Isolation and immunochemical characterization of rabbit 7S anti-IgG with restricted heterogeneity.
7S anti-IgGs were isolated from four rabbit antistreptococcal antisera by the use of immunoabsorbent columns. The isolated proteins were of restricted molecular heterogeneity; they formed a monodisperse band on microzone electrophoresis and had a limited number of light chain bands when analyzed on urea polyacrylamide gel. The binding site of the 7S anti-IgGs was detected in the F(ab')(2) portion of the molecule. The binding site has antibody specificity for the Fc portion of IgG. For one 7S anti-IgG the combining site on the Fc portion could further be defined. A pepsin fragment of Fc, described as Pep-III', was a potent inhibitor of the coprecipitation of 7S anti-IgG with antigen-antibody complexes. An idiotypic cross-reaction was detected between the 7S and 19S anti-IgGs isolated from the same rabbit with anti-idiotype sera prepared in guinea pigs. This idiotypic specificity was not detected in the 7S anti-IgGs of 20 other rabbits.


Somatosensory evoked potentials amplitude is enhanced after non-invasive brain stimulation in chronic ischemic stroke: Preliminary results from a randomised control trial.
To investigate the effects of transcranial electrical and magnetic non-invasive brain stimulation (NIBS) protocols on somatosensory evoked potential (SEP) in chronic ischemic stroke. 33 patients were randomly assigned to one of the four treatment groups of the transcranial direct current stimulation (tDCS) and/or repetitive transcranial magnetic stimulation (rTMS) protocol. SEP parameters were recorded before and after ten days of the treatment session. All the statistical analyses were carried out using SPSS version 19. It was found that there is a statistically significant improvement in the N20-P22 mean amplitude after treatment sessions in all groups except the group where tDCS and rTMS groups were sham. On paired t-tests, the difference betweeen post and pre-stimulation SEP amplitudes for the real tDCS and real rTMS coupled group was 1.045 ± 0.732 (p value = 0.005). For sham tDCS+real rTMS group, 1.05 ± 0.96 (P = 0.04); for real tDCS+sham rTMS 0.543 ± 0.332 (P = 0.01) and for double sham stimulation, 0.204 ± 0.648 (P = 0.4) respectively CONCLUSION: In ischemic stroke patients, either or coupled true transcranial tDCS and rTMS was found to be safe and significantly enhanced the amplitude of cortical somatosensory potentials when combined with standard physiotherapy, in the interim analysis of an ongoing randomised controlled trial. CTRI/2019/11/022009 SIGNIFICANCE: The results of this research indicates the importance of RCTs in developing robust improved NIBS protocols coupled to physiotherapy to enhance the sensory-motor functional recovery following ischemic stroke.


An anatomical study of the sigmoid sinus artery: Application to the transmastoid approach.
We aimed to investigate the morphological features of the artery that traverse the sigmoid sinus's lateral surface and to discuss this structure's clinical relevance. Ten sides from five cadaveric Caucasian heads were used for gross anatomical dissection to investigate the morphological features of the sigmoid sinus artery (SSA), and additional five sides were used for histological observation. The SSA was found on eight out of ten sides (80%). The mean diameter of the SSA was 0.3 mm. The mean distance from the tip of the mastoid process to the artery was 20.3 mm. Histological observation identified extradural and intradural courses of SSA. The intradural course was further categorized into protruding and non-protruding types. In the protruding type, the SSA traveled within the dura but indented into the bone, making it more or less an intraosseous artery. In the non-protruding type, the SSA traveled within the dura but did not protrude into the bone but rather indented into the lumen of the SS. In all sections, both intradural and extradural courses were identified simultaneously. When the mastoid foramen is observed, it does not always only carry an emissary vein but also an artery. The SSA could be considered a "warning landmark" during bone drilling for the transmastoid approach.


