Risk factors and genetic bases: the case of attention deficit hyperactivity disorder.
Attention deficit hyperactivity disorder (ADHD) is considered to be the most frequent mental disorder in childhood. Although its diagnosis in the most utilized handbook of psychiatry in the world today - the Diagnostic and statistical handbook of mental disorders (DSM-5) - is based on behaviors of inattention, hyperactivity and impulsivity, numerous attempts to describe the biological bases of the disorder can be found, to be used for and also as risk markers. In this paper, we will critically analyze the validity of studies associated with the search for genetic markers of ADHD. First, a characterization of ADHD by the DSM-5 handbook is presented. Subsequently, the link between ADHD, risk factors and genetic markers is developed. Finally, some conclusions are presented which highlight simplifications and omissions that could have significant consequences.


Peripheral endotoxin increases splenic sympathetic nerve activity via central prostaglandin synthesis.
We tested whether prostaglandin synthesis mediates the lipopolysaccharide (LPS)-induced increase in splenic sympathetic nerve activity. Sprague-Dawley rats were pretreated with intravenous or intracerebroventricular injections of indomethacin, and splenic nerve activity was recorded after intravenous injections of LPS. In vehicle-pretreated rats, 100 micrograms LPS induced a 62.8 +/- 5.6% increase in splenic nerve activity beginning 22.7 +/- 2.7 min postinjection. All vehicle-pretreated animals responded to high (100 micrograms, 5 of 5 animals) and low (10 micrograms, 8 of 8 animals) doses of LPS. Both intravenous (15 mg/kg) and intracerebroventricular (50 micrograms) pretreatments with indomethacin delayed (F1.19 = 30.66, P < 0.001) the increase in nerve activity after 100 micrograms LPS. When given intravenously, 50 micrograms indomethacin (the intracerebroventricular dose) did not delay the response to intravenous LPS, indicating that the effects of intracerebroventricular indomethacin pretreatment were restricted to the central nervous system. Importantly, intracerebroventricular indomethacin reduced (2 of 7 animals) or completely blocked (5 of 7 animals) the splenic nerve response to the low dose of LPS (10 micrograms, iv). The indomethacin effects could not be accounted for by central release of vasopressin because intracerebroventricular injection of indomethacin did not alter baseline nerve activity or blood pressure, whereas intracerebroventricular injection of vasopressin rapidly increased both measures. Additionally, central injection of LPS did not elevate splenic nerve activity, whereas intracerebroventricular injection of prostaglandin E2 induced a rapid (2.2 +/- 2.7 min) increase in splenic nerve activity. These data indicate that central prostaglandin synthesis is an intermediate step whereby systemic LPS elicits an increase in sympathetic outflow to an immune organ.


Sodium-glucose cotransporter-2 inhibitors improve clinical outcomes in patients with type 2 diabetes mellitus undergoing anthracycline-containing chemotherapy: an emulated target trial using nationwide cohort data in South Korea.
Novel hypoglycemic agents, sodium-glucose cotransporter 2 inhibitors (SGLT2i), have shown protective effects against anthracycline (AC)-induced cardiotoxicity and exhibit partial anticancer effects in animal models. However, clinical evidence for this is scarce. This study aimed to evaluate whether SGLT2i improve the clinical outcomes of patients with type 2 diabetes mellitus (T2DM) undergoing AC-containing chemotherapy. A total of 81,572 patients who underwent AC chemotherapy between 2014 and 2021 were recruited from a nationwide Korean cohort. Patients were classified into three groups: patients with T2DM taking SGLT2i (n = 780) and other hypoglycemic agents excluding SGLT2i (non-SGLT2i; n = 3,455) during AC chemotherapy, and the non-DM group (n = 77,337). The clinical outcome was a composite of heart failure hospitalization, acute myocardial infarction, ischemic stroke, and death. After propensity score matching, 779 SGLT2i users were compared with 7800 non-DM patients and 2,337 non-SGLT2i users. The SGLT2i group had better composite outcomes compared with the non-DM group (adjusted hazard ratio [HR] = 0.35, 95% confidence interval [95% CI] = 0.25-0.51) and compared with the non-SGLT2i group (adjusted HR = 0.47, 95% CI = 0.32-0.69). In conclusion, SGLT2i may contribute to improving clinical outcomes in patients with T2DM undergoing AC-containing chemotherapy, through an emulated target trial using Korean nationwide cohort data.


Isolation of a bovine Theileria.
Dual infections of Anaplasma marginale and a Theileria, resembling Theileria mutans, occurred in splenectomized calves inoculated with pooled blood samples from eastern Texas cattle. Theileria was obtained in pure form by treating dually infected cattle with selectively eliminated Anaplasma. These theileria infections were responsible for mild, transient reductions in packed red blood cell volume (PCV).


Ancient pathogen DNA in human teeth and petrous bones.
Recent ancient DNA (aDNA) studies of human pathogens have provided invaluable insights into their evolutionary history and prevalence in space and time. Most of these studies were based on DNA extracted from teeth or postcranial bones. In contrast, no pathogen DNA has been reported from the petrous bone which has become the most desired skeletal element in ancient DNA research due to its high endogenous DNA content. To compare the potential for pathogenic aDNA retrieval from teeth and petrous bones, we sampled these elements from five ancient skeletons, previously shown to be carrying Yersinia pestis. Based on shotgun sequencing data, four of these five plague victims showed clearly detectable levels of Y. pestis DNA in the teeth, whereas all the petrous bones failed to produce Y. pestis DNA above baseline levels. A broader comparative metagenomic analysis of teeth and petrous bones from 10 historical skeletons corroborated these results, showing a much higher microbial diversity in teeth than petrous bones, including pathogenic and oral microbial taxa. Our results imply that although petrous bones are highly valuable for ancient genomic analyses as an excellent source of endogenous DNA, the metagenomic potential of these dense skeletal elements is highly limited. This trade-off must be considered when designing the sampling strategy for an aDNA project.


What role is there for antithymocyte globulin in allogeneic nonmyeloablative canine hematopoietic cell transplantation?
We investigated whether pretransplantation immunosuppression with canine-specific rabbit antithymocyte globulin (ATG), combined with a suboptimal dose of 1 Gy of total body irradiation (TBI), would permit engraftment of canine dog leukocyte antigen-identical marrow. Cumulative ATG doses of 2 to 5 mg/kg produced a T-cell depletion of 1 log in the peripheral blood and 50% in the lymph nodes. Serum levels of ATG peaked on days 4 to 6 after initiation of therapy and became undetectable by day 13 as a result of canine antibody responses to ATG. ATG prolonged allogeneic skin graft survival to 14 days (n = 5), compared with 8 days in control dogs (P = .0003). Five dogs were given marrow transplants after ATG (3.5-5 mg/kg) and 1 Gy of TBI. Posttransplantation immunosuppression consisted of mycophenolate mofetil and cyclosporine. All dogs showed initial engraftment, with maximum donor chimerism levels of 25%. However, only 1 dog achieved sustained engraftment, and 4 rejected their grafts. The duration of engraftment ranged from 8 to > or = 36 weeks (median, 11 weeks), and this is comparable to that in 6 historical controls not given ATG (range, 3-12 weeks; median, 10 weeks; P = .20). The total nucleated cell doses in the marrow grafts had the highest correlation coefficient with the duration of engraftment: 0.82 (P = .09). We concluded that administering ATG before an otherwise suboptimal conditioning dose of 1 Gy of TBI failed to secure uniform stable hematopoietic engraftment.


Theca lutein ovarian cyst may be pathognomonic for trophoblastic neoplasia.
Two cases are presented in which theca lutein tissue was found at the time of indicated pelvic surgery performed long after treatment for hydatidiform mole. This indicator for the presence of trophoblastic tissue was not recognized, and clinical metastatic gestational trophoblastic neoplasia subsequently developed in each patient. Theca lutein tissue found at pelvic surgery merits investigation by beta-human chorionic gonadotropin radioimmunoassay and is likely to be associated with subclinical latent trophoblastic neoplasia.


Mutation in the signal-transducing chain of the interferon-gamma receptor and susceptibility to mycobacterial infection.
IFN-gamma is critical in the immune response to mycobacterial infections, and deficits in IFN-gamma production and response have been associated with disseminated nontuberculous mycobacterial infections. Mutations in the IFN-gamma receptor ligand-binding chain (IFNgammaR1) have been shown to confer susceptibility to severe infection with nontuberculous mycobacteria. However, mutations in the IFN-gamma receptor signal-transducing chain (IFNgammaR2) have not been described. We describe a child with disseminated Mycobacterium fortuitum and M. avium complex infections and absent IFN-gamma signaling due to a mutation in the extracellular domain of IFNgammaR2. In vitro cytokine production by patient PBMCs showed 75% less PHA-induced IFN-gamma production than in normal cells, while patient PHA-induced TNF-alpha production was normal. The normal augmentation of TNF-alpha production when IFN-gamma was added to endotoxin was absent from patient cells. Expression of IFNgammaR1 was normal, but there was no phosphorylation of Stat1 in response to IFN-gamma stimulation. DNA sequence analysis of the gene for IFNgammaR2 showed a homozygous dinucleotide deletion at nucleotides 278 and 279, resulting in a premature stop codon in the protein extracellular domain. This novel gene defect associated with disseminated nontuberculous mycobacterial infection emphasizes the critical role that IFN-gamma plays in host defense against mycobacteria.


Effective management of peristomal pyoderma gangrenosum.
Peristomal pyoderma gangrenosum (PPG) is a skin disorder which manifests as painful exudating, ulcerative lesions. It is generally associated with inflammatory bowel disease (IBD) when affecting patients with a stoma. Patients with PPG typically present to the stoma care nurse (SCN) in the first instance, when the discharging ulcers cause pain, prevent effective stoma appliance application and result in leakage. Anecdotal evidence suggests that clinical nurse specialist posts, such as that of the SCN, are being diluted within some trusts, with a demand for staff to work on wards to cover clinical shifts. As a consequence there is a possibility that these patients will not receive a prompt diagnosis and treatment plan. This article will examine the role of the SCN in managing the patient with PPG, including the use of effective topical treatments. The importance of early diagnosis to prevent escalation of PPG and subsequent prolonged patient discomfort-and the SCN's crucial role in this-is discussed.


[Principles of signal coding by the discharge pattern of neural networks].
Neuronal populations are shown to form rings of stochastic dependence for optimal coding of information. Such a population can only exist in strictly defined stochastic states determined by the number of the stochastic dependence rings. A system consisting of a certain number of neurons, was shown to be able to code and transfer a number of messages equal to square of the number of stochastic states permitted for the given system. The number of messages differing from each other either by the number of characters or their order in the word, would be equal to the number of permitted stochastic states, whereas the number of messages containing the same number of characters in the word would be equal to the difference between the number of permitted stochastic states and the number of neurons in the system. The alphabet consists of two letters: "U" - determination of the stochastic dependence ring; "R" - disruption of this ring and punctuation marks - there are neither determination nor disruption of the stochastic dependence rings.


