fraction of macromolecules in the extravascular space of a fibrosarcoma: implications for drug delivery.
Steric exclusion of molecules in the extravascular space of tissues can be quantified by the available volume fraction (K(AV)). Despite its clinical importance, however, there is a paucity of data in the literature regarding the available volume fraction of macromolecules in the extravascular space of tumor tissues. In this study, we quantified K(AV) of inulin, BSA, and dextran molecules of Mr 10,000-2,000,000 in polymer gels and fibrosarcoma tissues. The measurement involved: (a) sectioning of gels or tumor tissues into thin slices (approximately 600 microm) using a Vibratome, (b) ex vivo incubation of the slices in solutions containing fluorescently labeled tracers, and (c) quantification of the equilibrium tracer concentrations in both slices and solutions. We found that K(AV) in gels decreased monotonically when the Mr of dextran was increased from Mr 10,000 to 2,000,000. However, K(AV) in tumor tissues was insensitive to the molecular weight of dextran in the range between Mr 10,000 and 40,000. There was a sharp decrease in K(AV) from 0.28 +/- 0.14 to 0.10 +/- 0.06 when the molecular weight was increased from Mr 40,000 to 70,000. In addition to the molecular weight dependence, K(AV) was heterogeneous in tumors, with intertumoral difference being greater than intratumoral variation. The interstitial fluid space, which was quantified by K(AV) of inulin, was 50% of the total tissue volume. These data indicate that the fraction of the extravascular volume in tumors that is accessible to large therapeutic agents is heterogeneous and depends on the size of agents.


The Magnitude and Determinants of Missed Opportunities for Childhood Vaccination in South Africa.
Missed opportunities for vaccination (MOV) may be among the factors responsible for suboptimal vaccination coverage in South Africa. However, the magnitude and determinants of MOV in the country are not known. Thus, this study seeks to assess the prevalence and determinants of MOV in the country. South Africa is sub-divided into nine administrative provinces. We used nationally representative data from the 2016 South African Demographic and Health Survey. We considered MOV to have occurred if a child aged 12-23 months old had not taken all scheduled basic vaccine doses despite having any of the following contacts with health services: delivery in a health facility; postnatal clinic visit; receipt of vitamin A; and any child-related treatment at a health facility. Multilevel logistic regression was used to determine factors associated with MOV. The national prevalence of MOV among children aged 12-23 months was 40.1%. Children whose mothers attended facility-based antenatal care were considerably less likely to experience MOV than those whose mothers did not attend antenatal care: odds ratio (OR) 0.41, 95% confidence interval (CI) 0.19 to 0.88. Conversely, the independent predictor of an increased MOV among children was residence in either the Gauteng province (OR 2.97, 95% CI 1.29 to 6.81) or Mpumalanga province (OR 2.32, 95%CI 1.04 to 5.18); compared to residence in the Free State province. Our findings suggest a high burden of MOV among children in South Africa and that MOV may be associated with individual and contextual factors. The findings also underscore the need for further exploration of the contextual factors contributing to MOV in South Africa.


Spontaneous ripple formation in MoS(2) monolayers: electronic structure and transport effects.
The spontaneous formation of ripples in molybdenum disulfide (MoS2 ) monolayers is investigated via density functional theory based tight-binding Born-Oppenheimer molecular dynamics. Monolayers with different lengths show spontaneous rippling during the simulations. The density of states reveals a decrease in the bandgap induced by the stretching of the MoS2 units due to ripple formation. Significant quenching in electron conductance was also observed. The ripples in the MoS2 monolayers have an effect on the properties of the material and could impact its application in nanoelectronics.


Nutrition in patients undergoing orthotopic liver transplant.
Thirteen patients with severe liver disease had nutritional assessment in the weeks prior to orthotopic liver transplantation. Parameters measured included height and weight, upper arm anthropometry, delayed cutaneous hypersensitivity, total lymphocyte count, serum levels of albumin and transferrin, and plasma amino acids. Weight, when expressed as a percentage of ideal body weight, was greater than 85%, considered the normal lower limit, in all but two patients. However, mean triceps skinfold and arm muscle circumference were 49 +/- 25 and 78 +/- 9% standard, respectively. Mean serum albumin was 2.7 +/- 0.6 g/dl and although mean serum transferrin level was 184 +/- 86, eight patients had levels less than normal. Seven patients were anergic to Multitest CMI (58%) and 12 patients had depressed total lymphocyte count. All these later measurements in the aggregate support a diagnosis of protein-calorie malnutrition. High preoperative levels of amino acids, especially aspartate, phenylalanine, tyrosine, and methionine, were returned to normal by transplantation. We conclude that protein-calorie malnutrition is common in the group of patients likely to require liver transplant, although individual nutritional assessment parameters may lack sensitivity and specificity in determining nutritional status.


CD4 T Lymphocyte-mediated lung disease: steady state between pathological and tolerogenic immune reactions.
Although considerable evidence indicates a role for CD4(+) T lymphocytes (T cells) in airway inflammation, little data exist regarding the mechanisms underlying the induction and regulation of CD4(+) T cell reactivity to lung-specific antigens. To dissect the immunologic and molecular mechanisms of CD4(+) T cell dysregulation, reactivity to a self-antigen expressed in the lung of mice bearing a major histocompatibility complex class-II-restricted T cell receptor specific for this antigen was studied. Transgenic mice developed a progressive interstitial pneumonitis characterized by massive lymphocytic and plasmacytic infiltration of interalveolar septa, a clinical picture closely resembling some of the interstitial lung diseases. Pulmonary inflammation reached a plateau state in older mice with prominent formation of lymphoid follicles but reduced interstitial infiltration. Extensive immunologic characterization of self-reactive CD4(+) T cells isolated from the inflamed lung suggested the induction of regulatory T cells in the site of inflammation. Moreover, inflammation was accompanied by broad changes in the gene expression pattern toward a profile partially resembling that of activated, but strikingly, also that of regulatory CD4(+) T cells. Together our data provide important insights into functional and molecular alterations being associated with the induction and/or regulation of T cell-mediated pulmonary inflammation.


Bioinformatic Analysis and Integration of Transcriptome and Proteome Results Identify Key Coding and Noncoding Genes Predicting Malignancy in Intraductal Papillary Mucinous Neoplasms of the Pancreas.
Intraductal papillary mucinous neoplasms (IPMNs) are precursor lesions of pancreatic ductal adenocarcinoma (PDAC). IPMNs are generally associated with high risk of developing malignancy and therefore need to be diagnosed and assessed accurately, once detected. Existing diagnostic methods are inadequate, and identification of efficient biomarker capable of detecting high-risk IPMNs is necessitated. Moreover, the mechanism of development of malignancy in IPMNs is also elusive. Gene expression meta-analysis conducted using 12 low-risk IPMN and 23 high-risk IPMN tissue samples. We have also listed all the altered miRNAs and long noncoding RNAs (lncRNAs), identified their target genes, and performed pathway analysis. We further enlisted cyst fluid proteins detected to be altered in high-risk or malignant IPMNs and compared them with fraction of differentially expressed genes secreted into cyst fluid. Our meta-analysis identified 270 upregulated and 161 downregulated genes characteristically altered in high-risk IPMNs. We further identified 61 miRNAs and 14 lncRNAs and their target genes and key pathways contributing towards understanding of the gene regulation during the progression of the disease. Most importantly, we have detected 12 genes altered significantly both in cystic lesions and cyst fluid. Our study reports, for the first time, a meta-analysis identifying key changes in gene expression between low-risk and high-risk IPMNs and also explains the regulatory aspect through construction of a miRNA-lncRNA-mRNA interaction network. The 12-gene-signature could function as potential biomarker in cyst fluid for detection of IPMN with a high risk of developing malignancy.


Emergency Department Management of Pediatric Heart Transplant Recipients: Unique Immunologic and Hemodynamic Challenges.
Since the first heart transplant in 1967, there has been significant progress in this field of cardiac transplantation. Approximately 600 pediatric heart transplants are performed every year worldwide. With the increasing number of pediatric heart transplant patients, and given the few tertiary care pediatric transplant centers, adult and pediatric emergency department (ED) providers are increasingly engaged in the care of pediatric heart transplant recipients in the ED. The aim of this article is to review common ED scenarios pertinent to the pediatric heart transplant patients. There are complications unique to this population, such as rejection, opportunistic infections, and medication side effects, that require special considerations, and it is helpful for the emergency medicine (EM) provider to have knowledge about them. The unique immunological challenges in these patients, including rejection and medication side effects and opportunistic infections, make this population fragile, and the knowledge of these challenges is helpful for EM providers.


Penetrating transorbital foreign body with ocular preservation.
Large penetrating transorbital foreign bodies may initially appear to be of a devastating character to the ocular tissues. However, several reports of such large foreign bodies have proved to spare the eye. A case report of a large wooden foreign body with transorbital penetration into the right frontal lobe is reported. The globe remained intact and was only displaced, with a final visual acuity of 20/40. However, complete ophthalmoplegia and ptosis persisted. A low pressure hydrocephalus ensued following intracranial debridement.


Idiopathic bronchiolitis obliterans organizing pneumonia (idiopathic BOOP) in childhood.
Idiopathic bronchiolitis obliterans organizing pneumonia (idiopathic BOOP) is a clinicopathological entity of unknown origin first described by Epler and colleagues in 1985. We report our findings from a study of two Japanese boys aged 12 and 13 years with idiopathic BOOP. The patients showed no respiratory symptoms nor signs of infection. There was no history of exposure to noxious agents or medications in either case. Laboratory data, including hematological, biochemical, and immunological tests, were normal, and pulmonary function tests showed only a slight nonprogressive decrease from normal predicted values in vital capacity and diffusing capacity. Serial radiographs in both cases showed migration of patchy infiltrates. Fiberoptic bronchoscopy was performed for evaluation of eosinophilic pneumonia, but the bronchoalveolar lavage (BAL) revealed no increase in eosinophils and a decrease in CD4/CD8 ratio in the BAL fluid. Histopathological findings of transbronchial lung biopsy (TBLB) specimens were compatible with a diagnosis of BOOP. As neither case has shown clinical abnormalities apart from migratory patchy infiltrates on chest radiographs, both patients have been observed without drug therapy for 3-4 years.