Expression of BCL-2 via adeno-associated virus vectors rescues thalamic neurons after visual cortex lesion in the adult rat.
Lesions of the mammalian visual cortex cause the retrograde degeneration of the thalamic neurons projecting to the damaged cortex. The proto-oncogene bcl-2 is known to inhibit neuronal apoptosis induced by a variety of noxious stimuli and preserve the functional integrity of the injured cells. Here we have tested whether the overexpression of bcl-2 via adeno-associated virus (AAV) vectors is able to protect the neurons in the lateral geniculate nucleus after visual cortex ablation in adult rats. Recombinant AAV vectors encoding Bcl-2 (AAV-Bcl-2) or green fluorescent protein (AAV-GFP) as a control were stereotaxically injected into the geniculate. Three weeks after vector injection, the ipsilateral visual cortex was removed by aspiration, and cell survival was assessed 2 weeks later. We found that 20% of the geniculate neurons were transduced by the Bcl-2 vector. These cells were completely protected from death following cortical ablation. Delivery of AAV-GFP transduced an identical number of geniculate neurons but had no effect on cell survival after lesion. The total number of surviving geniculate neurons was found to be significantly higher in animals injected with AAV-Bcl-2 than in rats injected with AAV-GFP or in control lesioned rats. These data indicate that Bcl-2 gene therapy with AAV vectors represents an effective treatment to promote neuronal survival after central nervous system insults.


Epidemiological and Molecular Characterization of an Invasive Group A Streptococcus emm32.2 Outbreak.
An emm32.2 invasive group A streptococcus (iGAS) outbreak occurred in Liverpool from January 2010 to September 2012. This genotype had not previously been identified in Liverpool, but was responsible for 32% (14/44) of all iGAS cases reported during this time period. We performed a case-case comparison of emm32.2 iGAS cases with non-emm32.2 control iGAS cases identified in the Liverpool population over the same time period to assess patient risk factors for emm32.2 iGAS infection. The emm32.2 iGAS cases were confined to the adult population. We show that homelessness, intravenous drug use, and alcohol abuse predisposed patients to emm32.2 iGAS disease; however, no obvious epidemiological linkage between the patients with emm32.2 iGAS could be identified. Comparative whole-genome sequencing analysis of emm32.2 iGAS and non-emm32.2 control isolates was also performed to identify pathogen factors which might have driven the outbreak. We identified 19 genes, five of which had previously been implicated in virulence, which were present in all of the emm32.2 iGAS isolates but not present in any of the non-emm32.2 control isolates. We report that a novel emm32.2 genotype emerged in Liverpool in 2010 and identified a specific subset of genes, which could have allowed this novel emm32.2 genotype to persist in a disadvantaged population in the region over a 3-year period.


Burn Specific Health up to 24 months after the Burn-A prospective validation of the simplified model of the Burn Specific Health Scale-Brief.
Outcome after burn injury is a multidimensional concept, but few multidimensional, injury-specific outcome measures have been psychometrically evaluated. A recent cross-sectional study using the Burn Specific Health Scale-Brief (BSHS-B) found three psychometrically sound health domains: function, skin involvement, and affect and relations. The aim of this study was to reexamine the psychometric properties of the BSHS-B using a prospective study design. Ninety-four consecutive adult patients with burns were included and asked to fill in questionnaires, the BSHS-B, the Hospital Anxiety and Depression Scale, and the short-form 36 (SF-36), at 6 months, 12 months, and 24 months postburn. The factor structure was replicated and the three domains, function, skin involvement, and affect and relations, had excellent internal consistency. Over time the scores of function and skin involvement increased, indicating health improvement, whereas the domain affect and relations did not change over time. At 6 months and 12 months postburn, all domains were associated with burn severity. The function domain was highly associated with the SF-36 subscales physical functioning and role-physical, the affect and relations domain was highly associated with the Hospital Anxiety and Depression Scale and the SF-36 subscales denoting psychological health, and the domain skin involvement was highly associated with subscales indicating role-concerns, social functioning, vitality, and mental health. The psychometric properties of the BSHS-B domains were excellent and they had intelligible concurrent associations with other measures. Thus, the simplified model of the BSHS-B is a reliable, valid, and useful tool in describing postburn health over time.


[Hypocalcemia after thyroidectomy. Pathogenetic and clinical aspects].
Hypocalcaemia may arise after thyroidectomy. This condition has been attributed to various causes but the explanations of the phenomenon are not always convincing. The present study was designed to investigate the frequency of the phenomenon and its causes via the early and tardive monitoring of calcaemia, phosphoraemia, calcitoninaemia, thyroid hormones and TSH. The results obtained reveal that hypocalcaemia is due to postoperative hypothyroidism that is revealed more clearly by the behaviour of TSH than that of the thyroid hormones. Patients must be treated with vitamin D until thyroid hormone function is normalised. The behaviour of PTH after thyroidectomy and in hypothyroidism reveals the appearance of inane hyperparathyroidism secondary to and related to the hypocalcaemia.