Effects of anterior cingulate cortex lesions on a continuous performance task for mice.
Important tools in the study of prefrontal cortical-dependent executive functions are cross-species behavioural tasks with translational validity. A widely used test of executive function and attention in humans is the continuous performance task (CPT). Optimal performance in variations of this task is associated with activity along the medial wall of the prefrontal cortex, including the anterior cingulate cortex (ACC), for its essential components such as response control, target detection and processing of false alarm errors. We assess the validity of a recently developed rodent touchscreen continuous performance task (rCPT) that is analogous to typical human CPT procedures. Here we evaluate the performance of mice with quinolinic acid-induced lesions centred on the ACC in the rCPT following a range of task parameter manipulations designed to challenge attention and impulse control. Lesioned mice showed a disinhibited response profile expressed as a decreased response criterion and increased false alarm rates. ACC lesions also resulted in a milder increase in inter-trial interval responses ('ITI touches') and hit rate. Lesions did not affect discriminative sensitivity d'. The disinhibited behaviour of ACC lesioned animals was stable and not affected by the manipulation of variable task parameter manipulations designed to increase task difficulty. The results are in general agreement with human studies implicating the ACC in the processing of inappropriate responses. We conclude that the rCPT may be useful for studying prefrontal cortex function in mice and has the capability of providing meaningful links between animal and human cognitive tasks.


Agreement between spouses and children in descriptions of personality change in Alzheimer's disease.
The purpose of the present study was to evaluate the agreement among spouses and children in their describing the current and past personality of patients with Alzheimer's disease (AD) and to determine the relations between their descriptions and selected demographic and clinical variables. The subjects were 22 dementia out-patients who fulfilled the DSM-III-R criteria for uncomplicated dementia of the Alzheimer's type and the NINCDS-ADRDA criteria for AD. Spouses and children of these patients were separately administered Brooks and McKinlay's personality inventory consisting of 18 pairs of adjectives that characterize the extremes of a behavioral dimension, and were asked to judge where the subject's demeanor fell on a five-point scale, varying from +2 to -2, in which zero was regarded as neutral. Spouses and children reported marked changes in most measured behavioral dimensions following the onset of AD. Spouses identified significant changes on 14/18 items and children on 13/18 items. Spouses and children agreed on practically all items concerning personality attributes before the onset of illness, and on 16/18 items after it. Changes in personality were not correlated with the studied demographic characteristics of patients, spouses and children, nor with the cognitive deficits and illness duration of the patients.


An Unexpected Route to an Essential Cofactor: Escherichia coli Relies on Threonine for Thiamine Biosynthesis.
Metabolism consists of biochemical reactions that are combined to generate a robust metabolic network that can respond to perturbations and also adapt to changing environmental conditions. Escherichia coli and Salmonella enterica are closely related enterobacteria that share metabolic components, pathway structures, and regulatory strategies. The synthesis of thiamine in S. enterica has been used to define a node of the metabolic network by analyzing alternative inputs to thiamine synthesis from diverse metabolic pathways. To assess the conservation of metabolic networks in organisms with highly conserved components, metabolic contributions to thiamine synthesis in E. coli were investigated. Unexpectedly, we found that, unlike S. enterica, E. coli does not use the phosphoribosylpyrophosphate (PRPP) amidotransferase (PurF) as the primary enzyme for synthesis of phosphoribosylamine (PRA). In fact, our data showed that up to 50% of the PRA used by E. coli to make thiamine requires the activities of threonine dehydratase (IlvA) and anthranilate synthase component II (TrpD). Significantly, the IlvA- and TrpD-dependent pathway to PRA functions in S. enterica only in the absence of a functional reactive intermediate deaminase (RidA) enzyme, bringing into focus how these closely related bacteria have distinct metabolic networks. In most bacteria, including Salmonella strains and Escherichia coli, synthesis of the pyrimidine moiety of the essential coenzyme, thiamine pyrophosphate (TPP), shares enzymes with the purine biosynthetic pathway. Phosphoribosylpyrophosphate amidotransferase, encoded by the purF gene, generates phosphoribosylamine (PRA) and is considered the first enzyme in the biosynthesis of purines and the pyrimidine moiety of TPP. We show here that, unlike Salmonella, E. coli synthesizes significant thiamine from PRA derived from threonine using enzymes from the isoleucine and tryptophan biosynthetic pathways. These data show that two closely related organisms can have distinct metabolic network structures despite having similar enzyme components, thus emphasizing caveats associated with predicting metabolic potential from genome content.


Assessing or predicting adoption of telehealth using the diffusion of innovations theory: a practical example from a rural program in New Mexico.
In New Mexico, a large rural state, it was anticipated that telehealth would bring significant value to health care delivery, improve local capacity for patient care, decrease the need for patient travel, diminish professional isolation, provide an avenue for enhanced professional education and information sharing, increase access to care, and ultimately improve health status. During the course of an evaluation of the University of New Mexico Center for Telehealth's rural telemedicine program, we used a grounded theory approach to assess barriers to the adoption of telemedicine and components of successful adoption. We then turned to the diffusion of innovations theory to better understand the dynamic interactions between the characteristics of telehealth and the social system in which it is applied. In doing so, we learned that the type of innovation decision involved in the adoption of telehealth appears to be particularly important in determining adoption. In this article we demonstrate that diffusion theory can be a useful framework for evaluating telehealth programs. We also suggest that the development of a predictive tool for prospective assessment would be useful, and could be applied when new telehealth programs are being planned.


Platelet-derived growth factor (PDGF) receptor-alpha activates c-Jun NH2-terminal kinase-1 and antagonizes PDGF receptor-beta -induced phenotypic transformation.
Platelet-derived growth factor (PDGF) is a potent mitogen for mesenchymal cells. The PDGF B-chain (c-sis proto-oncogene) homodimer (PDGF BB) and v-sis, its viral counterpart, activate both alpha- and beta-receptor subunits (alpha-PDGFR and beta-PDGFR) and mediate anchorage-independent growth in NIH3T3 cells. In contrast, the PDGF A chain homodimer (PDGF AA) activates alpha-PDGFR only and fails to induce phenotypic transformation. In the present study, we investigated alpha- and beta-PDGFR specific signaling pathways that are responsible for the differences between the transforming ability of PDGF AA and BB. To study PDGF BB activation of beta-PDGFR, we established NIH3T3 clones in which alpha-PDGFR signaling is inhibited by a dominant-negative alpha-PDGFR, or an antisense construct of alpha-PDGFR. Here, we demonstrate that beta-PDGFR activation alone is sufficient for PDGF BB-mediated anchorage-independent cell growth. More importantly, inhibition of alpha-PDGFR signaling enhanced PDGF BB-mediated phenotypic transformation, suggesting that alpha-PDGFR antagonizes beta-PDGFR-induced transformation. While both alpha- and beta-receptors effectively activate ERKs, alpha-PDGFR, but not beta-PDGFR, activates stress-activated protein kinase-1/c-Jun NH(2)-terminal kinase-1 (JNK-1). Inhibition of JNK-1 activity using a dominant-negative JNK-1 mutant markedly enhanced PDGF BB-mediated anchorage-independent cell growth, demonstrating an antagonistic role for JNK-1 in PDGF-induced transformation. Consistently, overexpression of wild-type JNK-1 reduced PDGF BB-mediated transformation. Taken together, the present study showed that alpha- and beta-PDGFRs differentially regulate Ras-mitogen-activated protein kinase pathways critical for regulation of cell transformation, and transformation suppressing activity of alpha-PDGFR involves JNK-1 activation.


HLA-linked nonresponsiveness to Cryptomeria japonica pollen antigen. I. Nonresponsiveness is mediated by antigen-specific suppressor T cell.
The objective of this study was to elucidate the cellular mechanism of IgE nonresponse to the Cryptomeria japonica (Japanese cedar) pollen antigen (CPAg), which was shown in our previous study to be HLA-linked (1). We established an assay system for the measurement of small amounts of anti-CPAg IgE antibody, both in an antigen-specific and isotype-specific manner, and a culture system to induce antigen-driven IgE antibody synthesis in vitro. By using these methods, we clarified that the function of the HLA-DR molecule in the CPAg-driven IgE response is similar to that of I-A or I-E molecule in mice, namely the product of immune response genes (Ir-genes), because anti-HLA-DR monoclonal antibody blocked the response, and the interaction between monocyte and monocyte-depleted peripheral blood lymphocytes (PBL) to respond to CPAg was restricted by HLA-DR. Furthermore, PBL from nonresponders revealed a specific IgE response to CPAg when the Leu-2+3- T cell fraction was depleted, thereby suggesting that even nonresponders have Leu-2-3+ T cell and B cell clones specific for CPAg, and they apparently show no response due to the presence of CPAg-specific Leu-2+3- suppressor T cells. This suppressor T cell fraction abolished the IgE response of the autologous B + monocyte + Leu-2-3+ T cell in a CPAg-specific manner. The current cellular analysis together with our previous genetic analysis strongly suggest that the HLA-linked IgE nonresponse to CPAg is mediated by CPAg-specific suppressor T cells. The HLA-linked gene controlling the nonresponsiveness to CPAg is thus designated as the immune suppression gene for CPAg (Is-CPAg). Mapping of Is-CPAg within HLA-DQ subregion is discussed.


Multi-omic prediction of incident type 2 diabetes.
The identification of people who are at high risk of developing type 2 diabetes is a key part of population-level prevention strategies. Previous studies have evaluated the predictive utility of omics measurements, such as metabolites, proteins or polygenic scores, but have considered these separately. The improvement that combined omics biomarkers can provide over and above current clinical standard models is unclear. The aim of this study was to test the predictive performance of genome, proteome, metabolome and clinical biomarkers when added to established clinical prediction models for type 2 diabetes. We developed sparse interpretable prediction models in a prospective, nested type 2 diabetes case-cohort study (N=1105, incident type 2 diabetes cases=375) with 10,792 person-years of follow-up, selecting from 5759 features across the genome, proteome, metabolome and clinical biomarkers using least absolute shrinkage and selection operator (LASSO) regression. We compared the predictive performance of omics-derived predictors with a clinical model including the variables from the Cambridge Diabetes Risk Score and HbA1c. Among single omics prediction models that did not include clinical risk factors, the top ten proteins alone achieved the highest performance (concordance index [C index]=0.82 [95% CI 0.75, 0.88]), suggesting the proteome as the most informative single omic layer in the absence of clinical information. However, the largest improvement in prediction of type 2 diabetes incidence over and above the clinical model was achieved by the top ten features across several omic layers (C index=0.87 [95% CI 0.82, 0.92], Δ C index=0.05, p=0.045). This improvement by the top ten omic features was also evident in individuals with HbA1c <42 mmol/mol (6.0%), the threshold for prediabetes (C index=0.84 [95% CI 0.77, 0.90], Δ C index=0.07, p=0.03), the group in whom prediction would be most useful since they are not targeted for preventative interventions by current clinical guidelines. In this subgroup, the type 2 diabetes polygenic risk score was the major contributor to the improvement in prediction, and achieved a comparable improvement in performance when added onto the clinical model alone (C index=0.83 [95% CI 0.75, 0.90], Δ C index=0.06, p=0.002). However, compared with those with prediabetes, individuals at high polygenic risk in this group had only around half the absolute risk for type 2 diabetes over a 20 year period. Omic approaches provided marginal improvements in prediction of incident type 2 diabetes. However, while a polygenic risk score does improve prediction in people with an HbA1c in the normoglycaemic range, the group in whom prediction would be most useful, even individuals with a high polygenic burden in that subgroup had a low absolute type 2 diabetes risk. This suggests a limited feasibility of implementing targeted population-based genetic screening for preventative interventions.