Neuromuscular fatigue during repeated exhaustive submaximal static contractions of knee extensor muscles in endurance-trained, power-trained and untrained men.
The neural and muscular changes during fatigue produced in repeated submaximal static contractions of knee extensors were measured. Three groups of differently adapted male subjects (power-trained, endurance-trained and untrained, 15 in each) performed the exercise that consisted of 10 trials of submaximal static contractions at the level of 40% of maximal voluntary contraction (MVC) force till exhaustion with the inter-trial rest intervals of 1 min. MVC force, reaction time and patellar reflex time components before and after the fatiguing exercise and following 5, 10 and 15 min of recovery were recorded. Endurance-trained athletes had a significantly longer holding times for all the 10 trials compared with power-trained athletes and untrained subjects. However, no significant differences in static endurance between power-trained athletes and untrained subjects were noted. The fatigue test significantly prolonged the time between onset of electrical and mechanical activity (electromechanical delay) in voluntary and reflex contractions. The electromechanical delay in voluntary contraction condition for power-trained and untrained subjects and in reflex condition for endurance-trained subjects had not recovered 15 min after cessation of exercise. No significant changes in the central component of visual reaction time (premotor time of MVC) and latency of patellar reflex were noted after fatiguing static exercise. It is concluded, that in this type of exercise the fatigue development may be largely owing to muscle contractile failure.


Associations between Antiepileptic Use and Hypogammaglobulinaemia: Findings from a Population-Based Case-Control Study Using Data Linkage.
Increased mortality in epilepsy due to infections (other than pneumonia) has been demonstrated. Small case series of people on antiepileptic drugs (AEDs) have described hypogammaglobulinaemia, which may predispose to infections. It is unclear whether hypogammaglobulinaemia is more frequent in people on AEDs, what AEDs it is associated with, or what clinical impact it has. In this population-based study, we aimed to determine whether AEDs were associated with hypogammaglobulinaemia, which AEDs were associated, and whether the associations may be causal. We conducted an unmatched case-control study using data linkage of routinely collected biochemistry, prescribing, and morbidity datasets in North-East Scotland from 2009-2021. Cases were participants with immunoglobulin levels less than the reference range. Controls were those with normal/high immunoglobulin levels. Logistic regression was used to investigate associations between AED exposure and any hypogammaglobulinaemia, adjusting for age, sex, and comorbidity. We also analysed low IgA, IgM, and IgG separately. We analysed "any AED" exposure and common individual drugs separately. Cumulative exposure data were used to determine whether an exposure-response relationship was present. 18,666 cases and 127,157 controls were identified. Use of any AED was associated with increased risk of hypogammaglobulinaemia (adjusted odds ratio [aOR] 1.20 [95% CI: 1.15-1.25]). Phenytoin use was strongly associated with low IgA (aOR 5.90 [95% CI: 3.04, 10.43]). Carbamazepine and lamotrigine were also associated with low IgA. Apart from topiramate, which was associated with a non-significant decrease in odds of hypogammaglobulinaemia, there was a consistent increase in odds of hypogammaglobulinaemia across most AEDs studied. Panhypogammaglobulinaemia was associated with any AED use, carbamazepine, lamotrigine, gabapentin, and multiple AED use. There was evidence of an exposure-response relationship between any AED use and any hypogammaglobulinaemia, low IgA, and low IgG. Carbamazepine and probably lamotrigine also had an exposure-response relationship with any hypogammaglobulinaemia. AEDs may increase hypogammaglobulinaemia risk. Specific classes of immunoglobulins are differentially affected, and the exposure-response analysis suggests this may be causal. Further work should investigate the clinical impact of these findings. Clinicians should check immunoglobulin levels if unusual or recurrent infections occur in patients treated with AEDs.


Immunocytochemical localization of the sigma(1) receptor in the adult rat central nervous system.
In order to characterize the localization of the sigma(1) receptor in the adult rat central nervous system, a polyclonal antibody was raised against a 20 amino acid peptide, corresponding to the fragment 143-162 of the cloned sigma(1) receptor protein. Throughout the rostrocaudal regions of the central nervous system extending from the olfactory bulb to the spinal cord, intense to moderate immunostaining was found to be associated with: (i) ependymocytes bordering the entire ventricular system, and (ii) neuron-like structures located within the parenchyma. Double fluorescence studies confirmed that, throughout the parenchyma, sigma(1) receptor-immunostaining was essentially associated with neuronal structures immunostained for the neuronal marker betaIII-tubulin. In all rats examined, high levels of immunostaining were always associated with neurons located within specific regions including the granular layer of the olfactory bulb, various hypothalamic nuclei, the septum, the central gray, motor nuclei of the hindbrain and the dorsal horn of the spinal cord. In contrast, only faint immunostaining was associated with neurons located in the caudate-putamen and the cerebellum. Electron microscope studies indicated that sigma(1) receptor immunostaining was mostly associated with neuronal perikarya and dendrites, where it was localized to the limiting plasma membrane, the membrane of mitochondria and of some cisternae of the endoplasmic reticulum. At the level of synaptic contacts, intense immunostaining was associated with postsynaptic structures including the postsynaptic thickening and some polymorphous vesicles, whereas the presynaptic axons were devoid of immunostaining. These data indicate that the sigma(1) receptor antibody prepared here, represents a promising tool for further investigating the role of sigma(1) receptors.


Evaluation of developmental dental anomalies in digital panoramic radiographs in Southeast Iranian Population.
The objective of this study was to evaluate the prevalence of dental developmental anomalies in digital panoramic radiographs of the patients referred to the Zahedan medical imaging center and to evaluate the frequency of anomalies regarding the disorders in shape, position and number in the Southeast of Iran. A total of 1172 panoramic radiographs from 581 males and 586 females aged over 16 years were obtained from the files of the Zahedan medical imaging center between the years of 2014 and 2015. The selected radiographs were evaluated in terms of the anomalies such as dilacerations, taurodontism, supernumerary teeth, congenitally missing teeth, fusion, gemination, tooth impaction, tooth transposition, dens invagination, and peg lateral. Then, the anomalies were compared to each other regarding the frequency of the anomaly type (morphological, positional and numerical). Data were evaluated using descriptive statistics such as frequency and percent, and statistical tests such as X(2) at 0.05 significant level using the Statistical Package for the Social Sciences version 16.5. The prevalence of dental anomaly was 213 (18.17%), which was higher in females (9.90) than male, (8.28), however, this difference was not statistically significant (P > 0.05). The prevalence of dilacerated teeth was 62 (5.29%), taurodontism 63 (5.38%), supernumerary teeth 6 (0.51%), congenitally missing teeth 13 (1.11%), fusion 1 (0.09%), gemination 1 (0.09%), impaction 40 (3.41%), transposition 2 (0.18%), dens invagination 16 (1.37) and peg lateral was 9 (0.77%). The prevalence of morphological anomaly was 152 (71.36%), malposition 42 (19.72%) and numerous anomaly was 19 (8.92%). Dental anomalies are relatively common; although their occurrence is not symptomatic, they can lead to several clinical problems in patients. Detailed clinical and radiographic assessment and counseling during patient visits is a critical factor in assessing the patient's degree of difficulty to help the dentist for better preparedness for the treatment.


Population pharmacokinetic analysis of sorafenib in patients with solid tumours.
Sorafenib is a multikinase inhibitor with activity against B-raf, C-raf, VEGFR2, PDGFRβ and FGFR1. Sorafenib is clinically approved for the treatment of renal cell carcinoma (RCC) and hepatocellular carcinoma (HCC). The pharmacokinetics (PK) of sorafenib are highly variable between subjects. Sorafenib exposure increases less than dose proportionally (likely due to limited solubility). Sorafenib undergoes enterohepatic recycling (EHC). This is the first study to characterize the PK of sorafenib using a model based on sorafenib's known disposition characteristics such as delayed/solubility-limited GI absorption and EHC. The parameterization of the EHC model used a square wave function to describe the gall bladder emptying. This study evaluated the effect of baseline bodyweight, BSA, age, gender, liver function parameters, kidney function parameters and genotype with respect to CYP3A4*1B, CYP3A5*3C, UGT1A9*3 and UGT1A9*5 on sorafenib PK. No clinically important covariates were identified. This model can be used to simulate and explore alternative dosing regimens and to develop exposure-response relationships for sorafenib. To characterize the pharmacokinetics (PK) of sorafenib in patients with solid tumours and to evaluate the possible effects of demographic, clinical and pharmacogenetic (CYP3A4*1B, CYP3A5*3C, UGT1A9*3 and UGT1A9*5) covariates on the disposition of sorafenib. PK were assessed in 111 patients enrolled in five phase I and II clinical trials, where sorafenib 200 or 400 mg was administered twice daily as a single agent or in combination therapy. All patients were genotyped for polymorphisms in metabolic enzymes for sorafenib. Population PK analysis was performed by using nonlinear mixed effects modelling (NONMEM). The final model was validated using visual predictive checks and nonparametric bootstrap analysis. A one compartment model with four transit absorption compartments and enterohepatic circulation (EHC) adequately described sorafenib disposition. Baseline bodyweight was a statistically significant covariate for distributional volume, accounting for 4% of inter-individual variability (IIV). PK model parameter estimates (range) for an 80 kg patient were clearance 8.13 l h(-1) (3.6-22.3 l h(-1) ), volume 213 l (50-1000 l), mean absorption transit time 1.98 h (0.5-13 h), fraction undergoing EHC 50% and average time to gall bladder emptying 6.13 h. Overall, population PK analysis was consistent with known biopharmaceutical/PK characteristics of oral sorafenib. No clinically important PK covariates were identified.