Modeling the evolution of SIV sooty mangabey progenitor virus towards HIV-2 using humanized mice.
HIV-2 is thought to have originated from an SIV progenitor native to sooty mangabeys. To model the initial human transmission and understand the sequential viral evolution, humanized mice were infected with SIVsm and serially passaged for five generations. Productive infection was seen by week 3 during the initial challenge followed by chronic viremia and gradual CD4+ T cell decline. Viral loads increased by the 5th generation resulting in more rapid CD4+ T cell decline. Genetic analysis revealed several amino acid substitutions that were nonsynonymous and fixed in multiple hu-mice across each of the 5 generations in the nef, env and rev regions. The highest rate of substitution occurred in the nef and env regions and most were observed within the first two generations. These data demonstrated the utility of hu-mice in modeling the SIVsm transmission to the human and to evaluate its potential sequential evolution into a human pathogen of HIV-2 lineage.


Cloning and sequencing of the yeast gene for dolichol phosphate mannose synthase, an essential protein.
Dolichol phosphate mannose (Dol-P-Man) synthase (EC 126.96.36.199) catalyzes the formation of Dol-P-Man from Dol-P and GDP-Man. The structural gene for yeast Dol-P-Man synthase (DPM1) was isolated by screening a yeast genomic DNA library for colonies that overexpressed Dol-P-Man synthase activity. This approach relied on a method to screen for Dol-P-Man synthase activity in lysed yeast colonies and used a yeast mutant with very low Dol-P-Man synthase activity in colony lysates. Transformants isolated using this technique expressed Dol-P-Man synthase activity 9-14-fold higher than that of a wild type strain, and all seven plasmids conferring this overproduction had a common region in their yeast genomic DNA insert. DPM1 is the structural gene for yeast Dol-P-Man synthase since Escherichia coli transformants harboring this gene express Dol-P-Man synthase activity in vitro. DNA sequencing of the DPM1 gene revealed an open reading frame of 801 bases. The 30-kDa size of the predicted protein is in excellent agreement with the size of the purified yeast enzyme (Haselbeck, A., and Tanner, W. (1982) Proc. Natl. Acad. Sci. U. S. A. 79, 1520-1524). Analysis of the predicted amino acid sequence reveals the protein has a potential membrane spanning domain of 25 amino acids at its COOH terminus. The protein's NH2 terminus, though not hydrophobic, meets existing criteria for yeast signal sequences, but there is no site for cleavage by signal peptidase. If the NH2 terminus is a functional signal sequence, the protein is predicted to be oriented toward the lumen of the endoplasmic reticulum with both NH2 and COOH termini serving as membrane anchors. If there is no signal sequence, the enzyme is predicted to face the cytoplasm and be anchored only by its COOH terminus. The DPM1 gene is essential for viability in yeast since disruption of the gene is lethal. We suspect Dol-P-Man synthase is not an essential protein due to its role in N-glycosylation since mutations in other genes that affect the late steps in lipid-linked oligosaccharide synthesis do not affect cell growth. Instead, DPM1 may be an essential gene because its product is required for O-glycosylation in yeast or because Dol-P-Man synthase is needed in some unidentified pathway.


Abiotrophia defectiva endophthalmitis following routine cataract surgery: the first reported case in the United Kingdom.
Abiotrophia defectiva is a fastidious organism that has been implicated in severe infections such as endocarditis in immunocompetent patients. Modern tools are available to aid identification, but the main challenge remains clinical suspicion of A. defectiva . An otherwise fit and well 65-year-old female presented with reduced vision, red eye and discomfort 2 days following routine left cataract surgery. She had visual acuity of light perception only, significant anterior chamber inflammation (including hypopyon) and limited fundal view. She was diagnosed with post-operative endophthalmitis and 0.1 ml of ceftazidime (2 mg/0.1 ml) and 0.1 ml vancomycin (2 mg/0.1 ml) were injected intravitreally after vitreous aspiration. Subconjunctival cefuroxime was also injected. A repeat injection was performed on day three of admission. Gram staining revealed Gram-positive long-chain cocci, which were identified as A. defectiva . The patient was discharged on oral ciprofloxacin 500 mg twice a day with oral prednisolone 60 mg once a day; this was tapered and stopped at 8 weeks post-discharge. The left eye received dexamethasone 0.1 % 6 times a day (again, tapered over 8 weeks), moxifloxacin 5 % 6 times a day and atropine 1 % twice a day. Vision improved to 6/12 unaided (6/9.5 with pinhole) at 9 weeks post-operatively, with a clear fundal view. We present a case of A. defectiva endophthalmitis following routine cataract surgery. To our knowledge, this is the first reported case in the UK and the fourth globally, which with prompt treatment ended with a good visual outcome.