Polymorphisms in tumour necrosis factor-alpha, transforming growth factor-beta, interleukin-10, interleukin-6, interferon-gamma, and outcome of hepatitis C virus infection.
Cytokines play a key role in the regulation of immune responses. In hepatitis C virus infection (HCV), the production of inappropriate cytokine levels appears to contribute to viral persistence and to affect response to therapy. Cytokine genes are polymorphic at specific sites, and certain mutations located within coding/regulatory regions have been shown to affect the overall expression and secretion of cytokines. The aim of this study was to investigate the frequency of genotypes associated with polymorphisms of TNF-alpha, TGF-beta, IL-10, IL-6, and IFN-gamma and to determine their association with the outcome of HCV infection. Genotyping was carried out by polymerase chain reaction sequence-specific primers on genomic DNA isolated from 158 individuals. Of these, 66 had spontaneously recovered from infection (persistently HCV RNA negative), while 92 had persistent infection (persistently HCV RNA positive). All patients were genotyped as high or low producers of TNF-alpha and IL-6 and high, intermediate, or low producers of TGF-beta, IL-10, and IFN-gamma based on single nucleotide substitutions. A significant proportion of patients with viral clearance were genotyped with a low IL-6 production profile, whereas those with persistent infection were genotyped with a high production profile (P = 0.02). No associations were observed between polymorphisms of TNF-alpha, IL-10, or IFN-gamma and viral clearance or persistent infection. Furthermore, there were no associations between cytokine genotypes and severity of disease. Inheritance of some genotypes associated with polymorphisms of cytokine genes, such as IL-6, may be host genetic factors associated with outcome of HCV in a well-defined ethnically homogeneous cohort.


The aquatic vertebrate embryo as a sentinel for toxins: zebrafish embryo dechorionation and perivitelline space microinjection.
Pollution of aquatic ecosystems poses a serious threat to aquatic organisms and ultimately the entire ecosystem. Understanding how a toxin affects embryonic development is key to determining the risk a pollutant represents to the environment. Extraembryonic membranes, such as the chorion of fish eggs, provide a protective barrier between the embryo and the environment. Although the fish chorion excludes many chemical pollutants, some noxious agents can still gain access to the aquatic embryo. Therefore a monitoring system that tests the effects directly upon the embryo must be established. Although exposure to a single toxin in the laboratory can determine the concentration at which a pollutant becomes a health or environmental hazard, embryos and adults in nature are not merely affected by a single chemical, but are exposed to mixtures of different pollutants. Zebrafish (Danio rerio) and medaka (Oryzias latipes) embryos were employed for the rapid observation of the effects of single chemicals and chemical mixtures on development. Using dechorionation and a perivitelline space microinjection system, the embryos were effective sentinels for low concentrations of aquatic pollutants. The developmental effects of small quantities of toxins were observed. Embryos treated during the late gastrula stage of development with hexachlorobenzene (HCB); 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD); toluene; benzene; or mixtures of these chemicals developed cardiovascular abnormalities. The zebrafish dechorionation exposure technique, Micro Intrachorionic Zebrafish Embryo Live Laboratory test, was especially effective in testing the pollutant mixtures. Combinations of both TCDD and benzene (as well as the toluene and benzene combinations) were tested and the mixtures acted synergistically; the combinations were more toxic than either chemical by itself. Hexachlorobenzene- and TCDD-treated embryos tested positively for expression of cytochrome P450 1A indicating that the cytochrome metabolic pathways were already functional in these early embryos, and suggested that a product of the cytochrome system may be involved in HCB and TCDD pollution associated cardiovascular defects.


Translesion synthesis DNA polymerase η exhibits a specific RNA extension activity and a transcription-associated function.
Polymerase eta (Polη) is a low fidelity translesion synthesis DNA polymerase that rescues damage-stalled replication by inserting deoxy-ribonucleotides opposite DNA damage sites resulting in error-free or mutagenic damage bypass. In this study we identify a new specific RNA extension activity of Polη of Saccharomyces cerevisiae. We show that Polη is able to extend RNA primers in the presence of ribonucleotides (rNTPs), and that these reactions are an order of magnitude more efficient than the misinsertion of rNTPs into DNA. Moreover, during RNA extension Polη performs error-free bypass of the 8-oxoguanine and thymine dimer DNA lesions, though with a 103 and 102-fold lower efficiency, respectively, than it synthesizes opposite undamaged nucleotides. Furthermore, in vivo experiments demonstrate that the transcription of several genes is affected by the lack of Polη, and that Polη is enriched over actively transcribed regions. Moreover, inactivation of its polymerase activity causes similar transcription inhibition as the absence of Polη. In summary, these results suggest that the new RNA synthetic activity of Polη can have in vivo relevance.


On the molecular-replacement problem in the presence of merohedral twinning: structure of the N-terminal half-molecule of human lactoferrin.
The structure of a hemihedrally twinned protein crystal with two molecules in the asymmetric unit was solved by molecular replacement. The protein, a site-specific mutant of the N-terminal half-molecule of human lactoferrin, is able to undergo an internal rigid-body domain motion. Therefore, determining the structure required the independent positioning of four protein domains. The molecular-replacement solutions were obtained using a conventional real-space rotation function, and a translation function based on the linear correlation coefficient. Once the molecules were positioned, it was necessary to assign them to the appropriate twin domain. Several methods for doing this are described, one of which leads to a determination of the volume of each twin domain. In the appendix to the paper we discuss the interpretation of the self-rotation function in the presence of merohedral twinning.


Internal jugular veins out flow in patients with multiple sclerosis:a catheter venography study.
To investigate an examiner-independent catheter venography protocol that could be used to reliably diagnose venous outflow abnormalities in patients with multiple sclerosis (MS) and chronic cerebrospinal venous insufficiency and to determine whether venous angioplasty is effective in the treatment of these abnormalities. A total of 313 patients with MS and 12 patients with end-stage renal disease underwent echo-color Doppler sonography and catheter venography of the internal jugular veins (IJVs) to evaluate contrast medium clearance time. In patients with venous outflow anomalies, balloon angioplasty of the IJVs was performed. A contrast medium clearance time cutoff value of 4 seconds or less provided the maximal combination of sensitivity and specificity for the right IJV (sensitivity, 73.4%; specificity, 100%) and left IJV (sensitivity, 91.4%; specificity, 100%). IJVs with a clearance time between 4.1 and 6 seconds had moderate delayed flow (MDF), and IJVs with a clearance time longer than 6 seconds had severe delayed flow (SDF); 89% of patients showed MDF/SDF through at least one IJV, 79% showed MDF/SDF through both IJVs, and only 5% showed normal flow in both IJVs. Balloon angioplasty was immediately able to improve flow in at least one IJV in 69% of patients, but venous flow was normalized in both veins in only 37% of patients; SDF persisted after angioplasty in 32% of patients. There is a high prevalence of abnormal delayed flow through IJVs in patients with MS. Venous angioplasty was effective in only a minority of patients with SDF.


[Characteristics of the ultrastructural organization of endothelial cells in cardiac and omental blood capillaries based on cryofractographic data].
Using cryofractography, endotheliocytes of heart and omentum blood capillaries were studied, different patterns of distribution of membranous particles being observed of the "P" and "E" faces of the fracture plans of endothelial cell cytomembranes. Three types of distribution of micropinocytotic vesicles can be distinguished: the linear, hexagonal, and the clusteral type. The number of micropinocytotic vesicles was sharply reduced near the region of intercellular contacts. On the fracture plan, attached pinocytotic vesicles look like "craters" on the "E" face, and like circumvallatue papillae on the "P" face. Cell membranes, making intercellular contacts, have a network structure consisting of ridges on the "P" face and of grooves on the "E" face. Such a molecular membrane organization is discussed from the standpoint of modern concepts of the receptor--cytosceletal complex. It is supposed that this complex may control processes of transendothelial mass--transfer.


Lipid metabolism and regulation of membrane trafficking.
The past 20 years have witnessed tremendous progress in our understanding of the molecular machinery that controls protein and membrane transport between organelles (Scheckman R, Orci L. Coat proteins and vesicle budding. Science 1996;271: 1526-1533 and Rothman JE. Mechanisms of intracellular protein transport. Nature 1994;372: 55-63.) The research efforts responsible for these impressive advances have largely focused on the identification and characterization of protein factors that participate in membrane trafficking events. The role of membranes and their lipid constituents has received considerably less attention. Indeed, until rather recently, popular models for mechanisms of membrane trafficking had relegated membrane lipids to the status of a passive platform, subject to deformation by the action of coat proteins whose polymerization and depolymerization govern vesicle budding and fusion reactions. The 1990s, and particularly its last half, has brought fundamental reappraisals of the interface of lipids and lipid metabolism in regulating intracellular membrane trafficking events. Some of the emerging themes are reviewed here.