Regulation of MHC class I transport in human dendritic cells and the dendritic-like cell line KG-1.
Dendritic cells (DCs) progress through distinct maturational phases; immature DCs capture Ag while mature DCs are optimized for Ag presentation. Proper control of immunity requires regulated compartmentalization of MHC class II molecules. We report that DCs also regulate MHC class I trafficking throughout maturation. Although mature human DCs express high levels of surface MHC class I, immature DCs exhibit lower surface levels while retaining MHC class I-peptide complexes in the Golgi. A cell line, KG-1, behaves similarly. We confirm the similarity of KG-1 to DCs by demonstrating its capacity to present exogenous Ags in an MHC class I-restricted fashion to CD8(+) T cell hybridomas, a phenomenon called cross-presentation. Biochemical characterization of MHC class I trafficking throughout maturation showed that, in early KG-1 dendritic-like cells, surface arrival of MHC class I-peptide complexes is delayed by their retention in the Golgi. In mature dendritic-like cells, these complexes relocate to the surface and their stability increases, concomitant with up-regulation of costimulatory molecules. Maturation induces qualitative changes in the MHC class I-associated peptide repertoire demonstrated by increased thermostability. The differential processing of MHC class I throughout maturation may prevent premature immune activation while promoting T cell responses in lymph nodes to Ags acquired at sites of inflammation.


Inverse correlation of telomerase activity/proliferation of CD4+ T lymphocytes and disease progression in simian immunodeficiency virus-infected nonhuman primates.
Both increased lymphocyte renewal with subsequent exhaustion of the immune system and impaired T-cell renewal have been put forth to account for CD4+ T-cell depletion and development of AIDS in HIV-1-infected humans and SIV-infected nonhuman primates. In the present study, telomeric terminal restriction fragment length and telomerase activity were used as measures of proliferative activity of T lymphocytes from three nonhuman primate species before and after being infected with SIV. In peripheral blood T cells, our data show both species and T-cell-subset-specific differences in proliferative activity accompanied by different patterns of disease progression. A significant postinfection increase in telomerase/proliferative activity in CD4+ T cells from seropositive sooty mangabeys and from normal progressor rhesus macaques was associated with asymptomatic infection or delayed disease progression, respectively, whereas a decrease in telomerase/proliferative activity detected in CD4+ T cells postinfection from SIVsmmPBj14-infected pigtailed macaques was associated with rapid CD4+ T-cell depletion and disease progression. The levels of telomerase activity observed in CD4+ T cells from peripheral blood closely parallelled those seen in CD4+ T cells in lymph node samples from selected animals. Our data suggest that an increase in proliferative activity of T lymphocytes in vivo may be associated with a favorable course of SIV infection in nonhuman primates.


Thyroid Hormone Levels in Preterm Neonates with Birth Weight Less than 2500 g, Treated with Phenobarbital.
Indicatively, phenobarbital can impair thyroid function in adults and children. The present research aims to evaluate the thyroid hormone levels in preterm neonates who had received phenobarbital treatment. This study was conducted on preterm neonates who weighed less or equal to 2500 g when phenobarbital was prescribed for treatment in the first 15 days of life. TSH and total T4 measurements were performed before and three months after initiation of phenobarbital. In this study, the sum of preterm neonates stood at 94, of which 53 were girls, with a mean birth weight of 2004.41 ± 315.41g. Weight of 8.5% were under 1500 g. The mean gestational age was estimated at 33.64 ± 2.01 weeks. Mean T4 levels were 12.24 ± 1.96 and 12.07 ± 1.95 (p=0.334), and mean TSH levels were 5.34 ± 2.14 and5.15 ± 2.15 (p=0.376) before and after prescribing phenobarbital, respectively. The same results were compared based on sex, gestational age, birth weight, and height for T4 and TSH and T4 based on head circumference. The only significant difference was TSH in preterm infants with head circumference <32 cm before and after prescribing phenobarbital (p=0.030). In preterm newborns that had less than 2500 g birth weight, phenobarbital did not significantly alter the serum thyroid hormone levels.


Ehrenfest modeling of cavity vacuum fluctuations and how to achieve emission from a three-level atom.
A much-needed solution for the efficient modeling of strong coupling between matter and optical cavity modes is offered by mean-field mixed quantum-classical dynamics, where a classical cavity field interacts self-consistently with quantum states of matter through Ehrenfest's theorem. We previously introduced a modified mean-field approach, referred to as decoupled mean-field (DC-MF) dynamics, wherein vacuum fluctuations of the cavity field are decoupled from the quantum-mechanical ground state as a means to resolve an unphysical drawing of energy from the vacuum fluctuations by a two-level atom. Here, we generalize DC-MF dynamics for an arbitrary number of (nondegenerate) atomic levels and show that it resolves an unphysical lack of emission from a three-level atom predicted by conventional mean-field dynamics. We furthermore show DC-MF to provide an improved description of reabsorption and (resonant) two-photon emission processes.


Peritoneal and nodal gliomatosis with endometriosis, accompanied with ovarian immature teratoma: a case study and literature review.
Gliomatosis peritonei (GP) indicates the peritoneal implantation of mature neuroglial tissue and is usually accompanied by ovarian mature or immature teratoma. Here, we report a case of ovarian immature teratoma associated with gliomatosis involving the peritoneum, lymph nodes and Douglas' pouch, where gliomatosis coexisted with endometriosis. As far as we know, only seven cases of GP have been reported as coexisting with endometriosis. Eight cases with mature glial tissue in the lymph nodes, i.e., nodal gliomatosis, have been published either in association with GP or in its absence. Metaplasia of pluripotent coelomic stem cells has been suggested to be responsible for the pathogenesis of endometriosis and GP rather than implantation metastases of ovarian teratomatous tumor with varying maturation. This theory is also applied to GP independently of ovarian teratomatous tumors. To the best of our knowledge, nodal gliomatosis coexisting with GP and also involving endometriosis has not yet been reported.


The human dbl-proto-oncogene product is a cytoplasmic phosphoprotein which is associated with the cytoskeletal matrix.
The translational product of the dbl oncogene is a 66 kDa (p66) protein with no apparent sequence similarity to any of the known oncogene products, whereas the human dbl proto-oncogene encodes a translational product of 115 kDa (p115). We compared proto-dbl p115 and dbl p66 with respect to their subcellular localization, biogenesis and post-translational modifications. Like p66, p115 was found to be a cytoplasmic phosphoprotein present in both cytosol and crude membrane preparations. Membrane fractionation studies revealed that p115 as well as p66 were primarily associated with fractions enriched in plasma membranes, suggesting that this subcellular compartment is a likely site of action of dbl proteins. The membrane-associated forms of p115 and p66 were fairly resistant to solubilization by nonionic detergents, suggesting that dbl proteins associate with the cytoskeletal matrix. p115 was also found to be phosphorylated primarily on serine residues. However, p115 was phosphorylated to a lesser extent as compared to the phosphorylated form of p66. The half-life of proto-dbl p115 was significantly shorter (1 hour) than that of dbl p66 (5-6 h). The higher stability of p66 is likely due to the acquisition of unrelated human sequences and/or to the deletion of the N-terminal region of proto-dbl.


Comparability of activity monitors used in Asian and Western-country studies for assessing free-living sedentary behaviour.
This study aims to compare the outputs of the waist-worn Active style Pro HJA-350IT (ASP; used in studies with Asian populations), the waist-worn ActiGragh™GT3X+ using the normal filter (GT3X+) and the thigh-worn activPAL3 (AP) in assessing adults' sedentary behaviour (total sedentary time, number of breaks) under free-living conditions. Fifty healthy workers wore the three monitors simultaneously during their waking hours on two days, including a work day and a non-work day. Valid data were at least 10 hours of wearing time, and the differences between monitors on the sedentary outputs using the AP as criterion measurement were analyzed by ANOVA. The number of participants who had complete valid data for work day and non-work day was 47 and 44, respectively. Total sedentary time and breaks estimated by the AP were respectively 466.5 ± 146.8 min and 64.3 ± 24.9 times on the work day and 497.7 ± 138.3 min and 44.6 ± 15.4 times on the non-work day. In total sedentary time, the ASP estimated 29.7 min (95%CI = 7.9 to 51.5) significantly shorter than the AP on the work day but showed no significant difference against the AP on the non-work day. The GT3X+ estimated 80.1 min (54.6 to 105.6) and 52.3 (26.4 to 78.2) significantly longer than the AP on the work day and the non-work day, respectively. For the number of breaks from sedentary time, on both days, the ASP and the GT3X+ estimated significantly more than the AP: 14.1 to 15.8 times (6.3 to 22.5) for the ASP and 27.7 to 28.8 times (21.8 to 34.8) for the GT3X+. Compared to the AP as the criterion, the ASP can underestimate total sedentary time and the GT3X+ can overestimate it, and more so at the lower levels of sedentary time. For breaks from sedentary time, compared to the AP, both the GT3X+ the ASP can overestimate.


An iterative method for improved protein structural motif recognition.
We present an iterative algorithm that uses randomness and statistical techniques to improve existing methods for recognizing protein structural motifs. Our algorithm is particularly effective in situations where large numbers of sufficiently diverse examples of the motif are not known. These are precisely the situations that pose significant difficulties for previously known methods. We have implemented our algorithm and we demonstrate its performance on the coiled coil motif. We test our program LearnCoil on the domain of 3-stranded coiled coils and subclasses of 2-stranded coiled coils. We show empirically that for these motifs, our method overcomes the problem of limited data.


Common misperceptions: the press and Victorian views of crime.
After a string of successes in the early nineteenth century, the Victorian movement to reform criminal punishment began to falter. Despite evidence to the contrary, the populace grew convinced that violent crime was on the rise. A frequency analysis of The Times and The Manchester Guardian suggests that this misperception was due to a drastic increase in crime coverage by the periodicals of the day.


Molecular cloning, expression, and analysis of the genes of the homoprotocatechuate catabolic pathway of Escherichia coli C.
The molecular cloning and fine-structure analysis of the homoprotocatechuate (hpc) catabolic pathway genes of Escherichia coli C are described. The genes were located in two operons, hpcBCDEF and hpcGH, that were very closely linked. A regulatory gene, hpcR, involved in the expression of both operons was also identified. Various subclones isolated in the study were useful in the production of chemical intermediates of the pathway. The availability of one such compound facilitated the discovery of a previously unrecognized isomerase involved in the catabolic sequence.