Lipopolysaccharide- and cholera toxin-specific subclass distribution of B-cell responses in cholera.
The immunoglobulin subclass responses to homologous lipopolysaccharide (LPS) and to cholera toxin (CT) in adult patients infected with Vibrio cholerae O1 and V. cholerae O139 were studied. LPS-specific antibody-secreting cells (ASC) of both the immunoglobulin A1 (IgA1) and IgA2 subclasses were seen, with the IgA1 ASC response predominating in both V. cholerae O1- and O139-infected patients. For antibodies in plasma, by day 11 after onset of disease, all V. cholerae O1- infected patients responded to homologous LPS with the IgA1 subclass (P = 0.001), whereas fewer (68%) responded with the IgA2 subclass (P = 0.007). About 89% of V. cholerae O139-infected patients responded with the IgA1 subclass (P = 0.003), and only 21% responded with the IgA2 subclass (not significant [NS]). Both groups of cholera patients showed significant increases in LPS-specific IgG1, IgG2, and IgG3 antibodies in plasma. In feces, the response to homologous LPS occurred in both groups of patients with the IgA1 and IgA2 subclasses, with 55 to 67% of patients showing a positive response. V. cholerae O1- and O139-infected patients showed CT-specific ASC responses of the different IgG and IgA subclasses in the circulation, and the pattern followed the order IgG1 > IgA1 > IgG2 > IgA2, with low levels of IgG3 and IgG4 ASC. Plasma anti-CT antibody responses in all subclasses were seen by day 11 after onset of disease. Although there were no increases in CT-specific ASC of the IgG3 (NS) and IgG4 (NS) subtypes, there were significant increases of these two subclasses in plasma (P </= 0.001). The response to CT in the fecal extracts was contributed to by both IgA1 and IgA2 isotypes, with 67 to 75% of the patients responding. Thus, the mucosa-derived ASC and fecal antibodies to LPS and CT were of both the IgA1 and IgA2 subclasses; in plasma, the contribution from IgA2 was lower. Very little difference in the B-cell responses to LPS and CT in the different subclasses was seen in the two groups of cholera patients. Vaccines against O1 and O139 cholera ideally should stimulate antibody subclasses that are likely to offer protection.


Molecular phylogeny, systematics and morphological evolution of the acorn barnacles (Thoracica: Sessilia: Balanomorpha).
The Balanomorpha are the largest group of barnacles and rank among the most diverse, commonly encountered and ecologically important marine crustaceans in the world. Paradoxically, despite their relevance and extensive study for over 150years, their evolutionary relationships are still unresolved. Classical morphological systematics was often based on non-cladistic approaches, while modern phylogenetic studies suffer from severe undersampling of taxa and characters (both molecular and morphological). Here we present a phylogenetic analysis of the familial relationships within the Balanomorpha. We estimate divergence times and examine morphological diversity based on five genes, 156 specimens, 10 fossil calibrations, and six key morphological characters. Two balanomorphan superfamilies, eight families and twelve genera were identified as polyphyletic. Chthamaloids, chionelasmatoid and pachylasmatoids split first from the pedunculated ancestors followed by a clade of tetraclitoids and coronuloids, and most of the balanoids. The Balanomorpha split from the Verrucidae (outgroup) in the Lower Cretaceous (139.6 Mya) with all the main lineages, except Pachylasmatoidea, having emerged by the Paleocene (60.9 Mya). Various degrees of convergence were observed in all the assessed morphological characters except the maxillipeds, which suggests that classical interpretations of balanomorphan morphological evolution need to be revised and reinterpreted.