[ETHIOLOGY OF LEPTOSPIROSIS IN GEORGIA].
Investigation of leptospirosis was initiated in 1950s, when the first cases in humans were detected in Gudauta rayon. Serology test confirmed the diagnose. High level of antibodies to serogroup Pomona was revealed in all cases. Later on the infection encountered as sporadic cases, the exception was 1986, when a large outbreak was reported in Sukhumi. At present leptospirosis is widely spread in the country, especially in Ajara region, where indicators of morbidity recently reached 11.4 per 100 thousand population. It is likely that warm humid climate creates exceptionally favorite conditions for spreading of Leptospira. In order to determine the true magnitude of the infection in the country a seroprevalence study was carried out in 50-60-ies of the past century in humans, agricultural animals and rodents. It was found that serotgroups Pomona, Icteroheamorrhagiae and Grippothypos prevailed in humans, Serogroup L. Pomona - dominated in animals, while in rodents the dominated serotgrouo was L. Icteroheamorrhagiae. Similar study, conducted by us in 1990-2005 showed that L.Pomona and L. Icteroheamorrhagiae prevailed in humans and rodents while L. Icteroheamorrhagiae was the only serogroup isolated in gray rats, trapped in Tbilisi. Infectivity of rodants composed 70%. Currently lesptosirosis is caused by a new serogroups of Leptospira - L. Sejroe, namely by serotypes Wolfii and Hardjo, L. Australis (serotype Bratislava) and L. Autumnalis along with the previously occurring serogroups L. Icteroheamorrhagiae, L. Bataviae, L. Hebdomadis. In our opinion the recent increase in leptospirosis in the country is attributable to the improvements in diagnostics of the infection as well as to the emergence of new serogroups of Leptospira, which in turn is a result of intensive migration processes in the country.


Diversity and Differentiation of Duckweed Species from Israel.
Duckweeds (Lemnaceae) are tiny plants that float on aquatic surfaces and are typically isolated from temperate and equatorial regions. Yet, duckweed diversity in Mediterranean and arid regions has been seldom explored. To address this gap in knowledge, we surveyed duckweed diversity in Israel, an ecological junction between Mediterranean and arid climates. We searched for duckweeds in the north and center of Israel on the surface of streams, ponds and waterholes. We collected and isolated 27 duckweeds and characterized their morphology, molecular barcodes (atpF-atpH and psbK-psbI) and biochemical features (protein content and fatty acids composition). Six species were identified-Lemna minor, L. gibba and Wolffia arrhiza dominated the duckweed populations, and together with past sightings, are suggested to be native to Israel. The fatty acid profiles and protein content further suggest that diverged functions have attributed to different haplotypes among the identified species. Spirodela polyrhiza, W. globosa and L. minuta were also identified but were rarer. S. polyrhiza was previously reported in our region, thus, its current low abundance should be revisited. However, L. minuta and W. globosa are native to America and Far East Asia, respectively, and are invasive in Europe. We hypothesize that they may be invasive species to our region as well, carried by migratory birds that disperse them through their migration routes. This study indicates that the duckweed population in Israel's aquatic environments consists of both native and transient species.


Interventional musculoskeletal procedures performed by using MR imaging guidance with a vertically open MR unit: assessment of techniques and applicability.
To evaluate the safety of and time required for a broad range of musculoskeletal interventional procedures performed by using magnetic resonance (MR) imaging guidance with a vertically open 0.5-T unit. Sixty-three MR imaging-guided procedures were performed. A vertically open MR unit equipped with in-room display monitors allowed interactive freehand MR guidance predominantly with fast spin-echo and gradient-echo sequences. Each procedure was classified in terms of the anatomic location, procedure type, and tissue type involved. The procedures were evaluated for success of needle placement, adequacy of tissue sampling, total procedural time, needle time, number of needle passes, and complications. Procedures consisted of tissue sampling with core-needle (n = 6) or fine-needle aspiration (n = 20) biopsy, corticosteroid or contrast agent injection (n = 19), joint cyst aspiration (n = 7), and drainage (n = 11). Successful needle placement was achieved in all 63 cases. Cytologic and histologic tissue samples were sufficient for pathologic diagnosis in 24 of 26 cases. In two cases, complications occurred: transient local bleeding and a brief vasovagal episode. The mean total procedural time was 64.8 minutes; the mean needle time, 26.2 minutes; and the mean number of needle passes per patient, 1.6. With use of a vertically open MR unit, MR-guided interventional procedures involving bone, soft tissue, intervertebral disks, and joints are safe and sufficiently rapid for use in clinical practice.


Nonlinear optical properties of Ni(Me(6)pzS(2))MX (M = Ni, Pd, Pt; X = Me(2)timdt, mnt).
In this work we investigate the second-order response of complexes of formula Ni(Me(6)pzS(2))MX (M = Ni, Pd, Pt; X = Me(2)timdt (monoanion of N,N'-disubstituted imidazolidine-2,4,5-trithione), mnt (maleonitriledithiolate)): by binding the porphyrazine to the metal-dithiolene, the electron asymmetry and pi-conjugation of the latter is increased, and its second-order response can result enhanced. By performing ab initio calculations of the ground-state and response properties of these compounds, we predict the molecular first hyperpolarizability, we elucidate its electronic origin, and we illustrate its dependence on the metal and the dithiolene ligand. Our study indicates that these complexes show a very high second-order response, comparable to that of organic "push-pull" materials, and that the appropriate metal-dithiolene combination can significantly enhance it.


A meta-analysis of plant facilitation in coastal dune systems: responses, regions, and research gaps.
Empirical studies in salt marshes, arid, and alpine systems support the hypothesis that facilitation between plants is an important ecological process in severe or 'stressful' environments. Coastal dunes are both abiotically stressful and frequently disturbed systems. Facilitation has been documented, but the evidence to date has not been synthesized. We did a systematic review with meta-analysis to highlight general research gaps in the study of plant interactions in coastal dunes and examine if regional and local factors influence the magnitude of facilitation in these systems. The 32 studies included in the systematic review were done in coastal dunes located in 13 countries around the world but the majority was in the temperate zone (63%). Most of the studies adopt only an observational approach to make inferences about facilitative interactions, whereas only 28% of the studies used both observational and experimental approaches. Among the factors we tested, only geographic region mediates the occurrence of facilitation more broadly in coastal dune systems. The presence of a neighbor positively influenced growth and survival in the tropics, whereas in temperate and subartic regions the effect was neutral for both response variables. We found no evidence that climatic and local factors, such as life-form and life stage of interacting plants, affect the magnitude of facilitation in coastal dunes. Overall, conclusions about plant facilitation in coastal dunes depend on the response variable measured and, more broadly, on the geographic region examined. However, the high variability and the limited number of studies, especially in tropical region, indicate we need to be cautious in the generalization of the conclusions. Anyway, coastal dunes provide an important means to explore topical issues in facilitation research including context dependency, local versus regional drivers of community structure, and the importance of gradients in shaping the outcome of net interactions.


Preoperative assessment of cutaneous melanoma thickness by multispectral dermoscopy.
Preoperative assessment of Breslow thickness by means of sonography and clinical and dermoscopic criteria in white light dermoscopy has been reported, but up until now, the use of multispectral dermoscopy has not been investigated. Aim of this research is to determine whether multispectral dermoscopy and more specifically pigment maps can be used as a predictive marker for Breslow thickness in melanoma. Pigment maps are generated in real time from multispectral dermoscopic images and help to visualize the presence of pigment in a lesion. Multispectral images of 110 melanomas were collected, using a digital handheld multispectral dermatoscope, and assessed independently by five observers for the presence or absence of deep pigment compared with the surrounding skin. According to histopathological examination, the mean Breslow thickness of all 110 melanomas was 1.04 mm (ranging from 0.1 to 14 mm). The group of melanomas where deep pigment was visualized on the multispectral image (n = 78) had a significantly higher Breslow thickness (1.19 mm) than the group where no deep pigment was observed (n = 32, mean Breslow 0.68 mm) (P = 0.025). This study is unique in preoperative assessment of tumour thickness by means of multispectral dermoscopy. Our data indicate that the presence of deep pigment as visualized in digital dermoscopic skin parameter maps identifies a group of thicker melanomas. Further prospective research is needed to validate these pigment maps, generated by multispectral dermoscopy as a measure to predict invasiveness in melanoma.


Retinol improves in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm during the first wave of spermatogenesis.
Testicular tissue freezing has been proposed for fertility preservation in pre-pubertal boys. Thawed frozen testicular tissue must undergo a maturation process to restore sperm production. The purpose of the current study was to evaluate the ability of retinol to improve the in vitro differentiation of pre-pubertal mouse spermatogonial stem cells into sperm. Testes from pre-pubertal mice, aged 2.5 and 6.5 days post-partum, were cultured on agarose gel at a gas-liquid interphase for 34, 38 and 60 days (D) and for 16, 30 and 36 D respectively. Assessment of basal medium (BM) supplemented with retinol (RE) alone, FSH/LH alone or a combination of both, was performed. Stereological analyses and tissue lesion scoring were performed at the culture time points indicated above. Sperm production was quantified at D30 and D34 after mechanical dissection of the testicular tissues. FSH/LH significantly increased the percentage of round spermatids at D30 and D38, when compared to BM alone. However, RE significantly increased the percentages of round but also elongated spermatids at D30 and D34. Moreover, RE significantly increased the number of spermatozoa per milligram of tissue at D30 and D34 when compared to BM. Therefore, RE improved the in vitro production of spermatids and spermatozoa from pre-pubertal SSCs during the first wave of spermatogenesis. The use of RE could be a useful tool for in vitro spermatogenesis from pre-pubertal human testicular tissue.


Measurement of Na+-K+ coupling ratio of Na+-K+-ATPase in rabbit proximal tubules.
A combination of 23Na nuclear magnetic resonance (NMR) spectroscopy and a K+-selective electrode was used to make simultaneous measurements of net Na+ and K+ fluxes across plasma membranes of rabbit renal proximal tubules after an abrupt stimulation of Na+-K+-ATPase. After a step in extracellular K+ concentration ([K+]o) from low to higher concentration (0.1-0.3 mM to 0.5-5.2 mM) at 25 degrees C, net extrusion of Na+ and uptake of K+ were observed. These fluxes were completely inhibited by ouabain (10(-3) M). Because initial rates of K+ uptake in presence or absence of Ba2+ (a known inhibitor of plasma membrane K+ conductance) were indistinguishable, net K+ flux was virtually unidirectional. Because suspension buffers contained neither glucose nor amino acids and the ratio of net Na+ and K+ fluxes (JNa and JK, respectively) was constant over a wide range of transmembrane Na+ gradients and absolute values of the JNa and JK, it is likely that changes in electrogenic or passive net fluxes across plasma membranes were insignificant in the first 30-45 s after the [K+]o step. Thus the ratio of these initial net Na+ and K+ fluxes corresponds closely to the Na+-K+ coupling ratio of the Na+-K+-ATPase. In 12 experiments, the measured Na+-K+-ATPase coupling ratio was 1.54 +/- 0.07 (SE). The coupling ratio was constant over a wide range of intracellular Na+ content, intracellular sodium concentration, [K+]o and transmembrane Na+ gradient. The coupling ratio also remained constant over an eightfold range of Na+-K+-ATPase rates.