Water chemistry poses health risks as reliance on groundwater increases: A systematic review of hydrogeochemistry research from Ethiopia and Kenya.
Reliance on groundwater is increasing in Sub-Saharan Africa as development programmes work towards improving water access and strengthening resilience to climate change. In lower-income areas, groundwater supplies are typically installed without water quality treatment infrastructure or services. This practice is underpinned by an assumption that untreated groundwater is typically suitable for drinking due to the relative microbiological safety of groundwater compared to surface water; however, chemistry risks are largely disregarded. This article systematically reviews groundwater chemistry results from 160 studies to evaluate potential health risk in two case countries: Ethiopia and Kenya. Most studies evaluated drinking water suitability, focusing on priority parameters (fluoride, arsenic, nitrate, or salinity; 18 %), pollution impacts (10 %), or overall suitability (45 %). The remainder characterised general hydrogeochemistry (13 %), flow dynamics (10 %), or water quality suitability for irrigation (3 %). Only six studies (4 %) reported no exceedance of drinking water quality thresholds. Thus, chemical contaminants occur widely in groundwaters that are used for drinking but are not regularly monitored: 78 % of studies reported exceedance of contaminants that have direct health consequences ranging from hypertension to disrupted cognitive development and degenerative disease, and 81 % reported exceedance of aesthetic parameters that have indirect health impacts by influencing perception and use of groundwater versus surface water. Nevertheless, the spatiotemporal coverage of sampling has substantial gaps and data availability bias is driven by a) the tendency for research to concentrate in areas with known water quality problems, and b) analytical capacity limitations. Improved in-country analytical capacity could bolster more efficient assessment and prioritisation of water chemistry risks. Overall, this review demonstrates that universal and equitable access to safe drinking water (Sustainable Development Goal target 6.1) will not be achieved without wider implementation of groundwater treatment, thus a shift is required in how water systems are designed and managed.


Claudin expression in the rat endolymphatic duct and sac - first insights into regulation of the paracellular barrier by vasopressin.
Hearing and balance functions of the inner ear rely on the homeostasis of the endolymphatic fluid. When disturbed, pathologic endolymphatic hydrops evolves as observed in Menière's disease. The molecular basis of inner ear fluid regulation across the endolymphatic epithelium is largely unknown. In this study we identified the specific expression of the tight junction (TJ) molecules Claudin 3, 4, 6, 7, 8, 10, and 16 in epithelial preparations of the rat inner ear endolymphatic duct (ED) and endolymphatic sac (ES) by high-throughput qPCR and immunofluorescence confocal microscopy. Further we showed that Claudin 4 in the ES is a target of arginine-vasopressin (AVP), a hormone elevated in Menière's disease. Moreover, our transmission-electron microscopy (TEM) analysis revealed that the TJs of the ED were shallow and shorter compared to the TJ of the ES indicating facilitation of a paracellular fluid transport across the ED epithelium. The significant differences in the subcellular localization of the barrier-forming protein Claudin 3 between the ED and ES epithelium further support the TEM observations. Our results indicate a high relevance of Claudin 3 and Claudin 4 as important paracellular barrier molecules in the ED and ES epithelium with potential involvement in the pathophysiology of Menière's disease.


Analytic, Preanalytic, and Clinical Validation of p53 IHC for Detection of TP53 Missense Mutation in Prostate Cancer.
Purpose:TP53 missense mutations may help to identify prostate cancer with lethal potential. Here, we preanalytically, analytically, and clinically validated a robust IHC assay to detect subclonal and focal TP53 missense mutations in prostate cancer.Experimental Design: The p53 IHC assay was performed in a CLIA-accredited laboratory on the Ventana Benchmark immunostaining system. p53 protein nuclear accumulation was defined as any p53 nuclear labeling in >10% of tumor cells. Fifty-four formalin-fixed paraffin embedded (FFPE) cell lines from the NCI-60 panel and 103 FFPE prostate cancer tissues (88 primary adenocarcinomas, 15 metastases) with known TP53 mutation status were studied. DU145 and VCaP xenografts were subjected to varying fixation conditions to investigate the effects of preanalytic variables. Clinical validation was performed in two partially overlapping radical prostatectomy cohorts.Results: p53 nuclear accumulation by IHC was 100% sensitive for detection of TP53 missense mutations in the NCI-60 panel (25/25 missense mutations correctly identified). Lack of p53 nuclear accumulation was 86% (25/29) specific for absence of TP53 missense mutation. In FFPE prostate tumors, the positive predictive value of p53 nuclear accumulation for underlying missense mutation was 84% (38/45), whereas the negative predictive value was 97% (56/58). In a cohort of men who experienced biochemical recurrence after RP, the multivariable HR for metastasis among cases with p53 nuclear accumulation compared with those without was 2.55 (95% confidence interval, 1.1-5.91).Conclusions: IHC is widely available method to assess for the presence of deleterious and heterogeneous TP53 missense mutations in clinical prostate cancer specimens. Clin Cancer Res; 23(16); 4693-703. ©2017 AACR.


[Distribution in the diencephalon of the rat of retrogradely primuline-labelled dopamine-containing neurons forming spinal projections].
Distribution of primuline-labeled dopamine-containing neurons in the rat diencephalon following injection of fluorochrome into upper thoracic spinal cord was studied using a combination of catecholamine fluorescence method and retrograde cell-labeling. It was shown that only neurons of group A11 located in the dorsal hypothalamus and caudal thalamus give rise to the spinal projections. Non-catecholamine cells were also labeled in this region. Group A11 comprised 173 +/- 4 catecholamine-containing neurons. Most of them (up to 86%) were retrogradely labeled with primuline. The number of labeled dopamine-containing neurons increased in the rostro-caudal direction. Possible functional role of the dopaminergic diencephalon-spinal neuronal system is discussed.


Novel mechanisms and treatment approaches in autism spectrum disorder.
Autism Spectrum Disorder (ASD) is a complex neurodevelopmental disorder characterized by marked heterogeneity in biology, expression, and response to treatment. The past decade has yielded considerable progress in understanding the underlying biological mechanisms, in characterizing the earliest behavioral phenotype(s), and in developing and evaluating effective treatments for ASD. This review highlights recent research advances in genetics and neuroimaging, as well as in novel behavioral and psychopharmacological treatment approaches, arguing for the value of trans-disciplinary initiatives to move the field forward exponentially. Despite considerable complexity, patterns are beginning to emerge that can inform the identification of novel treatment targets and approaches. The next generation of major innovations in ASD research will involve collaborations across genetics/genomics, neuroimaging, and intervention science. Such efforts, currently under way, hold tremendous promise for exponentially increasing our capacity to understand the mechanisms that contribute to the emergence of ASD and to develop and evaluate personalized interventions that yield maximal impact in a meaningful way.


Characterization of genetically targeted neuron types in the zebrafish optic tectum.
The optically transparent larval zebrafish is ideally suited for in vivo analyses of neural circuitry controlling visually guided behaviors. However, there is a lack of information regarding specific cell types in the major retinorecipient brain region of the fish, the optic tectum. Here we report the characterization of three previously unidentified tectal cell types that are specifically labeled by dlx5/6 enhancer elements. In vivo laser-scanning microscopy in conjunction with ex vivo array tomography revealed that these neurons differ in their morphologies, synaptic connectivity, and neurotransmitter phenotypes. The first type is an excitatory bistratified periventricular interneuron that forms a dendritic arbor in the retinorecipient stratum fibrosum et griseum superficiale (SFGS) and an axonal arbor in the stratum griseum centrale (SGC). The second type, a GABAergic non-stratified periventricular interneuron, extends a bushy arbor containing both dendrites and axons into the SGC and the deepest sublayers of the SFGS. The third type is a GABAergic periventricular projection neuron that extends a dendritic arbor into the SGC and a long axon to the torus semicircularis, medulla oblongata, and anterior hindbrain. Interestingly, the same axons form en passant synapses within the deepest neuropil layer of the tectum, the stratum album centrale. This approach revealed several novel aspects of tectal circuitry, including: (1) a glutamatergic mode of transmission from the superficial, retinorecipient neuropil layers to the deeper, output layers, (2) the presence of interneurons with mixed dendrite/axon arbors likely involved in local processing, and (3) a heretofore unknown GABAergic tectofugal projection to midbrain and hindbrain. These observations establish a framework for studying the morphological and functional differentiation of neural circuits in the zebrafish visual system.


Entresto, a New Panacea for Heart Failure?
Heart Failure (HF) is one of the main healthcare burdens in the United States and in the world. Many drugs are approved and used in practice for management of this condition; including beta blockers, diuretics, aldosterone antagonists, Angiotensin Converting Enzyme Inhibitors (ACEI's), and Angiotensin Receptor Blockers (ARBs). Recently, the Food and Drug Administration (FDA) approved a drug with brand name Entresto (Sacubitril/Valsartan or LCZ696), an angiotensin receptor neprilysin inhibitor for the use in Heart Failure with Reduced Ejection Fraction (HFrEF) patients instead of ACEI's and ARBs. The drug works through angiotensin receptor blockage via valsartan as well as neprilysin inhibition with sacubitril. This represented a new milestone in managing heart failure patients and provided yet another therapy in our armamentarium. This article reviews the stages that led to the development of this drug, the failure of its preceding agents, the lessons we have learnt, and the current trials of Entresto for new indications.


Secondary sewage treatment versus ocean outfalls: an assessment.
Simplified models have been developed to obtain order of magnitude estimates of the oxygen demand of municipal and industrial wastes and of their potential eutrophication effects in the marine environment. The models have been applied to assess two major corrective actions that might be considered for such pollution problems-secondary sewage treatment and ocean outfalls. It is concluded that the arguments for secondary sewage treatment as the proper corrective action are not compelling and that the problem should be reexamined with appropriate scientific and engineering evaluations. One goal of such scientific evaluations should be a more thorough understanding of the life histories of possible trace contaminants and pathogens which may have long biological, chemical, or geological retention times in the marine environment.