[Morphology of the induction process during following bone matrix implantation in mice].
A decalcified bone matrix was implanted intramuscularly to normal mice, to mice irradiated by 450 R, to mice with curretaged femur and beneath the renal capsule in normal mice. In the course of implantation the bone induction was noted in 80% of cases. In all the series of experiments, the implants had different degrees of maturity of induced bone tissue and different extent of repopulation of the induced bone by hemopoietic cells. Hemopoietic cells inhabited young osteogenic tissue with a well marked osteoblastic layer and formed bone marrow cavity. No hemopoiesis was observed in the case of induction of cavity-free bone tissue plates or mature bones with bone marrow cavity but without osteoblastic layer.


The incidence and natural course of transfusion-associated GB virus C/hepatitis G virus infection in a cohort of thalassemic patients. The Cooleycare Cooperative Group.
To evaluate the risk of transmitting blood-borne GB virus C/hepatitis G virus (GBV-C/HGV) and to define the natural course of infection, we performed a prospective study in a cohort of multitransfused beta-thalassemics during a 6-year follow-up period. We analyzed serum samples of 150 patients collected at 3-year intervals from 1990 to 1996. GBV-C/HGV RNA was determined by reverse transcriptase-polymerase chain reaction and antibodies to E2-protein by an enzyme immunoassay. At baseline, 14.5% of patients had viremia and 18.5% anti-E2. None of the patients with anti-E2 in 1990 subsequently became viremic. Of the 100 GBV-C/HGV RNA-, anti-E2- patients, 10 acquired infection during follow-up, as indicated by positivity of GBV-C/HGV RNA (n = 2), anti-E2 (n = 7), or both markers (n = 1) in 1996. The incidence was 1.7 per 100 person-years (95% confidence interval [CI], 0.8 to 3). Since approximately 19,000 blood units were transfused to these patients during follow-up, the risk of infection was 5.3 in 10,000 units (95% CI, 2 to 8.5). Six of 22 viremic patients cleared the virus during follow-up; 4 of them became anti-E2+. Twelve of 28 patients lost anti-E2 reactivity during follow-up. In conclusion, more than 25% of infections resolve within 6 years; the presence of anti-E2 seems to be protective against infection. Anti-E2 reactivity may decrease with time.


pH dependence of the four individual transitions in the catalytic S-cycle during photosynthetic oxygen evolution.
We have investigated the pH dependence for each individual redox transition in the S-cycle of the oxygen evolving complex (OEC) of photosystem II by electron paramagnetic resonance (EPR) spectroscopy. In the experiments, OEC is advanced to the appropriate S-state at normal pH. Then, the pH is rapidly changed, and a new flash is given. The ability to advance to the next S-state in the cycle at different pHs is determined by measurements of the decrease or increase of characteristic EPR signals from the OEC in different S-states. In some cases the measured EPR signals are very small (this holds especially for the S0 ML signal at pH >7.5 and pH <4.8). Therefore, we refrain from providing error limits for the determined pK's. Our results indicate that the S1 --> S2 transition is independent of pH between 4.1 and 8.4. All other S-transitions are blocked at low pH. In the acidic region, the pK's for the inhibition of the S2 --> S3, the S3 --> [S4] --> S0, and the S0 --> S1 transitions are about 4.0, 4.5, and 4.7, respectively. The similarity of these pK values indicates that the inhibition of the steady-state oxygen evolution in the acidic range, which occurs with pK approximately 4.8, is a consequence of similar pH blocks in three of the redox steps involved in the oxygen evolution. In the alkaline region, we report a clear pH block in the S3 --> [S4] --> S0 transition with a pK of about 8.0. Our study also indicates the existence of a pH block at very high pH (pK approximately 9.4) in the S2 --> S3 transition. The S0 --> S1 transition is not affected, at least up to pH 9.0. This suggests that the inhibition of the steady-state oxygen evolution, which occurs with a pK of 8.0, is dominated by the inhibition of the S3 --> [S4] --> S0 transition. Our results are obtained in the presence of 5% methanol (v/v). However, it is unlikely that the determined pK's are affected by the presence of methanol since our results also show that the pH dependence of the steady-state oxygen evolution is not affected by methanol. The results in the alkaline region are in good agreement with a model, which suggests that the redox potential of Y(Z*)/Y(Z) is directly affected by high pH. At high pH the Y(Z*)/Y(Z) potential becomes lower than that of S2/S1 and S3/S2. The acidic block, with a pK of 4-5 in three S-transitions, implies that the inhibition mechanism is similar, and we suggest that it reflects protonation of a carboxylic side chain in the proton relay that expels protons from the OEC.