Analysis of para-aortic lymphadenectomy up to the level of the renal vessels in apparent early-stage ovarian cancer.
The aim of this study was to evaluate the impact of para-aortic lymphadenectomy up to the renal vessels on the accurate staging in ovarian cancer patients presumed preoperatively to be confined to the ovary. We retrospectively analyzed data on 124 patients with primary epithelial ovarian cancer who were preoperatively thought to have tumor confined to the ovary and underwent primary staging surgery. The distribution of lymph node metastasis and various risk factors for nodal involvement were investigated. SURGICAL STAGING YIELDED: 87 (70.2%) patients had International Federation of Gynecology and Obstetrics (FIGO) stage I disease and 37 (29.8%) patients had stage II-III disease: 4 IIA, 6 IIB, 9 IIC, 1 IIIA, and 17 IIIC. Eighty-six patients had pelvic lymphadenectomy only and 69 had pelvic and para-aortic lymphadenectomy. Lymph node metastases were found in 17 (24.6%) of 69 patients; 5 (7.2%) patients had lymph node metastasis in the pelvic lymph nodes only, 8 (11.6%) in the para-aortic lymph nodes only, and 4 (5.8%) in both pelvic and para-aortic lymph nodes. Six (8.7%) patients had lymph node metastasis in the para-aortic lymph node above the level of the inferior mesenteric artery. On multivariate analysis, grade 3 tumor (p=0.01) and positive cytology (p=0.03) were independent predictors for lymph node metastasis. A substantial number of patients with apparently early ovarian cancer had upstaged disease. Of patients who underwent lymphadenectomy, some patients had lymph node metastasis above the level of the inferior mesenteric artery. Para-aortic lymphadenectomy up to the renal vessels may detect occult metastasis and be of help in tailoring appropriate adjuvant treatment as well as giving useful information about the prognosis.


Relationship between nursing students' preference for types of teaching materials and learning effects of self-learning tool.
The purpose of this study was to investigate the relationship between learning effects of the self-learning tool for nursing students and types of teaching materials. Ten nursing students were asked to perform transfer a patient from bed to wheelchair after watching the demo video and practicing 20 minutes. The students' performance was evaluated before and after practicing. The students were also asked to choose teaching materials that would be developed in the future. Out of nine teaching materials, the students chose seven of them. Correspondence analysis was conducted between the results of the evaluation of students' transfer technique and their preference of teaching materials. The results indicated that there was no relationship between the preference of teaching materials and the scores of transfer techniques. The authors concluded that the self-learning was not affected by the preference of teaching materials.


Prevotella jejuni sp. nov., isolated from the small intestine of a child with coeliac disease.
Five obligately anaerobic, Gram-stain-negative, saccharolytic and proteolytic, non-spore-forming bacilli (strains CD3 : 27, CD3 : 28(T), CD3 : 33, CD3 : 32 and CD3 : 34) are described. All five strains were isolated from the small intestine of a female child with coeliac disease. Cells of the five strains were short rods or coccoid cells with longer filamentous forms seen sporadically. The organisms produced acetic acid and succinic acid as major metabolic end products. Phylogenetic analysis based on comparative 16S rRNA gene sequence analysis revealed close relationships between CD3 : 27, CD3 : 28(T) and CD3 : 33, between CD3 : 32 and Prevotella histicola CCUG 55407(T), and between CD3 : 34 and Prevotella melaninogenica CCUG 4944B(T). Strains CD3 : 27, CD3 : 28(T) and CD3 : 33 were clearly different from all recognized species within the genus Prevotella and related most closely to but distinct from P. melaninogenica. Based on 16S rRNA, RNA polymerase β-subunit (rpoB) and 60 kDa chaperonin protein subunit (cpn60) gene sequencing, and phenotypic, chemical and biochemical properties, strains CD3 : 27, CD3 : 28(T) and CD3 : 33 are considered to represent a novel species within the genus Prevotella, for which the name Prevotella jejuni sp. nov. is proposed. Strain CD3 : 28(T) ( = CCUG 60371(T) = DSM 26989(T)) is the type strain of the proposed novel species. All five strains were able to form homologous aggregates, in which tube-like structures were connecting individual bacteria cells. The five strains were able to bind to human intestinal carcinoma cell lines at 37 °C.


New oligopeptide immunoglobulin G test for human parvovirus B19 antibodies.
A new, highly sensitive and specific enzyme immunoassay using oligopeptides as antigen (enzyme-linked immunosorbent assay [ELISA] B19-OP) for detecting parvovirus B19-specific immunoglobulin G (IgG) was established. As antigens, B19-specific oligopeptides of 24 and 30 kDa derived from a 196-kDa fusion protein of beta-galactosidase and viral capsid protein (VPI) of B19 after CNBr cleavage and separation by high-pressure liquid chromatography were used. Of 139 serum specimens tested in parallel for anti-B19 IgG by standard ELISA using B19 particles as antigen and by ELISA B19-OP, 73 (52.5%) were positive and 63 (45.3%) were negative in both tests, and 3 (2.2%) were negative by standard ELISA but positive by ELISA B19-OP and by immunoblot. By using ELISA B19-OP, it was possible to detect anti-B19 IgG in an asymptomatic blood donor 4 weeks after acute infection, and anti-B19 IgG titers of 10(-5) could be measured in convalescent-phase sera.


Design and baseline characteristics of a study of primary prevention of coronary events with pravastatin among Japanese with mildly elevated cholesterol levels.
Although cholesterol management reportedly reduces fatal and non-fatal coronary heart disease (CHD) events in subjects with or without evident atherosclerotic disease, it is still uncertain whether these benefits extend to Japanese. The study group comprised 8,009 subjects with mildly elevated total cholesterol who were randomized to treatment with 10-20 mg pravastatin plus diet (2,691 women, 1,267 men) or diet alone (2,758 women, 1,293 men). The groups were extremely well balanced with respect to baseline demographics and risk factors such as blood pressure and plasma lipids. Over a 5-year period of follow-up, the primary end-points will be a composite of fatal and non-fatal coronary events. Secondary end-points will include stroke and transient ischemic attack, all cardiovascular events and total mortality. The 2 groups will be followed up until the end of March 2004 and end-points will be analyzed by full analysis set.


DNA-Protein Cross-Link Formation in Nucleosome Core Particles Treated with Methyl Methanesulfonate.
N7-Methyl-2'-deoxyguanosine (MdG) is the major damage product in DNA produced by methylating agents, but it often thought to be nontoxic and nonmutagenic. MdG is chemically unstable. An abasic site (AP) is the major product produced from MdG under physiologically relevant conditions. AP formation is frequently considered to be responsible for the cytotoxic effects of MdG, but the reaction is suppressed in nucleosome core particles (NCPs). Recently, it was discovered that histone proteins form reversible DNA-protein cross-links (DPCs) with MdG in reconstituted NCPs, as well as in methylmethanesulfonate (MMS) treated cells. In this study, the formation and reactivity of MdG in MMS treated NCPs was examined at single nucleotide resolution. Sequences consisting of three or more consecutive dGs are more reactive with MMS. The efficiency and selectivity of MdG formation by MMS is largely unaffected within a NCP, although reactivity at several dGs is ∼1.5-2.5-fold higher in NCPs. DPC formation from MdG (DPCMdG) predominates over AP at all positions within the NCP. With few exceptions, DPCMdG yield is strongly dependent upon the accessibility of the major groove containing MdG to lysine-rich histone N-terminal tails. These data indicate that histone-MdG DPC formation will depend upon DNA sequence and translational position within an NCP.


Intentional replantation of periodontally hopeless teeth using a combination of enamel matrix derivative and demineralized freeze-dried bone allograft.
This study evaluated the clinical and radiographic results of the intentional replantation of periodontally hopeless teeth with combined enamel matrix derivative and demineralized freeze-dried bone allograft therapy. Eleven patients (five female, six male; age range, 13 to 53 years) with 12 periodontally hopeless teeth resulting from extensive alveolar bone loss and vertical defects extending to the apexes were studied. At the 12-month clinical and radiologic follow-up, significant improvement was observed for all clinical and radiographic parameters except gingival recession (P < .05). These preliminary findings show that intentional replantation combined with regenerative techniques is a successful alternative to tooth extraction.


Inkjet-Printed Molybdenum Disulfide and Nitrogen-Doped Graphene Active Layer High On/Off Ratio Transistors.
Fully inkjet-printed device fabrication is a crucial goal to enable large-area printed electronics. The limited number of two-dimensional (2D) material inks, the bottom-gated structures, and the low current on/off ratio of thin-film transistors (TFTs) has impeded the practical applications of the printed 2D material TFTs. In the search for TFTs with high current ratios, we introduce a stable and efficient method of nitrogen-doped graphene (NDG) ink preparation for inkjet printing by liquid-phase exfoliation. The NDG thin film is print-stacked with molybdenum disulfide (MoS2) by multiple printing passes to construct a MoS2-NDG stack. We demonstrate top-gated fully inkjet-printed MoS2-NDG transistors with silver drain, source, and gate electrodes, and a barium titanate (BaTiO3) dielectric. A 100% inkjet-printed MoS2-NDG vertical 2D active heterostructure layer transistor with a current on/off ratio of 1200 is exhibited. The results may lead towards the development of all-printed 2D material-based transistor switches.


Impact of pH on the activity of novel cephalosporin cefiderocol in human urine.
Antimicrobial activity of antibiotics can be impacted by pH, enhancing or reducing their bactericidal properties. Cefiderocol, a novel cephalosporin antibiotic that is among others indicated for the treatment of complicated urinary tract infections (cUTIs), lacks data on activity in urine. Pooled human urine (iron levels ∼0.05 mg/L/24 h), CAMHB and iron-depleted CAMHB (ID-CAMHB) at pH 5, 7 and 8 served as media. MIC testing was done according to EUCAST with the broth microdilution method for 17 clinical isolates of Escherichia coli and ATCC 25922 (including isolates with ESBL activity), 17 clinical isolates of Klebsiella pneumoniae and ATCC 700603 (also with ESBL), and 6 clinical isolates of Pseudomonas aeruginosa and ATCC 27853. Time-kill curves (TKCs) were performed for selected strains at pH 5, 7 and 8 in urine. MIC values in urine, CAMHB and ID-CAMHB exhibited isolate-specific variations when assessed under identical pH conditions, ranging from a 1-fold dilution to changes of up to 4-fold dilutions in either direction. Median MICs of cefiderocol were up to 50-fold higher in pH 5 than in pH 7 for P. aeruginosa isolates and 32-fold higher in E. coli and K. pneumoniae isolates. TKCs with 650 and 1300 mg/L cefiderocol in urine showed that ATCC strains were efficiently eradicated despite the pH set. Acidic pH had a significant negative impact on cefiderocol activity. Yet, after a recommended IV administration of 2 g cefiderocol every 8 h, a concentration of approximately 1300 mg/L can be achieved in urine, suggesting that efficient killing of all tested pathogens could have been possible even under acidic conditions in vivo.