A survey of health-fitness evaluation centers.
Twenty-three medically oriented private health-fitness evaluation centers in the United States were surveyed to determine the types of tests available, protocols used, the availability of exercise prescriptions, qualifications of employees, and facility use. All centers administered an electrocardiogram exercise tolerance test, but great variability existed with regard to the number and type of other tests given and services available. Questions in the survey explored the availability of cardiovascular, pulmonary function, musculoskeletal, body composition, and lifestyle evaluations. Some centers were restricted to testing solely for cardiovascular function, while others were complete wellness centers. The centers had a range of 8 to 325 patrons per month, and in general, they tested more men than women.


NACOB presentation to ASB Young Scientist Award: Postdoctoral. The impact of boundary conditions and mesh size on the accuracy of cancellous bone tissue modulus determination using large-scale finite-element modeling. North American Congress on Biomechanics.
The apparent properties of cancellous bone are determined by a combination of both hard tissue properties and microstructural organization. A method is desired to extract the underlying hard tissue properties from simple mechanical tests, free from the complications of microstructure. It has been suggested that microCT voxel-based large-scale finite element models could be employed to accomplish this goal (van Rietbergen et al., 1995, Journal of Biomechanics, 28, 69-81). This approach has recently been implemented and it is becoming increasingly popular as finite element models increase in size and sophistication (Fyhrie et al., 1997, Proceedings of the 43rd Annual Meeting of the Orthopaedic Research Society, San Francisco, CA, p. 815; van Rietbergen et al., 1997, Proceedings of the 43rd Annual Meeting of the Orthopaedic Research Society, San Francisco, CA, p. 62). However, no direct quantitative measurements of the accuracy of this method applied to porous structures such as cancellous bone have been made. This project demonstrates the feasibility of this approach by quantifying its best-case accuracy in determining the trabecular hard tissue modulus of analogues fabricated of a material with known material properties determined independently by direct testing. In addition we were able to assess the impact of mesh size and boundary conditions on accuracy. We found that the assumption of a frictionless boundary condition in the parallel plate compression loading configuration was a significant source of error that could be overcome with the use of rigid end-caps similar to those used by Keaveny et al. (1997 Journal of Orthopaedic Research, 15(1), 101-110). In conclusion, we found that this approach is an effective method for determining the average trabecular hard tissue properties of human cancellous bone with an expected practical accuracy level better than 5%.


New Strategy for Minimally Invasive Endoscopic Surgery to Treat Infectious Spondylodiscitis in the Thoracolumbar Spine.
Eradicating infection, protecting neurologic function, and maintaining structural alignment are the 3 objectives of treatment for infectious spondylodiscitis. For some patients, surgery may be necessary to achieve these goals; however, open surgeries are associated with high morbidity and mortality in elderly patients and those with multiple comorbidities. Endoscopic surgery provides a minimally invasive surgical option for obtaining a culture sample to aid identification of pathogens, while also providing a route for adequate decompression and drainage. The clinical results of this study were analyzed. To evaluate the efficacy and safety of spinal endoscopic surgery, the basic characteristics of patients analyzed and their inflammatory markers, pain levels, and local kyphotic angles were recorded before surgery and at regular intervals after surgery. The patients' cultured pathogens and previous antibiotic treatments were also recorded and analyzed. Retrospective observational study (institutional review board: CMUH 105-REC2-101). Inpatient surgery center. From October 2006 to March 2017, of 508 patients who received spinal endoscopic surgery, 60 with infectious spondylodiscitis were treated using this new strategy. All 60 patients underwent plain film radiography and enhanced magnetic resonance imaging of the affected region to obtain evidence of infectious spondylodiscitis. The role of a computed tomography-guided biopsy and some indications for open surgery were replaced with endoscopic surgery. All the patients reported rapid pain relief after endoscopic surgery and antibiotic treatment. No significant changes in sagittal alignment were observed in final follow-up radiography images. Causative pathogens were identified in 34 patients (culture rate: 77.27%) without previous antibiotic treatment. The patients' erythrocyte sedimentation rates and C-reactive protein levels had decreased significantly 3 months after endoscopic surgery. Two patients (3.3%) experienced infection relapse following initial endoscopic surgery; both of them were efficiently resolved through a second round of endoscopic surgery. No surgery-related complications were observed and no open spinal surgery was required during the follow-up period. This was a retrospective study; bias was unavoidable because of the single-center nature of the study design. Regarding the culture rate, recurrence rate, kyphotic change, and surgery-related complications, this new strategy for endoscopic surgery is safe and effective for treating infectious spondylodiscitis in the thoracic or lumbar spine and may be considered a new trend in treating diseases of this type. Spine, endoscopic, discectomy, spondylodiscitis, minimally invasive surgery.


MDC1 cleavage by caspase-3: a novel mechanism for inactivating the DNA damage response during apoptosis.
Recently, we identified the "apoptotic ring," containing phosphorylated histone H2AX (γ-H2AX), as an early chromatin modification during apoptosis. Because γ-H2AX initiates the DNA damage response (DDR), we tested whether the apoptotic H2AX response leads to the full recruitment of the DDR factors that normally coordinate DNA repair and cell-cycle checkpoints. We show that the apoptotic H2AX response does not recruit the DDR factors because MDC1 (mediator of DNA damage checkpoint protein 1), which normally binds to γ-H2AX in response to DNA damage and amplifies the DDR, is cleaved by caspase-3. This cleavage separates the BRCT and FHA domains of MDC1 and constitutes a novel mechanism for the inactivation of DNA repair in apoptotic cells. Also, we show that downregulation of MDC1 increases the apoptotic response to TRAIL. Together, these results implicate MDC1 in the cellular apoptotic response.


Resveratrol enhances the anti-tumor activity of the mTOR inhibitor rapamycin in multiple breast cancer cell lines mainly by suppressing rapamycin-induced AKT signaling.
The anti-tumor activity of rapamycin is compromised by the feedback-loop-relevant hyperactive PI3K and ERK-MAPK pathway signaling. In breast cancer cells treated with rapamycin, we observed a moderate increase of AKT phosphorylation (P-AKT) in a rapamycin resistant cell line, MDA-MB-231, as well as a slight increase of P-AKT in a rapamycin sensitive cell line, MCF-7. We found that resveratrol, a natural phytoalexin, suppressed the phosphorylation and activation of the PI3K/AKT pathway in all the three breast cancer cell lines that we tested. It also had a weak inhibitory effect on the activation of the mTOR/p70S6K pathway in two cell lines expressing wildtype PTEN, MCF-7 and MDA-MB-231. The combined use of resveratrol and rapamycin resulted in modest additive inhibitory effects on the growth of breast cancer cells, mainly through suppressing rapamycin-induced AKT activation. We, therefore, reveal a novel combination whereby resveratrol potentiates the growth inhibitory effect of rapamycin, with the added benefit of preventing eventual resistance to rapamycin, likely by suppressing AKT signaling. We also present data herein that PTEN is an important contributor to resveratrol's growth suppressive effects and its potentiation of rapamycin in this therapeutic scenario, as resveratrol's suppression of rapamycin-mediated induction of P-AKT is both PTEN-dependent and -independent. Thus, the resveratrol-rapamycin combination may have therapeutic value in treating breast cancer and perhaps other processes where mTOR is activated.


Double-contrast CT arthrography of the cartilage of the patellofemoral joint.
For assessment of the value of computed tomography (CT) as a method for evaluating the cartilage of the patellofemoral joint, 100 knees were examined in 91 patients. With the patient in a supine position with the knee flexed at a 30 degree angle, a sufficient quantity of air and positive contrast material was required to obtain an accurate CT picture of the cartilage. Normal cartilage images of 53 knees and abnormal images of 47 knees were obtained by use of CT. The latter group was classified into eight types. When the CT findings of 70 knees were compared with arthroscopy and/or arthrotomy, the results showed that CT diagnosis was accurate in 68 of 70 knees, with a coincidence rate of 97.1%. Cartilage thickness at an arbitrary site and the extent of the ulcer within the deep layers were observed easily by CT. Although examination exclusively with arthroscopy had distinct disadvantages (e.g., the inability to assess either the specific thickness of the cartilage or the extent of the ulcers within deep layers), the combined use of arthroscopy with CT provided a precise three-dimensional image of the cartilage. CT was found to be a valuable noninvasive imaging method for patellofemoral joint cartilage evaluation.


Crossing boundaries: a comprehensive survey of medical licensing laws and guidelines regulating the interstate practice of pathology.
In the United States, recent judicial interpretation of interstate licensure laws has found pathologists guilty of malpractice and, more importantly, the criminal practice of medicine without a license. These judgments against pathologists highlight the need for a timely and comprehensive survey of licensure requirements and laws regulating the interstate practice of pathology. For all 50 states, each state medical practice act and state medical board website was reviewed. In addition, each medical board was directly contacted by electronic mail, telephone, or US registered mail for information regarding specific legislation or guidelines related to the interstate practice of pathology. On the basis of this information, states were grouped according to similarities in legislation and medical board regulations. This comprehensive survey has determined that states define the practice of pathology on the basis of the geographic location of the patient at the time of surgery or phlebotomy. The majority of states (n=32) and the District of Columbia allow for a physician with an out-of-state license to perform limited consultation to a physician with the specific state license. Several states (n=5) prohibit physicians from consultation without a license for the specific state. Overall, these results reveal the heterogeneity of licensure requirements between states. Pathologists who either practice in multiple states, send cases to out-of-state consultants, or serve as consultants themselves should familiarize themselves with the medical licensure laws of the states from which they receive or send cases.


Food Hygiene Practices of Rural Women and Microbial Risk for Children: Formative Research in Nepal.
Formative research was conducted in a rural hill setting in Nepal during April-June 2012 to inform the design of an intervention to promote safe food hygiene practices. A variety of methods underpinned by Behavior Centered Design theory and Hazard Analysis Critical Control Points was used to pinpoint key risk behaviors and their environmental and psychological determinants in 68 households with a mother having a child aged 6-59 months. These included video recordings, observation of daily routine, teach-the-researcher sessions, in-depth interviews, observations of actual behaviors, focus group discussions, motive mapping, microbiological assessment, and identification of critical control points. Physical settings, especially the kitchen, form a challenging environment for mothers, including family members in rural hill settings of Nepal to practice adequate food hygiene behaviors. Prevalent food hygiene practices of mothers were inadequate, leading to frequent exposure of young children to highly contaminated food, water, and milk. We identified six critical control points; of these, five needed improving. Determinants of these behaviors included physical and social environment as well as psychological brief and individual motives. Five key food hygiene risk behaviors are suggested for prioritization. While designing a food hygiene intervention package, consideration should be given to the physical, biological, and social environment, and immediate motives behind each practice should be taken into consideration while framing key messages. Creative and engaging activities should be designed around the motives of nurture, disgust, affiliation, and social status/respect.