Epidemiology and natural history of atrial fibrillation: clinical implications.
With a substantial impact on morbidity and mortality, the growing "epidemic" of atrial fibrillation (AF) intersects with a number of conditions, including aging, thromboembolism, hemorrhage, hypertension and left ventricular dysfunction. Currently, the epidemiology and natural history of AF govern all aspects of its clinical management. The ongoing global investigative efforts toward understanding AF are also driven by epidemiologic findings. New developments, by affecting the natural history of the disease, could eventually alter the nature of decision making in patients with AF. The crucial issue of rate versus rhythm control awaits completion of the AF Follow-up Investigation of Rhythm Management trial. The processes of electrical and structural remodeling that perpetuate AF appear to be reversible. In the era of functional genomics, the molecular basis of this ubiquitous arrhythmia is in the process of being defined. Unraveling the molecular genetics of AF might provide new insights into the structural and electrical phenotypes resulting from genetic mutations and, as such, new approaches to treatment of this arrhythmia at the ion channel and cellular levels. Thus, current adverse trends are superimposed on a background of a rapidly developing knowledge base and potentially exciting new therapeutic options. Consequently, an understanding of the epidemiology and natural history of AF is crucial to the future allocation of resources and the utilization of an expanding range of therapies aimed at reducing the impact of this disease on a changing patient population.


Single cell activity in the auditory cortex of Rhesus monkeys: behavioral dependency.
The response to repetitive stimulation of single cells in the auditory cortex of the monkey is dependent upon behavioral performance and training of the subject in a simple auditory discrimination task. In the trained, performing animal, single cells are more responsive than in the animal that is trained but not performing in the task. In the naive monkey, evoked responses are labile and are maintained only with nonrepetitive auditory stimuli.


Physiological responses of Manila clams Venerupis (=Ruditapes) philippinarum with varying parasite Perkinsus olseni burden to toxic algal Alexandrium ostenfeldii exposure.
Manila clam stock from Arcachon Bay, France, is declining, as is commercial harvest. To understand the role of environmental biotic interactions in this decrease, effects of a toxic dinoflagellate, Alexandrium ostenfeldii, which blooms regularly in Arcachon bay, and the interaction with perkinsosis on clam physiology were investigated. Manila clams from Arcachon Bay, with variable natural levels of perkinsosis, were exposed for seven days to a mix of the nutritious microalga T-Iso and the toxic dinoflagellate A. ostenfeldii, a producer of spirolides, followed by seven days of depuration fed only T-Iso. Following sacrifice and quantification of protozoan parasite Perkinsus olseni burden, clams were divided into two groups according to intensity of the infection ("Light-Moderate" and "Moderate-Heavy"). Hemocyte and plasma responses, digestive enzyme activities, antioxidant enzyme activities in gills, and histopathological responses were analyzed. Reactive oxygen species (ROS) production in hemocytes and catalase (CAT) activity in gills increased with P. olseni intensity of infection in control clams fed T-Iso, but did not vary among A. ostenfeldii-exposed clams. Exposure to A. ostenfeldii caused tissue alterations associated with an inflammatory response and modifications in hemocyte morphology. In the gills, superoxide dismutase (SOD) activity decreased, and an increase in brown cell occurrence was seen, suggesting oxidative stress. Observations of hemocytes and brown cells in tissues during exposure and depuration suggest involvement of both cell types in detoxication processes. Results suggest that exposure to A. ostenfeldii disrupted the pro-/anti-oxidant response of clams to heavy P. olseni intensity. In addition, depressed mitochondrial membrane potential (MMP) in hemocytes of clams exposed to A. ostenfeldii suggests that mitochondrial functions are regulated to maintain homeostasis of digestive enzyme activity and condition index.


Evolving self-rated health in middle and old age: how does it differ across Black, Hispanic, and White Americans?
This research focuses on ethnic variations in the intraindividual changes in self-rated health. Data came from the Health and Retirement Study involving up to 6 repeated observations between 1995 and 2006 of a national sample of 18,486 Americans above 50 years of age. Hierarchical linear models were employed in depicting variations in self-rated health across White, Black, and Hispanic Americans. Subjective health worsened over time albeit moderately. Relative to younger persons, older individuals rated their health poorer with a greater rate of deteriorating health. With reference to ethnic variations in the intercept and slope of perceived health, White Americans rated their health most positively, followed by Black Americans, with Hispanics rating their health least positively. This pattern held even when socioeconomic status, social networks, and prior health were adjusted. Significant ethnic differences exist in the evolvement of self-rated health in middle and late life. Further inquiries may include analyzing ethnic heterogeneities from a person-centered perspective, health disparities across subgroups of Hispanics, effects of neighborhood attributes, and implications of left truncation.