Cysteine-conjugated metabolites of ginger components, shogaols, induce apoptosis through oxidative stress-mediated p53 pathway in human colon cancer cells.
Shogaols, the major constituents of thermally processed ginger, have been proven to be highly effective anticancer agents. Our group has identified cysteine-conjugated shogaols (M2, M2', and M2″) as the major metabolites of [6]-, [8]-, and [10]-shogaol in human and found that M2 is a carrier of its parent molecule [6]-shogaol in cancer cells and in mice, while being less toxic to normal colon fibroblast cells. The objectives of this study are to determine whether M2' and M2″ behave in a similar manner to M2, in both metabolism and efficacy as anticancer agents, and to further explore the biological pro-apoptotic mechanisms of the cysteine-conjugated shogaols against human colon cancer cells HCT-116 and HT-29. Our results show that [8]- and [10]-shogaol have similar metabolic profiles to [6]-shogaol and exhibit similar toxicity toward human colon cancer cells. M2' and M2″ both show low toxicity against normal colon cells but retain potency against colon cancer cells, suggesting that they have similar activity to M2. We further demonstrate that the cysteine-conjugated shogaols can cause cancer cell death through the activation of the mitochondrial apoptotic pathway. Our results show that oxidative stress activates a p53 pathway that ultimately leads to p53 up-regulated modulator of apoptosis (PUMA) induction and down-regulation of B-cell lymphoma 2 (Bcl-2), followed by cytochrome c release, perturbation of inhibitory interactions of X-linked inhibitor of apoptosis protein (XIAP) with caspases, and finally caspase 9 and 3 activation and cleavage. A brief screen of the markers attenuated by the proapoptotic activity of M2 revealed similar results for [8]- and [10]-shogaol and their respective cysteine-conjugated metabolites M2' and M2″. This study highlights the cysteine-conjugated metabolites of shogaols as novel dietary colon cancer preventive agents.


Dietary peptide-specific antibodies against interleukin-4 differentially alter systemic immune cell responses during Eimeria challenge with minimal impacts on the cecal microbiota.
Eimeria spp. induce host interleukin (IL)-4 production, a potent immune regulator, during coccidiosis to evade immune responses. Dietary anti-IL-4 may preserve bird performance during challenge; however, specific mechanisms have not been investigated. Study objectives were to develop peptide-specific anti-IL-4 antibodies and evaluate immune cell profiles and the cecal microbiota during Eimeria challenge. Four candidate IL-4 peptides were selected based on antigenicity and location. Hens were injected with conjugated peptide or carrier-only control (3/injection), eggs were collected post-vaccination and yolks were pooled by peptide before freeze-drying. On d 0, 300 Ross 708 broilers were placed in floor pens (10/pen) and assigned to 5 diets consisting of basal diet + 2% egg yolk powder containing antibodies against 1 of 4 target peptides or carrier-only control for 14-d starter and grower periods (28 d total). Baseline blood and cecal contents were collected on d 14 (6 birds/diet) before half the remainder were inoculated with 10X Coccivac-B52 (Merck Animal Health, Kenilworth, NJ). Body weight (BW) and feed intake (FI) were recorded weekly and blood and cecal samples were collected at 3, 7, and 14 d post-inoculation (pi; 3/treatment). Immune cell profiles in peripheral blood mononuclear cells (PBMC) were evaluated flow cytometrically and cecal microbial communities determined by 16S/18S rRNA gene amplicon sequencing. Data were log-transformed when necessary and analyzed with diet, Eimeria, and timepoint fixed effects plus associated interactions (SAS 9.4; P ≤ 0.05). Anti-IL-4 did not alter baseline performance but generally increased PBMC Bu-1+ B cells 38.0 to 55.4% (P < 0.0001). Eimeria challenge reduced FI and BWG 16.1 and 30.3%, respectively, regardless of diet (P < 0.0001) with only birds fed peptide 4 antibodies not recovering feed conversion by d 28. Minimal diet-associated cecal microbiota changes were observed, indicating that anti-IL-4 effects were likely host-specific. Eimeria-challenged birds fed peptide 3 antibodies displayed minimal immune cell fluctuations compared to unchallenged counterparts, suggesting these antibodies potentially modulated intestinal immune responses to minimize systemic requirements, making them good candidates for further research.


Mycobacterium tuberculosis gene expression during adaptation to stationary phase and low-oxygen dormancy.
The innate mechanisms used by Mycobacterium tuberculosis to persist during periods of non-proliferation are central to understanding the physiology of the bacilli during latent disease. We have used whole genome expression profiling to expose adaptive mechanisms initiated by M. tuberculosis in two common models of M. tuberculosis non-proliferation. The first of these models was a standard growth curve in which gene expression changes were followed from exponential growth through the transition to stationary phase. In the second model, we followed the adaptive process of M. tuberculosis during transition from aerobic growth to a state of anaerobic non-replicating persistence. The most striking finding from these experiments was the strong induction of the entire DosR "dormancy" regulon over approximately 20 days during the long transition to an anaerobic state. This is contrasted by the muted overall response to aerated stationary phase with only a partial dormancy regulon response. From the results presented here we conclude that the respiration-limited environment of the oxygen-depleted NRP model recreates at least one fundamental factor for which the genome of M. tuberculosis encodes a decisive adaptive program.


Total knee replacement for patients with patellofemoral arthritis.
Fifty-three patients with incapacitating patellofemoral arthritis and minimal tibiofemoral changes underwent a total knee replacement. Their results were compared with the results of a concomitant series of patients with tricompartmental osteoarthritis. Preoperatively group of patients with patellofemoral arthritis had more individuals with a lateral patellar tilt and inability to climb stairs in a bipedal manner than in the group with tricompartmental arthritis. Furthermore, fewer patients in the group of patients with patellofemoral arthritis could rise independently from a chair than could those patients with tricompartmental arthritis. The mean followup period was 7.4 years. The rate of lateral parapatellar releases was three times greater in the patients with patellofemoral arthritis. The group of patients with patellofemoral arthritis had higher knee scores and a greater ability to climb stairs in a bipedal manner as compared with the patients with tricompartmental arthritis. Residual anterior knee pain was present in approximately 7% of patients in both groups. Total knee replacement resulted in better outcomes for older patients with patellofemoral arthritis as compared with patellofemoral replacement, debridement procedures, and patellectomy.


IL-6 blockade in the management of non-infectious uveitis.
Several pathogenetic studies have paved the way for a newer more rational therapeutic approach to non-infectious uveitis, and treatment of different forms of immune-driven uveitis has drastically evolved in recent years after the advent of biotechnological drugs. Tumor necrosis factor-α targeted therapies, the first-line recommended biologics in uveitis, have certainly led to remarkable results in patients with non-infectious uveitis. Nevertheless, the decision-making process turns out to be extremely difficult in anti-tumor necrosis factor or multidrug-resistant cases. Interleukin (IL)-6 holds a critical role in the pathogenic pathways of uveitis, due to its extended and protean range of effects. On this background, manipulation of IL-6 inflammatory cascade has unraveled encouraging outcomes. For instance, rising evidence has been achieved regarding the successful use of tocilizumab, the humanized monoclonal antibody targeted against the IL-6 receptor, in treating uveitis related to juvenile idiopathic arthritis or Behçet's disease. Similar findings have also been reported for uveitis associated with systemic disorders, such as rheumatoid arthritis or multicentric Castleman disease, but also for idiopathic uveitis, the rare birdshot chorioretinopathy, and even in cases complicated by macular edema. This work provides a digest of all current experiences and evidences concerning IL-6 blockade, as suggested by the medical literature, proving its potential role in the management of non-infectious uveitis.


Intersession test-retest variability of 10-2 MAIA microperimetry in fixation-threatening glaucoma.
To determine the intersession test-retest variability (TRV) of CenterVue Macular Integrity Assessment (MAIA) microperimeter in glaucoma patients with fixation-threatening field defects. This is a prospective case-control study of 27 participants consisting of 13 patients with stable primary open-angle glaucoma and 14 control subjects including 5 healthy individuals and 9 retinal patients (5 with non-neovascular age-related macular degeneration and 4 with inherited retinal disease). Each participant underwent three microperimetry tests in one eye at 1-month intervals. Each test used an identical test strategy of 10-2 Cartesian grid and 4-2 staircase algorithm. We investigated TRV by calculating the coefficient of repeatability (CR) for mean sensitivity (MS) and point-wise sensitivity (PWS) for glaucomatous subjects and retinal and normal subjects. 95% confidence intervals (CIs) for CRs were calculated. There was no significant change in MS, and the median durations of microperimetry sessions were 9'26″, 8'52″, and 8'46″ across the three study visits. The intersession CRs for MS were 1.1, 2.5, and 1.8 dB, and the average CRs for PWS were 3.5, 7.4, and 8.6 dB for healthy controls and retinal and glaucoma patients, respectively. For test loci with 25-34 dB at baseline, CRs for PWS were 8.2 (95% CI: 7.5-8.9) and 4.3 (95% CI: 4.0-4.6) dB for glaucoma and control subjects, respectively. We found differences in TRV of test loci depending on the baseline sensitivity value. Glaucoma patients had significantly worse TRV for loci that had sensitivity values within the normal range at baseline. The estimated CR has implications for sample size calculation in future glaucoma treatment trials using microperimetry as a clinical endpoint.


An LCMS-based untargeted metabolomics protocol for cochlear perilymph: highlighting metabolic effects of hydrogen gas on the inner ear of noise exposed Guinea pigs.
Noise-induced hearing loss (NIHL) is an increasing problem in society and accounts for a third of all cases of acquired hearing loss. NIHL is caused by formation of reactive oxygen species (ROS) in the cochlea causing oxidative stress. Hydrogen gas (H2) can alleviate the damage caused by oxidative stress and can be easily administered through inhalation. To present a protocol for untargeted metabolomics of guinea pig perilymph and investigate the effect of H2 administration on the perilymph metabolome of noise exposed guinea pigs. The left ear of guinea pigs were exposed to hazardous impulse noise only (Noise, n = 10), noise and H2 (Noise + H2, n = 10), only H2 (H2, n = 4), or untreated (Control, n = 2). Scala tympani perilymph was sampled from the cochlea of both ears. The polar component of the perilymph metabolome was analyzed using a HILIC-UHPLC-Q-TOF-MS-based untargeted metabolomics protocol. Multivariate data analysis (MVDA) was performed separately for the exposed- and unexposed ear. MVDA allowed separation of groups Noise and Noise + H2 in both the exposed and unexposed ear and yielded 15 metabolites with differentiating relative abundances. Seven were found in both exposed and unexposed ear data and included two osmoprotectants. Eight metabolites were unique to the unexposed ear and included a number of short-chain acylcarnitines. A HILIC-UHPLC-Q-TOF-MS-based protocol for untargeted metabolomics of perilymph is presented and shown to be fit-for-purpose. We found a clear difference in the perilymph metabolome of noise exposed guinea pigs with and without H2 treatment.