The effects of phosphorus limitation on carbon metabolism in diatoms.
Phosphorus is an essential element for life, serving as an integral component of nucleic acids, lipids and a diverse range of other metabolites. Concentrations of bioavailable phosphorus are low in many aquatic environments. Microalgae, including diatoms, apply physiological and molecular strategies such as phosphorus scavenging or recycling as well as adjusting cell growth in order to adapt to limiting phosphorus concentrations. Such strategies also involve adjustments of the carbon metabolism. Here, we review the effect of phosphorus limitation on carbon metabolism in diatoms. Two transcriptome studies are analysed in detail, supplemented by other transcriptome, proteome and metabolite data, to gain an overview of different pathways and their responses. Phosphorus, nitrogen and silicon limitation responses are compared, and similarities and differences discussed. We use the current knowledge to propose a suggestive model for the carbon flow in phosphorus-replete and phosphorus-limited diatom cells.This article is part of the themed issue 'The peculiar carbon metabolism in diatoms'.


[Effect of adaptation to cold and physical training on heat emission during muscle work].
People, for a long time exposed to cold and physical training or cold only, exhibit strengthening of thermal regulatory reactions aimed at the preservation of heat in the body in the state of rest, as well as during muscular tension. Adaptation of the organism to cold in the situation of limited motor activity lead to the increase of the thickness of the body "cover" under the conditions of rest in the cold due to the decrease of the temperature of skeletal muscles, while physical training in the cold facilitates stabilization of muscular temperature and, correspondingly, lessening of the thickness of the "cover". At the initial stage of muscular activity thermal regulatory reactions of the organism are aimed at the decrease in heat emission, which promotes accelerated heating of the muscles.


Rifampin resistance mutations in the rpoB gene of Enterococcus faecalis impact host macrophage cytokine production.
Antibiotic-resistant bacteria in the genus Enterococcus are a major cause of nosocomial infections and are an emergent public health concern. Similar to a number of bacterial species, resistance to the antibiotic rifampicin (RifR) in enterococci is associated with mutations in the gene encoding the β subunit of RNA polymerase (rpoB). In Mycobacterium tuberculosis, RifRrpoB mutations alter mycobacterial surface lipid expression and are associated with an altered IL-1 cytokine response in macrophages upon infection. However, it is not clear if RifR mutations modulate host cytokine responses by other bacteria. To address this question, we utilized Enterococcus faecalis (E. faecalis). Here, we treated human monocyte-derived macrophages with heat-inactivated wild type or RifRrpoB mutants of E. faecalis and found that RifR mutations reduced IL-1β cytokine production. However, RifR mutations elicited other potent pro- and anti-inflammatory responses, indicating that they can impact other immune pathways beyond IL-1R1 signaling. Our findings suggest that immunomodulation by mutations in rpoB may be conserved across diverse bacterial species and that subversion of IL-1R1 pathway is shared by RifR bacteria.


Carotid baroreflex regulation of plasma renin levels.
The influence of carotid sinus pressure (CSP) on plasma renin activity (PRA) was studied in vagotomized, alpha-chloralose-anesthetized salt-deprived (<1 meq Na+/day) or salt-loaded (100-125 meq Na+day) cats. Changes in renal vascular resistance were produced by varying the pressure in the isolated perfused carotid sinus while maintaining renal blood flow constant in one kidney. Renal flow-pressure determinations were made at high, medium, and low CSP. Venous blood samples were withdrawn for PRA determination by radioimmunoassay after 10 min at each CSP. Low CSP resulted in an increased renal resistance and a dramatic elevation of the PRA in both groups. High CSP also resulted in an increase in PRA (due to contralateral variable-flow kidney) accompanied by a decrease in renal vascular resistance. No difference was found between PRA during medium CSP ad initial control PRA. Propranolol blocked the increase in PRA during low CSP but had no effect on the elevated PRA during high CSP. It is concluded that low CSP produced an increase in PRA that was associated with a reflex activation of renal sympathetic nerves.


Dynamic Computed Tomography Myocardial Perfusion Imaging: Comparison of Clinical Analysis Methods for the Detection of Vessel-Specific Ischemia.
The clinical analysis of myocardial dynamic computed tomography myocardial perfusion imaging lacks standardization. The objective of this prospective study was to compare different analysis approaches to diagnose ischemia in patients with stable angina referred for invasive coronary angiography. Patients referred for evaluation of stable angina symptoms underwent adenosine-stress dynamic computed tomography myocardial perfusion imaging with a second-generation dual-source scanner. Quantitative perfusion parameters, such as blood flow, were calculated by parametric deconvolution for each myocardial voxel. Initially, perfusion parameters were extracted according to standard 17-segment model of the left ventricle (fully automatic analysis). These were then manually sampled by an operator (semiautomatic analysis). Areas under the receiver-operating characteristic curves of the 2 different approaches were compared. Invasive fractional flow reserve ≤0.80 or diameter stenosis ≥80% on quantitative coronary angiography was used as reference standard to define ischemia. We enrolled 115 patients (88 men; age 57±9 years). There were 72 of 286 (25%) vessels causing ischemia in 52 of 115 (45%) patients. The semiautomatic analysis method was better than the fully automatic method at predicting ischemia (areas under the receiver-operating characteristic curves, 0.87 versus 0.69; P<0.001) with readings obtained in the endocardial myocardium performing better than those in the epicardial myocardium (areas under the receiver-operating characteristic curves, 0.87 versus 0.72; P<0.001). The difference in performance between blood flow, expressed as relative to remote myocardium, and absolute blood flow was not statistically significant (areas under the receiver-operating characteristic curves, 0.90 versus 0.87; P=ns). Endocardial perfusion parameters obtained by semiautomatic analysis of dynamic computed tomography myocardial perfusion imaging may permit robust discrimination between coronary vessels causing ischemia versus not causing ischemia.


Hearing ability in three clownfish species.
Clownfish live in social groups in which there is a size-based dominance hierarchy. In such a context, sonic cues could play a role in social organisation because dominant frequency and pulse length of sounds are strongly correlated with fish size. Data on the hearing ability of these fish are, however, needed to show that they have the sensory ability to detect the frequencies in their sounds. The present study determines the hearing sensitivity in three different anemonefish species (Amphiprion frenatus, Amphiprion ocellaris and Amphiprion clarkii), and compares it with the frequencies in their calls. The frequency range over which the three species can detect sounds was between 75 and 1800 Hz, and they were most sensitive to frequencies below 200 Hz. During sound production, dominant frequency is clearly related (R=0.95) to the fish size, whatever the species. Dominant frequency extends from 370 to 900 Hz for specimens having a size between 55 and 130 mm. The best hearing sensitivity of small specimens were found to be lower than the dominant frequency of their own calls. However, they were found to be close to the dominant frequency of larger fish calls. The interest of juveniles lies in localising the adults and thus their location on the reef.


Stated and revealed inequality aversion in three subject pools.
This paper reports data from three subject pools (n=717 subjects) using techniques based on those of Loewenstein, et al. (1989) and Blanco, et al. (2011) to obtain parameters, respectively, of stated and revealed inequality aversion. We provide a replication opportunity for those papers, with two innovations: (i) a design which allows stated and revealed preferences to be compared at the individual level; (ii) assessment of robustness of findings across subjects from a UK university, a Turkish university and Amazon Mechanical Turk. Our findings on stated aversion to inequality are qualitatively similar to those of Loewenstein, et al. in each of our subject pools, whereas there are notable differences between some of our findings on revealed preference and those of Blanco, et al. We find that revealed advantageous inequality aversion is often stronger than revealed dis-advantageous inequality aversion. In most subject pools, we find some (weak) correlation between corresponding parameters of stated and revealed inequality aversion.


Assessment of a specifically developed bullet casing gun for the stunning of water buffaloes.
Water buffaloes and cattle differ considerably with respect to the anatomy of the head. As a result, captive bolt stunners often fail to reliably produce adequate loss of consciousness in water buffaloes and, thus, do not fulfill animal welfare requirements. The goal of the present study was to assess and validate a new stunning device for water buffaloes meeting animal welfare and occupational safety requirements. The newly designed bullet casing gun uses .357Mag/10.2g hollow point bullets and has additional safety features. Its effectiveness and usability were assessed under practical conditions in an abattoir as based on widely accepted criteria. Stunning resulted in deep unconsciousness in 19 out of 20 water buffaloes. One 9-year old male did not immediately collapse. Except for very old bulls, the device presented herewith provides a means to stun water buffaloes of both sexes effectively and reliably while keeping occupational hazards to a minimum.


Characterization of AMPylation on threonine, serine, and tyrosine using mass spectrometry.
Recent studies have suggested that adenosine 5'-monophosphate (AMP) post-translational modification of proteins could represent a novel molecular signaling pathway. Mass spectrometric fragmentation characteristics of this modification have not previously been described and studied systematically. In this work, we therefore examined the fragmentation pattern of chemically synthesized peptides containing AMPylated Thr, Ser, and Tyr. The formation of characteristic ions and the influence of collision energy (CE) on the detection of characteristic ions and their relative peak intensity are reported. When peptide with AMPylated Ser/Thr underwent collision induced dissociation (CID), peaks at m/z 348.1, 136.1, and 250.1, fragments with AMP group attached, and fragments consistent with neutral loss of 347 Da were major characteristic ions; fragments consistent with neutral loss of 135 Da or 249 Da were weaker and not always detectable. The observations for Tyr AMPylation followed the same general patterns as those for Ser/Thr modification, with the exception that the ions detected for Tyr AMPylation did not include either the peak at m/z 348.1, or fragments with a mass shift of -347 Da. The results described in this paper highlight a series of diagnostic ions, which can be used not only to confidently identify the AMPylation site based on MS and MS/MS data, but also to selectively scan AMPylated peptides in complex protein mixtures.