[Immunoleukolysis test results in women immunized with staphylococcal toxoid].
The results of the immunoleukolysis test in 699 women on the 32nd week of pregnancy are presented. Of these, 48 women were examined 1 month and 6 months after the third injection. In this test staphylococcal allergen and E. coli allergen were used. Staphylococcal toxoid was found to have desensitizing action in persons with enhanced initial allergic state. The authors conclude that the immunoleukolysis test may ensure differential approach to the immunization of pregnant women with staphylococcal toxoid.


Clinical manifestations and outcome of patients with primary amoebic meningoencephalitis in Pakistan. A single-center experience.
Primary amoebic meningoencephalitis (PAM) is a rapidly progressing central nervous system (CNS) infection caused by Naegleria fowleri, a free-living amoeba found in warm freshwater. The disease progression is very rapid, and the outcome is nearly always fatal. We aim to describe the disease course in patients admitted with PAM in a tertiary care center in Karachi, Pakistan between the periods of 2010 to 2021. A total of 39 patients were included in the study, 33 males (84.6%). The median age of the patients was 34 years. The most frequent presenting complaint was fever, which was found in 37 patients (94.9%) followed by headache in 28 patients (71.8%), nausea and vomiting in 27 patients (69.2%), and seizures in 10 patients (25.6%). Overall, 39 patients underwent lumbar puncture, 27 patients (69.2%) had a positive motile trophozoites on CSF wet preparation microscopy, 18 patients (46.2%) had a positive culture, and 10 patients had a positive PCR. CSF analysis resembled bacterial meningitis with elevated white blood cell counts with predominantly neutrophils (median, 3000 [range, 1350-7500] cells/μL), low glucose levels median, 14 [range, 1-92] mg/dL), and elevated protein levels (median, 344 [range, 289-405] mg/dL). Imaging results were abnormal in approximately three-fourths of the patients which included cerebral edema (66.7%), hydrocephalus (25.6%), and cerebral infarctions (12.8%). Only one patient survived. PAM is a fatal illness with limited treatment success. Early diagnosis and prompt initiation of treatment can improve the survival of the patients and reduce mortality.


Hippocampal head atrophy predominance in Parkinson's disease with hallucinations and with dementia.
We studied regional gray matter density in the hippocampus in Parkinson's disease (PD) patients. We obtained magnetic resonance scans in 44 PD patients (PD patients with dementia (PDD) = 9, non-demented PD patients with visual hallucinations (PD + VH) = 16, and PD patients without dementia and without visual hallucinations (PD - VH) = 19) and 56 controls matched for age and years of education. A region of interest (ROI) of the hippocampus following voxel-based morphometry (VBM) procedures was used to perform group comparisons, single-case individual analysis and correlations with learning scores. Group comparisons showed that PDD patients and PD+VH patients had significant hippocampal gray matter loss compared to controls. In PDD patients, hippocampal gray matter loss involved the entire hippocampus and in PD+VH this reduction was mainly confined to the hippocampal head. 78 % of PDD patients, 31 % of PD+VH patients and 26 % of PD-VH patients had hippocampal head gray matter loss when compared to controls. These results suggest that in PD the neurodegenerative process in the hippocampus starts in the head of this structure and later spreads to the tail and that, in addition, memory impairment assessed by Rey's Auditory Verbal Learning Test (RAVLT) correlates with hippocampal head gray matter loss.