Activation of natural killer T cells by alpha-galactosylceramide rapidly induces the full maturation of dendritic cells in vivo and thereby acts as an adjuvant for combined CD4 and CD8 T cell immunity to a coadministered protein.
The maturation of dendritic cells (DCs) allows these antigen-presenting cells to initiate immunity. We pursued this concept in situ by studying the adjuvant action of alpha-galactosylceramide (alphaGalCer) in mice. A single i.v. injection of glycolipid induced the full maturation of splenic DCs, beginning within 4 h. Maturation was manifest by marked increases in costimulator and major histocompatibility complex class II expression, interferon (IFN)-gamma production, and stimulation of the mixed leukocyte reaction. These changes were not induced directly by alphaGalCer but required natural killer T (NKT) cells acting independently of the MyD88 adaptor protein. To establish that DC maturation was responsible for the adjuvant role of alphaGalCer, mice were given alphaGalCer together with soluble or cell-associated ovalbumin antigen. Th1 type CD4+ and CD8+ T cell responses developed, and the mice became resistant to challenge with ovalbumin-expressing tumor. DCs from mice given ovalbumin plus adjuvant, but not the non-DCs, stimulated ovalbumin-specific proliferative responses and importantly, induced antigen-specific, IFN-gamma producing, CD4+ and CD8+ T cells upon transfer into naive animals. In the latter instance, immune priming did not require further exposure to ovalbumin, alphaGalCer, NKT, or NK cells. Therefore a single dose of alphaGalCer i.v. rapidly stimulates the full maturation of DCs in situ, and this accounts for the induction of combined Th1 CD4+ and CD8+ T cell immunity to a coadministered protein.


Insulin and oral agents for managing cystic fibrosis-related diabetes.
Insulin therapy is recommended by the Cystic Fibrosis Foundation when cystic fibrosis-related diabetes has been diagnosed. Diagnosis is based on an elevated fasting blood glucose level greater than 6.94 mmol/liter (125 mg/deciliter) or symptomatic diabetes for random glucose levels greater than 11.11 mmol/liter (200 mg/deciliter). To examine the evidence that, when treated with agents for managing diabetes, people with cystic fibrosis improve their sugar metabolic control resulting in beneficial impact on lung function and the ability to maintain optimal weight. We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register comprising references identified from comprehensive electronic database searches and handsearches of relevant journals and abstract books of conference proceedings. We also handsearched abstracts from pulmonary and North American Cystic Fibrosis Conference symposia. Date of the most recent search of the Group's Trials Register: December 2004. Randomized controlled trials comparing all methods of diabetes therapy for one month or longer in people with cystic fibrosis. No studies were found which were eligible for inclusion in this review. Six references to four studies were identified by the searches, but none were eligible for inclusion in the review as they were not randomized controlled trials. While some cystic fibrosis centers use oral medications to help control diabetes, a condition which complicates the course of cystic fibrosis, insulin therapy is the recommended and most widely used treatment method. Lung function has been reported to improve with the use of insulin, but this has not been correlated to the degree in which sugar metabolism has been affected. While the Cystic Fibrosis Foundation recommends insulin therapy be used in managing diabetes, this systematic review identifies the need for a multicentre randomized controlled trial assessing both the efficacy of insulin or other insulin-releasing or insulin-sensitizing medications and their possible adverse effects in managing cystic fibrosis-related diabetes.


Regulation of estrogen and progestin receptor concentrations in an experimental rat prostatic carcinoma by estrogen, antiestrogen, and progesterone.
In order to assess prostatic tissue as a target for receptor-mediated estrogen action, we have examined the regulation of estrogen (ER) and progestin receptors (PgR) by estrogen, antiestrogen, and progesterone in cytosolic and nuclear fractions of the R3327H (Dunning) prostatic adenocarcinoma of the rat. Twenty micrograms diethylstilbestrol (DES) with or without 800 micrograms tamoxifen (Tam) were injected s.c. in oil 5 times weekly for 2 weeks. Controls were given oil only. Estrogen receptor assays were carried out using [3H]estradiol and a hydroxylapatite exchange method. Progestin receptors were assayed using [3H]R5020 and dextran-coated charcoal to separate free and bound steroid. All binding data were evaluated by using Scatchard analysis. Treatment with DES depleted cytosolic ER, promoted association of ER with the nuclear fraction, and concomitantly increased PgR concentrations in amounts proportional to nuclear ER. Treatment with Tam alone resulted in higher nuclear ER concentrations than treatment with DES, but induced only one-fifth the amount of PgR. Treatment with DES plus Tam resulted in similar nuclear ER concentrations as with Tam alone, but PgR concentrations were intermediate between those observed with DES alone and Tam alone. Thus Tam exhibited both estrogenic and antiestrogenic properties. In this experiment, the same cytosolic and nuclear extracts were also assayed for ER by using monoclonal antibodies to the receptor in an enzyme immunoassay. No significant differences were observed between the results obtained by the radioligand and enzyme immunoassay methods in the cytosol and nuclear fractions from the control and DES-treated tumors. However in both Tam-treated groups, the ER values obtained by the enzyme immunoassay method were significantly higher than those obtained by the radioligand method in both cytosolic and nuclear fractions. This confirms the observations made by others in female target organs, that monoclonal antibody to ER reacts differently with the Tam-bound ER complex than with the estradiol-bound ER complex. In a separate experiment, administration of progesterone with DES decreased the concentration of nuclear ER to less than one-half that observed after administration of DES alone, with proportional decreases in both cytosolic and nuclear PgR. All these observations indicate that the control of ER and PgR concentrations in this prostatic tumor is identical to that observed in female rat target organs. Use of an immunohistochemical method for the detection of ER in frozen sections indicated that the receptor was localized in the glandular epithelium in both control and DES-treated tumors.


The Rhomboid Protease GlpG Promotes the Persistence of Extraintestinal Pathogenic Escherichia coli within the Gut.
Extraintestinal pathogenic Escherichia coli (ExPEC) strains are typically benign within the mammalian gut but can disperse to extraintestinal sites to cause diseases like urinary tract infections and sepsis. As occupation of the intestinal tract is often a prerequisite for ExPEC-mediated pathogenesis, we set out to understand how ExPEC colonizes this niche. A screen using transposon sequencing (Tn-seq) was performed to search for genes within ExPEC isolate F11 that are important for growth in intestinal mucus, which is thought to be a major source of nutrients for E. coli in the gut. Multiple genes that contribute to ExPEC fitness in mucus broth were identified, with genes that are directly or indirectly associated with fatty acid beta-oxidation pathways being especially important. One of the identified mucus-specific fitness genes encodes the rhomboid protease GlpG. In vitro, we found that the disruption of glpG had polar effects on the downstream gene glpR, which encodes a transcriptional repressor of factors that catalyze glycerol degradation. Mutation of either glpG or glpR impaired ExPEC growth in mucus and on plates containing the long-chain fatty acid oleate as the sole carbon source. In contrast, in a mouse gut colonization model in which the natural microbiota is unperturbed, the disruption of glpG but not glpR significantly reduced ExPEC survival. This work reveals a novel biological role for a rhomboid protease and highlights new avenues for defining mechanisms by which ExPEC strains colonize the mammalian gastrointestinal tract.


Proximal and total humerus reconstruction with the use of an aortograft mesh.
The shoulder is commonly affected by primary and metastatic tumors. Current surgical techniques for complex shoulder reconstruction frequently result in functional deficits and instability. A synthetic mesh used in vascular surgery has the biological properties to provide mechanical constraint and improve stability after tumor related shoulder reconstruction. We describe (1) surgical technique using a synthetic mesh during humerus reconstructions; (2) functional level defined as shoulder ROM of patients undergoing the procedure; (3) incidence of postoperative dislocation and shoulder instability; and (4) complications associated with the use of the device. We retrospectively reviewed 16 patients with proximal humerus replacements reconstructed with a synthetic mesh from February 2006 to July 2008. Patients were followed clinically and radiographically for a minimum of 13 months (mean, 26 months; range, 13-43 months). There were no shoulder dislocations at the latest followup. The mean shoulder flexion was 43° (range, 15°-170°) and mean shoulder abduction of 38 (range, 15°-110°). The mean operative time was 121 minutes (range, 80-170 minutes) and the mean blood loss was 220 mL (range, 50-750 mL). One patient had a superficial wound infection and none a deep infection requiring removal of the graft or prosthesis. The data suggest the use of a synthetic vascular mesh for proximal humerus reconstruction may reduce dislocations and facilitate soft tissue attachment and reconstruction after tumor resection.


Weekly and consecutive day neonatal intubation training: comparable on a pediatrics clerkship.
To determine whether medical student intubation proficiency with a neonatal mannequin differs according to weekly or consecutive day practice sessions during a six-week pediatric clerkship. From July 2010 through June 2011, the authors prospectively randomized 110 third-year medical students into three neonatal intubation practice groups: standard (control; no practice sessions), weekly (practice once/week for four consecutive weeks), or consecutive day (practice once/day for four consecutive days). At baseline, students performed intubation during individual sessions using a neonatal mannequin (SimNewB). Two reviewers, blinded to practice group, viewed videotapes of intubations and independently scored students on equipment selection, procedural skill steps, length of intubation attempts (in seconds), and the number of attempts (up to three) needed for a successful intubation. Videotaped individual final assessment intubation sessions during week six were evaluated in the same manner. Students in the weekly and consecutive day practice groups performed better at the final assessment on all variables than students in the standard group (P < .001), but over six weeks, the authors detected no differences between the two distributed practice formats for any outcomes of interest. Practice improved all aspects of neonatal intubation performance, including choosing the correct equipment, properly performing the skill steps, length of time to successful intubation, and success rate, for novice health care providers in a simulation setting. Over six weeks, neither practice format proved superior, but it remains unclear whether one format is superior for learning and skill retention over the long term or in actual practice.


Dopaminergic mediation of long-term behavioral effects of in utero drug exposure.
The influence of external agents on the developing brain has led to the hypothesis that, during critical periods of rapid development, such agents may have long-lasting effects on brain function and behavior. The dopaminergic systems are particularly suitable for examining the hypothesis, since the neural tracts using dopamine have been widely studied; their influence on behavior is, to some extent, understood; the tracts begin development in mice and rats late in gestation; and there are a number of drugs known to have particular effects on these systems. Literature is reviewed that supports the hypothesis that drugs affecting dopamine metabolism, when administered during the early development of the dopaminergic tracts, produce persistent changes both in the metabolism of dopamine and in behavior mediated by neural systems which utilize dopamine as a transmitter.