Congenital Chagas disease.
Two different histological types of congenital Chagas disease are defined. In one type, parasites were seen within the skeletal and cardiac fibers, and in the other, they are found mostly within the cells of the reticuloendothelial system. The latter was often associated with parasitized giant-cells with a single, lobulated, hyperchromatic nucleus. In 24 placentas, the most consistent findings were villous and intervillous inflammatory infiltrates. Amastigotes of Trypanosoma cruzi were found mostly in the chorionic villi and in the chorionic plate, and less frequently in the fetal membranes.


Characterization of parainfluenza type 3 virus isolated from the lung of a lamb with pneumonia.
A virus designated DH-1 was isolated from the lung of a lamb that died during the course of an epizootic of acute respiratory tract disease. The virus contained ribonucleic acid and was sensitive to heat (56 C), lipid solvent, and acid (pH 3.0). Inoculated cell cultures absorbed and culture fluids agglutinated guinea pig erythrocytes. Morphogenic and morphologic characteristics of the isolate were compatible with those of the genus Paramyxovirus of the family Paramyxoviridae. Serologic results indicated that the virus was indistinguishable from prototype parainfluenza virus type 3.


Cardiopulmonary bypass parameters improve the prediction of 30-day mortality following cardiac surgery.
Currently 30-day mortality is commonly used as a quality indicator for cardiac surgery; however, prediction models have not included the role of cardiopulmonary bypass (CPB). We hypothesized that reproducing currently utilised prediction model methods of 30-day mortality using the Australian and New Zealand Collaborative Perfusion Registry (ANZCPR) would identify relevant CPB predictors. Nine centers in Australia and New Zealand collected data using the ANZCPR between 2011-2020. CPB parameter selection was determined by evaluating association with 30-day mortality. Data were divided into model creation (n = 15,073) and validation sets (n = 15,072). Bootstrap sampling and automated variable selection methods were used to develop candidate models. The final model was selected using prediction mean square error and Bayesian Information Criteria. The average receiver operating characteristic (ROC), p-value for Hosmer-Lemeshow chi-squared test and MSE were obtained from multifold validation. In total, 30,145 patients were included, of which 735 (2.4%) died within 30 day of surgery. The area under the ROC curve for the model including CPB parameters was significantly greater than preoperative risk factors only (0.829 vs 0.783, p < 0.001). CPB parameters included in the predictive model were CPB time, red blood cell transfusion, mean arterial pressure <50 mmHg, minimum oxygen delivery, cardiac index <1.6 L/min/m2. CPB parameters improve the prediction of 30-day mortality. Randomised trials designed to evaluate modifiable CPB parameters will determine their impact on mortality.


Hemimasticatory and hemifacial spasm: a common pathophysiology?
The case is described of a 60-year-old woman with a 4-year history of left hemimasticatory spasm. Detailed electrophysiological studies are compared with those from 6 patients with typical idiopathic hemifacial spasm. On the basis of comparable patterns of EMG activity in the 2 conditions, it is concluded that they may have both a common pathophysiology and a peripheral site of involvement.


Dural ectasia of the optic nerve sheath: is it always benign?
A 36-year-old woman with a 3-month history of progressive visual loss had papilledema, dilatation of the optic nerve sheaths and normal cerebrospinal fluid pressures. She was diagnosed as dural ectasia of the optic nerve sheaths and surgical decompression was performed. In this case report, severe visual loss is described as a serious complication of this rare disease and the importance of early surgical intervention is emphasized.


Common and rare variants of DAOA in bipolar disorder.
The D-amino acid oxidase activator (DAOA, previously known as G72) gene, mapped on 13q33, has been reported to be genetically associated with bipolar disorder (BP) in several populations. The consistency of associated variants is unclear and rare variants in exons of the DAOA gene have not been investigated in psychiatric diseases. We employed a conditional linkage method-STatistical Explanation for Positional Cloning (STEPC) to evaluate whether any associated single nucleotide polymorphisms (SNPs) account for the evidence of linkage in a pedigree series that previously has been linked to marker D13S779 at 13q33. We also performed an association study in a sample of 376 Caucasian BP parent-proband trios by genotyping 38 common SNPs in the gene region. Besides, we resequenced coding regions and flanking intronic sequences of DAOA in 555 Caucasian unrelated BP patients and 564 mentally healthy controls, to identify putative functional rare variants that may contribute to disease. One SNP rs1935058 could "explain" the linkage signal in the family sample set (P = 0.055) using STEPC analysis. No significant allelic association was detected in an association study by genotyping 38 common SNPs in 376 Caucasian BP trios. Resequencing identified 53 SNPs, of which 46 were novel SNPs. There was no significant excess of rare variants in cases relative to controls. Our results suggest that DAOA does not have a major effect on BP susceptibility. However, DAOA may contribute to bipolar susceptibility in some specific families as evidenced by the STEPC analysis.


CGRP-dependent signalling pathways involved in mouse models of GTN- cilostazol- and levcromakalim-induced migraine.
Knowledge of exact signalling events during migraine attacks is lacking. Various substances are known to trigger migraine attacks in patients and calcitonin gene-related peptide antagonising drugs are effective against migraine pain. Here, we investigated the signalling pathways involved in three different mouse models of provoked migraine and relate them to calcitonin gene-related peptide and other migraine-relevant targets. In vivo mouse models of glyceryl trinitrate-, cilostazol- and levcromakalim-induced migraine were applied utilising tactile sensitivity to von Frey filaments as measuring readout. Signalling pathways involved in the three models were dissected by use of specific knockout mice and chemical inhibitors. In vivo results were supported by ex vivo wire myograph experiments measuring arterial dilatory responses and ex vivo calcitonin gene-related peptide release from trigeminal ganglion and trigeminal nucleus caudalis from mice. Glyceryl trinitrate-induced hypersensitivity was dependent on both prostaglandins and transient receptor potential cation channel, subfamily A, member 1, whereas cilostazol- and levcromakalim-induced hypersensitivity were independent of both. All three migraine triggers activated calcitonin gene-related peptide signalling, as both receptor antagonism and antibody neutralisation of calcitonin gene-related peptide were effective inhibitors of hypersensitivity in all three models. Stimulation of trigeminal ganglia and brain stem tissue samples with cilostazol and levcromakalim did not result in release of calcitonin gene-related peptide, and vasodilation following levcromakalim stimulation was independent of CGRP receptor antagonism. The mouse models of glyceryl trinitrate-, cilostazol- and levcromakalim- induced migraine all involve calcitonin gene-related peptide signalling in a complex interplay between different cell/tissue types. These models are useful in the study of migraine mechanisms.


Mouse Mesenchyme forkhead 2 (Mf2): expression, DNA binding and induction by sonic hedgehog during somitogenesis.
Cloning and sequencing of mouse Mf2 (mesoderm/mesenchyme forkhead 2) cDNAs revealed an open reading frame encoding a putative protein of 492 amino acids which, after in vitro translation, binds to a DNA consensus sequence. Mf2 is expressed at high levels in the ventral region of newly formed somites, in sclerotomal derivatives, in lateral plate and cephalic mesoderm and in the first and second branchial arches. Other regions of mesodermal expression include the developing tongue, meninges, nose, whiskers, kidney, genital tubercule and limb joints. In the nervous system Mf2 is transcribed in restricted regions of the mid- and forebrain. In several tissues, including the early somite, Mf2 is expressed in cell populations adjacent to regions expressing sonic hedgehog (Shh) and in explant cultures of presomitic mesoderm Mf2 is induced by Shh secreted by COS cells. These results suggest that Mf2, like other murine forkhead genes, has multiple roles in embryogenesis, possibly mediating the response of cells to signaling molecules such as SHH.


Occurrence of Rabl-like telomere clustering in the holocentric chromosomes of the peach potato aphid Myzus persicae (Hemiptera; Aphididae).
Several studies demonstrated that chromosome anchoring to nuclear structures is involved in the organization of the interphase nucleus. The Rabl configuration, a well-studied chromosome organization in the interphase nucleus, has been deeply studied in organisms with monocentric chromosomes but just slightly touched in species with holocentric chromosomes. In the present paper, by means of the isolation and chromosomal mapping of the C0t DNA fraction and chromatin immunoprecipitation with anti-LEM-2 antibodies, we evidenced the presence of few foci where telomeres and subtelomeric regions cluster in the aphid interphase nuclei, suggesting the occurrence of a Rabl-like chromosome configuration. The same experimental approaches also evidenced that most of the repetitive DNA of the 2 X chromosomes is located at the periphery of the nucleus, whereas the ribosomal genes, located at 1 telomere of each X chromosome, are present towards the inner portion of the nucleus, favoring their transcriptional activity.


Adenoviral gene therapy with catalase suppresses experimental optic neuritis.
To determine if adenoviral-mediated transfer of the gene for catalase (CAT), the reactive oxygen species scavenger, suppresses experimental optic neuritis. Gene therapy with CAT delivered by an adeno-associated viral vector was previously shown to suppress experimental optic neuritis. Because the transduction of protein expression with recombinant adeno-associated viral vector is relatively slow, taking weeks to reach full levels, we studied the effects of replication-deficient adenovirus containing CAT in suppressing experimental optic neuritis. Transduction with adenovirus occurs within days of inoculation, thus, it may be more applicable for the treatment of patients with acute optic neuritis. Replication-deficient adenovirus containing CAT was injected above the right optic nerve heads of SJL/J mice that were simultaneously sensitized for experimental allergic encephalomyelitis. For controls, the left eyes were injected with the replication-deficient adenovirus without CAT or no virus. The histological effects of CAT on the lesions of experimental allergic encephalomyelitis were measured by computerized analysis of the myelin sheath area (for demyelination), optic disc area (for optic nerve head swelling), the extent of the cellular infiltrate, extravasated serum albumin labeled with immunogold (for disruption of the blood-brain barrier), and the in vivo hydrogen peroxide reaction product. After 1 month, cell-specific catalase activity, evaluated by the quantitation of catalase immunogold, was increased about 2-fold each in endothelia, oligodendroglia, astrocytes, and axons of the CAT-inoculated right optic nerves compared with the control left optic nerves. The increased cellular levels of catalase reduced demyelination by 30%, optic nerve head swelling by 25%, cellular infiltration by 26%, disruption of the blood-brain barrier by 61%, and in vivo levels of hydrogen peroxide by 81%. Adenoviral-mediated gene transfer increased catalase levels in all optic nerve cell types, and it persisted for 1 month after inoculation. The increased cellular levels of catalase suppressed demyelination and blood-brain barrier disruption at the foci in the optic nerve where prior magnetic resonance imaging and histopathologic studies have demonstrated the demyelinating inflammation of experimental and human optic neuritis. Together, they suggest that gene therapy with CAT may be helpful in the treatment of patients with optic neuritis.