Evaluation of the efficacy and safety of three dosing regimens of agalsidase alfa enzyme replacement therapy in adults with Fabry disease.
Efficacy and safety of agalsidase alfa at 0.2 mg/kg weekly were compared with 0.2 mg/kg every other week (EOW). Exploratory analyses were performed for 0.4 mg/kg weekly. This was a 53-week, Phase III/IV, multicenter, open-label study (NCT01124643) in treatment-naïve adults (≥18 years) with Fabry disease. Inclusion criteria were left ventricular hypertrophy at baseline, defined as left ventricular mass indexed to height >50 g/m(2.7) for males and >47 g/m(2.7) for females. Primary endpoint was reduction of left ventricular mass indexed to height as assessed by echocardiography. Secondary endpoints included cardiac (peak oxygen consumption, 6-minute walk test, Minnesota Living with Heart Failure Questionnaire, New York Heart Association classification), renal (Modification of Diet in Renal Disease, estimated glomerular filtration rate), and biomarker (plasma globotriaosylceramide) assessments. Safety endpoints were adverse events and anti-agalsidase alfa antibodies. Twenty patients were randomized to 0.2 mg/kg EOW (mean age, 50.3 years; 70% male), 19 to 0.2 mg/kg weekly (51.8 years; 53% male), and 5 to 0.4 mg/kg weekly (49.4 years; 40% male). The mean change in left ventricular mass indexed to height by Week 53 in the 0.2-mg/kg EOW and weekly groups was 3.2 g/m(2.7) and 0.5 g/m(2.7), with no significant difference between groups. No clinically meaningful changes by Week 53 were found within or between the 0.2-mg/kg groups for peak oxygen consumption, 6-minute walk test, or Minnesota Living with Heart Failure Questionnaire. Two patients in each group improved by ≥1 New York Heart Association classification. No significant differences were found between 0.2 mg/kg EOW and weekly for mean change in estimated glomerular filtration rate (-1.21 mL/min/1.73 m(2) vs -3.32 mL/min/1.73 m(2)) or plasma globotriaosylceramide (-1.05 nmol/mL vs -2.13 nmol/mL), respectively. Infusion-related adverse events were experienced by 25% and 21% in the 0.2-mg/kg EOW and weekly groups. Tachycardia, fatigue, and hypotension were experienced by two or more patients overall. Anti-agalsidase alfa antibodies were detected in 11.4% of patients and neutralizing antibodies in 6.8%. Infusion-related reactions did not appear to be correlated with antibody status. No efficacy or safety differences were found when the approved EOW dosage of agalsidase alfa was increased to weekly administration. Exploratory analyses for 0.4 mg/kg weekly showed similar results.


Clinical features and risk factors of pulmonary oedema after enterovirus-71-related hand, foot, and mouth disease.
In Taiwan, from April to July, 1998, an epidemic of hand, foot, and mouth disease associated with enterovirus 71 (EV71) occurred with fatal complications. We did a clinical study of EV71-related diseases in Taiwan. We studied 154 children with virus-culture confirmed EV71 infection. Children were divided into three groups: 11 patients with pulmonary oedema; 38 patients with central nervous system (CNS) involvement and no pulmonary oedema; and 105 children without complications. We compared the clinical features, laboratory findings, risk factors, and outcome among these three groups. Nine children with pulmonary oedema had hand, foot, and mouth disease, one had herpangina, and one had febrile illness with eight children with limb weakness and one with limb hypesthesia. All children had had sudden onset of tachycardia, tachypnoea, and cyanosis 1-3 days after onset of the disease. Nine of 11 children died within 12 h of intubation; one child was braindead within 15 h and died 17 days after intubation; one child was in deep coma and died 3 months later. In children with CNS complication and no pulmonary oedema, one child died of pneumonia after 4 months of ventilator support and four children had sequelae. All 105 children without complications recovered. There was a significant association between CNS involvement and pulmonary oedema (odds ratio 12.4 [95% CI 2.6-60.1], p=0.001). Risk factors for pulmonary oedema after CNS involvement were hyperglycaemia, leucocytosis, and limb weakness. Hyperglycaemia was the most significant prognostic factor for pulmonary oedema (odds ratio 21.5 [3-159], p=0.003). EV71 can cause hand, foot, and mouth disease, CNS involvement with severe sequelae, and fatal pulmonary oedema. Hyperglycaemia is the most important prognostic factor.


The relationship between mean glandular dose and compressed breast thickness specified for Jordan.
The mean glandular dose (MGD) is a measurement used in mammography to assess the amount of radiation absorbed. By considering specific exposure radiation dose criteria, MGD ensures minimal radiation while maintaining image quality for detecting abnormalities. The relationship between MGD and compressed breast thickness (CBT) is commonly utilized in mammographic dose surveys. This study aims to estimate the MGD-CBT relationship based on patient age in Jordan through retrospective analysis. The analysis involved 3465 screening mammography images of women aged 40-80, divided into three age groups: 40-49, 50-64 and 65-80 years. Each group had a specific CBT range (16.5-156 mm). The results indicate that MGD ranges from 1.6 to 1.7 mGy across all three age groups, independent of CBT. Thus, a significant and positive correlation exists between MGD and CBT in all age groups.


Canagliflozin-current status in the treatment of type 2 diabetes mellitus with focus on clinical trial data.
Canagliflozin (CFZ) is a member of new class of glucose lowering agents, sodium-glucose co-transporter (SGLT) inhibitors, which got approval by food and drug administration. It has insulin independent action by blocking the transporter protein SGLT2 in the kidneys, resulting in urinary glucose excretion and reduction in blood glucose levels. In clinical trials, CFZ significantly decreased HbA1c level when administered either as monotherapy or as combined therapy with other anti-diabetic drugs. Intriguingly, it showed additional benefits like weight reduction and lowering of blood pressure. The commonly observed side effects were urinary and genital infections. It has exhibited favorable pharmacokinetic and pharmacodynamic profiles even in patients with renal and hepatic damage. Hence, this review purports to outline CFZ as a newer beneficial drug for type 2 diabetes mellitus.


Recurrent acute pancreatitis.
Recurrent acute pancreatitis (RAP) is commonly encountered, but less commonly understood clinical entity, especially idiopathic RAP, with propensity to lead to repeated attacks and may be chronic pancreatitis if attacks continue to recur. A great number of studies have been published on acute pancreatitis, but few have focused on RAP. Analysing the results of clinical studies focusing specifically on RAP is problematic in view due to lack of standard definitions, randomised clinical trials, standard evaluation protocol used and less post intervention follow-up duration. With the availability of newer investigation modalities less number of etiologies will remains undiagnosed. This review particularly is focused on the present knowledge in understanding of RAP.


A computational continuum model of poroelastic beds.
Despite the ubiquity of fluid flows interacting with porous and elastic materials, we lack a validated non-empirical macroscale method for characterizing the flow over and through a poroelastic medium. We propose a computational tool to describe such configurations by deriving and validating a continuum model for the poroelastic bed and its interface with the above free fluid. We show that, using stress continuity condition and slip velocity condition at the interface, the effective model captures the effects of small changes in the microstructure anisotropy correctly and predicts the overall behaviour in a physically consistent and controllable manner. Moreover, we show that the performance of the effective model is accurate by validating with fully microscopic resolved simulations. The proposed computational tool can be used in investigations in a wide range of fields, including mechanical engineering, bio-engineering and geophysics.


Longitudinal Trends in Body Mass Index Before and During the COVID-19 Pandemic Among Persons Aged 2-19 Years - United States, 2018-2020.
Obesity is a serious health concern in the United States, affecting more than one in six children (1) and putting their long-term health and quality of life at risk.* During the COVID-19 pandemic, children and adolescents spent more time than usual away from structured school settings, and families who were already disproportionally affected by obesity risk factors might have had additional disruptions in income, food, and other social determinants of health.† As a result, children and adolescents might have experienced circumstances that accelerated weight gain, including increased stress, irregular mealtimes, less access to nutritious foods, increased screen time, and fewer opportunities for physical activity (e.g., no recreational sports) (2,3). CDC used data from IQVIA's Ambulatory Electronic Medical Records database to compare longitudinal trends in body mass index (BMI, kg/m2) among a cohort of 432,302 persons aged 2-19 years before and during the COVID-19 pandemic (January 1, 2018-February 29, 2020 and March 1, 2020-November 30, 2020, respectively). Between the prepandemic and pandemic periods, the rate of BMI increase approximately doubled, from 0.052 (95% confidence interval [CI] = 0.051-0.052 to 0.100 (95% CI = 0.098-0.101) kg/m2/month (ratio = 1.93 [95% CI = 1.90-1.96]). Persons aged 2-19 years with overweight or obesity during the prepandemic period experienced significantly higher rates of BMI increase during the pandemic period than did those with healthy weight. These findings underscore the importance of efforts to prevent excess weight gain during and following the COVID-19 pandemic, as well as during future public health emergencies, including increased access to efforts that promote healthy behaviors. These efforts could include screening by health care providers for BMI, food security, and social determinants of health, increased access to evidence-based pediatric weight management programs and food assistance resources, and state, community, and school resources to facilitate healthy eating, physical activity, and chronic disease prevention.


Influence of ascorbic acid on the thermic effect of feeding in overweight and obese adult humans.
The thermic effect of feeding (TEF: increase in energy expenditure following acute energy intake) is an important physiological determinant of total daily energy expenditure and thus energy balance. Approximately 40% of TEF is believed to be mediated by sympathoadrenal activation and consequent beta-adrenergic receptor stimulation of metabolism. In sedentary adults, acute administration of ascorbic acid, a potent antioxidant, augments the thermogenic response to beta-adrenergic stimulation. We hypothesized that acute ascorbic acid administration augments TEF in sedentary overweight and obese adults. Energy expenditure was determined (ventilated hood technique) before and 4 h after consumption of a liquid-mixed meal (caloric equivalent 40% of resting energy expenditure (REE)) in 11 sedentary, overweight/obese adults (5 men, 6 women; age: 24 +/- 2 years; BMI: 28.5 +/- 1.0 kg/m(2) (mean +/- s.e.)) on two separate, randomly ordered occasions: during continuous intravenous administration of saline (placebo control) and/or ascorbic acid (0.05 g/kg fat-free mass). Acute ascorbic acid administration prevented the increase in plasma concentration of oxidized low-density lipoprotein in the postprandial state (P = 0.04), but did not influence REE (1,668 +/- 107 kcal/day vs.1,684 +/- 84 kcal/day; P = 0.91) or the area under the TEF response curve (33.4 +/- 2.4 kcal vs. 30.5 +/- 3.6 kcal; P = 0.52) (control vs. ascorbic acid, respectively). Furthermore, acute ascorbic acid administration had no effect on respiratory exchange ratio, heart rate, or arterial blood pressure in the pre- and postabsorptive states (all P > 0.64). These data imply that the attenuated TEF commonly observed with sedentary lifestyle and obesity is not modulated by ascorbic acid-sensitive oxidative stress.