KNOX homeodomain protein directly suppresses the expression of a gibberellin biosynthetic gene in the tobacco shoot apical meristem.
To identify genes targeted by the tobacco KNOX homeodomain protein, Nicotiana tabacum homeobox 15 (NTH15), we have generated an inducible system using the human glucocorticoid receptor. In this system, steroid treatment strictly induced NTH15 function and immediately suppressed the expression of a gibberellin (GA) biosynthetic gene encoding GA 20-oxidase (Ntc12) and also resulted in a decrease in bioactive GA levels. The repression of Ntc12 was observed even when indirect effects were blocked by cycloheximide. NTH15 mRNA was present in corpus cells of the shoot apical meristem (SAM), whereas Ntc12 mRNA was observed in leaf primordia and rib meristem but not in the corpus. Recombinant NTH15 protein strongly bound to a 5-bp dyadsymmetric sequence, GTGAC, in the first intron of Ntc12 in vitro. Mutation of this sequence in the Ntc12 gene abolished the NTH15-dependent suppression of Ntc12 in the corpus of the SAM. Our results indicate that NTH15 directly represses Ntc12 expression in the corpus of the wild-type SAM to maintain the indeterminate state of corpus cells. The suppression of NTH15 within cells at the flanks of the SAM permits GA biosynthesis, which promotes organized cell proliferation and consequently induces the determination of cell fate.


A proteomic survey of rat cerebral cortical synaptosomes.
Previous findings from our laboratory and others indicate that two-dimensional gel electrophoresis (2-DE) can be used to study protein expression in defined brain regions, but mainly the proteins which are present in high abundance in glia are readily detected. The current study was undertaken to determine the protein profile in a synaptosomal subcellular fraction isolated from the cerebral cortex of the rat. Both 2-DE and liquid chromatography - tandem mass spectrometry (LC-MS/MS) procedures were used to isolate and identify proteins in the synaptosomal fraction and accordingly >900 proteins were detected using 2-DE; the 167 most intense gel spots were isolated and identified with matrix-assisted laser desorption/ionization - time of flight peptide mass fingerprinting or LC-MS/MS. In addition, over 200 proteins were separated and identified with the LC-MS/MS "shotgun proteomics" technique, some in post-translationally modified form. The following classes of proteins associated with synaptic function were detected: (a) proteins involved in synaptic vesicle trafficking-docking (e.g., SNAP-25, synapsin I and II, synaptotagmin I, II, and V, VAMP-2, syntaxin 1A and 1B, etc.); (b) proteins that function as transporters or receptors (e.g., excitatory amino acid transporters 1 and 2, GABA transporter 1); (c) proteins that are associated with the synaptic plasma membrane (e.g., post-synaptic density-95/synapse-associated protein-90 complex, neuromodulin (GAP-43), voltage-dependent anion-selective channel protein (VDACs), sodium-potassium ATPase subunits, alpha 2 spectrin, septin 7, etc.); and (d) proteins that mediate intracellular signaling cascades that modulate synaptic function (e.g., calmodulin, calcium-calmodulin-dependent protein kinase subunits, etc.). Other identified proteins are associated with mitochondrial or general cytosolic function. Of the two proteins identified as endoplasmic reticular, both interact with the synaptic SNARE complex to regulate vesicle trafficking. Taken together, these results suggest that the integrity of the synaptosomes was maintained during the isolation procedure and that this subcellular fractionation technique enables the enrichment of proteins associated with synaptic function. The results also suggest that this experimental approach can be used to study the differential expression of multiple proteins involved in alterations of synaptic function.


Management of hypersensitivity reactions to anti-D immunoglobulin preparations.
RhD immunoglobulin G (anti-D) administered to pregnant Rh(-) women prevents Rh isoimmunization. Its use has significantly reduced the incidence of haemolytic disease of the foetus and newborn previously responsible for one death in every 2200 births. In pregnancy, acute drug-induced hypersensitivity reactions including anaphylaxis can have serious deleterious effects on the mother and foetus/neonate. Women can be erroneously labelled as drug allergic as the investigation of hypersensitivity reactions in pregnancy is complex and drug challenges are usually contraindicated. We present three cases of suspected anti-D hypersensitivity clinically presenting as anaphylaxis and delayed transfusion-related reaction. We also propose a new algorithm for the investigations of such reaction. It relies on detailed history, cautious interpretation of skin tests, foetal Rh genotyping from maternal blood and, in some cases, anti-D challenges. This is not to deprive women of anti-D which might put their future pregnancies at risk.


[Prolonged epidural anesthesia in the treatment of frostbite].
A comparative analysis of efficiency of prolonged epidural anesthesia with lidocain in a complex of medical measures has been presented in 91 patients with cold damage of the lower extremities (9 of 3-4 degrees). Application of the method of prolonged epidural anesthesia by lidocain in a combined treatment of frost-bites has allowed to improve clinical course of the disease and hospital stay of the patients 2.5 times, and in 100% of cases to avoid invalidating surgery.


Enhancing the textural and rheological properties of fermentation-induced pea protein emulsion gels with transglutaminase.
The aim of this study was to assess how transglutaminase (TG) impacts the microstructure, texture, and rheological properties of fermentation-induced pea protein emulsion gels. Additionally, the study examined the influence of storage time on the functional properties of these gels. Fermentation-induced pea protein gels were produced in the presence or absence of TG and stored for 1, 4, 8, 12, and 16 weeks. Texture analysis, rheological measurements, moisture content and microstructure evaluation with confocal laser scanning microscopy (CLSM) and 3D image analysis were conducted to explore the effects of TG on the structural and rheological properties of the fermented samples. The porosity of the protein networks in the pea gels decreased in the presence of TG, the storage modulus increased and the textural characteristics were significantly improved, resulting in harder and more springy gels. The gel porosity increased in gels with and without TG after storage but the effect of storage on textural and rheological properties was limited, indicating limited structural rearrangement once the fermentation-induced pea protein emulsion gels are formed. Greater coalescence was observed for oil droplets within the gel matrix after 16 weeks of storage in the absence of TG, consistent with these protein structures being weaker than the more structurally stable TG-treated gels. This study shows that TG treatment is a powerful tool to enhance the textural and rheological properties of fermentation-induced pea protein emulsion gels.


Generalization of the dynamic clamp concept in neurophysiology and behavior.
The idea of closed-loop interaction in in vitro and in vivo electrophysiology has been successfully implemented in the dynamic clamp concept strongly impacting the research of membrane and synaptic properties of neurons. In this paper we show that this concept can be easily generalized to build other kinds of closed-loop protocols beyond (or in addition to) electrical stimulation and recording in neurophysiology and behavioral studies for neuroethology. In particular, we illustrate three different examples of goal-driven real-time closed-loop interactions with drug microinjectors, mechanical devices and video event driven stimulation. Modern activity-dependent stimulation protocols can be used to reveal dynamics (otherwise hidden under traditional stimulation techniques), achieve control of natural and pathological states, induce learning, bridge between disparate levels of analysis and for a further automation of experiments. We argue that closed-loop interaction calls for novel real time analysis, prediction and control tools and a new perspective for designing stimulus-response experiments, which can have a large impact in neuroscience research.


Fluorescent protein barrel fluctuations and oxygen diffusion pathways in mCherry.
Fluorescent proteins (FPs) are valuable tools as biochemical markers for studying cellular processes. Red fluorescent proteins (RFPs) are highly desirable for in vivo applications because they absorb and emit light in the red region of the spectrum where cellular autofluorescence is low. The naturally occurring fluorescent proteins with emission peaks in this region of the spectrum occur in dimeric or tetrameric forms. The development of mutant monomeric variants of RFPs has resulted in several novel FPs known as mFruits. Though oxygen is required for maturation of the chromophore, it is known that photobleaching of FPs is oxygen sensitive, and oxygen-free conditions result in improved photostabilities. Therefore, understanding oxygen diffusion pathways in FPs is important for both photostabilites and maturation of the chromophores. In this paper, we use molecular dynamics calculations to investigate the protein barrel fluctuations in mCherry, which is one of the most useful monomeric mFruit variant. We employ implicit ligand sampling to determine oxygen pathways from the bulk solvent into the mCherry chromophore in the interior of the protein. We also show that these pathways can be blocked or altered and barrel fluctuations can be reduced by strategic amino acid substitutions.


Systemic Convergent Multitarget Interactions of Plant Polyphenols Revealed by Affinity-Based Protein Profiling of Bone Cells Using C-Glucosidic Vescal(ag)in-Bearing Chemoproteomic Probes.
The ellagitannins vescalagin and vescalin, known as actin-dependent inhibitors of osteoclastic bone resorption, were mounted onto chemical probes to explore their interactions with bone cell proteins by means of affinity-based chemoproteomics and bioinformatics. The chemical reactivity of the pyrogallol units of these polyphenols toward oxidation into electrophilic ortho-quinones was exploited using NaIO4 to promote the covalent capture of target proteins, notably those expressed at lower abundance and those interacting with polyphenols at low-to-moderate levels of affinity. Different assays revealed the multitarget nature of both ellagitannins, with 100-370 statistically significant proteins captured by their corresponding probes. A much higher number of proteins were captured from osteoclasts than from osteoblasts. Bioinformatic analyses unveiled a preference for the capture of proteins having phosphorylated ligands and GTPase regulators and enabled the identification of 33 potential target proteins with systemic relevance to osteoclast differentiation and activity, as well as to the regulation of actin dynamics.