Effects of noncompetitive NMDA receptor blockade on anterior cingulate cerebral blood flow in volunteers with schizophrenia.
Schizophrenia may be related to dysfunctional glutamatergic activity, specifically hypofunction of the N-methyl-D-aspartate receptor (NMDAR). In addition, it has been proposed that NMDAR hypofunction may paradoxically cause an increase in glutamate release and hypermetabolism in corticolimbic regions. If a state of partial, chronic NMDAR blockade underlies schizophrenia, then schizophrenic volunteers (SV) may have greater glutamate release and associated elevations in regional cerebral blood flow (rCBF) than normal volunteers (NV), following drug-induced NMDAR antagonism. Therefore, we have given acute ketamine, a noncompetitive NMDAR antagonist, to NV (n=13) and medicated volunteers with schizophrenia (n=10) in conjunction with serial positron emission tomography blood flow studies. Drug administration caused marked rCBF elevations in frontal and cingulate regions in both groups. Contrasts between NV and SV ketamine groups showed that SV had greater relative blood flow increases in the anterior cingulate than NV. Maximum blood flow, and the area under the curve for blood flow in the anterior cingulate cortex, significantly correlated with changes in psychosis ratings in SV and NV (maximum rCBF only). These changes are consistent with a relatively hypoactive thalamic NMDAR and increased cortical glutamate neurotransmission at non-NMDARs in schizophrenia. We hypothesize that ketamine antagonizes an NMDAR-dependent inhibitory system that is partially compromised in subjects with schizophrenia. The ketamine-induced reduction of inhibition leads to a marked increase in glutamate release and hypermetabolism (elevated rCBF) in frontal and cingulate cortical regions. The loss of inhibition and increased glutamate release may cause the distorted thoughts and diminished cognitive abilities elicited by NMDAR blockade.


Working memory capacity predicts focus back effort under different task demands.
According to the cognitive flexibility view, individuals with higher cognitive control ability are more flexible in experiencing on task or mind wandering during tasks with different loads. On the other hand, the resource-control theory posits that executive control is essential for allocating attentional resources between mind wandering and tasks. Focus back effort may reflect the adjustment of executive control in the resource-control theory. Here, 121 participants completed two span tasks, as well as high- and low-load tasks, while mind wandering and focus back effort were measured. Our findings indicated that mind wandering was influenced by working memory capacity (WMC) and focus back effort. Additionally, participants demonstrated a higher focus back effort during the higher load task. This effect was particularly pronounced in individuals with lower WMC, which was treated as a continuous variable. These findings integrate the cognitive flexibility view and resource-control theory to describe how individuals modulate mind wandering.


Effect of metoclopramide given before atropine sulphate on lower oesophageal sphincter tone.
The effects on lower oesophageal sphincter (LOS) tone of sequential intravenous injections of metoclopramide 10 mg/atropine 0,6 mg and atropine 0,6 mg/metoclopramide 10 mg, given randomly on separate occasions to 8 healthy volunteers, were studied. The administration of metoclopramide increased mean LOS pressure by 13,6 cm H2O (P less than 0,001). Subsequent injection of atropine failed to lower LOS pressure significantly, LOS pressure being sustained at a mean of 11,2 cm H2O above basal control levels (P less than 0,01). In contrast, injection of atropine at a later date in the same subjects lowered the average LOS pressure by 10,5 cm H2O (P less than 0,001), and subsequent intravenous injections of metoclopramide restored LOS pressure to basal levels. The results of this study suggest that metoclopramide should be given prior to atropine before induction of general anaesthesia to counteract the deleterious effects of atropine on LOS tone, thereby helping to reduce the chances of regurgitation and pulmonary aspiration of acid gastric contents.


Tissue-specificity, functional characterization and subcellular localization of a rat ubiquitin-specific processing protease, UBP109, whose mRNA expression is developmentally regulated.
A cDNA encoding an ubiquitin-specific processing protease, UBP109, in rat skeletal muscle was cloned and its product was characterized. Northern analysis revealed that UBP109 mRNA is highly expressed in testis and spleen, compared with other tissues. Furthermore, in situ hybridization showed that the level of UBP109 mRNA in liver, spinal cord and brain dramatically changed during embryonic development, indicating that the expression of UBP109 mRNA is developmentally regulated. UBP109 was expressed in Escherichia coli and purified to apparent homogeneity using a (125)I-labelled ubiquitin-peptide fusion as a substrate. The purified enzyme cleaved at the C-terminus of the ubiquitin moiety in natural and engineered fusions irrespective of their sizes. UBP109 also released free ubiquitin from poly-His-tagged penta-ubiquitin. Moreover, it released free ubiquitin from poly-ubiquitinated protein conjugates of rabbit reticulocytes. In addition, UBP109 localized to both the cytoplasm and the nucleus and, among three putative nuclear localization sequences, only the one located near the C-terminus is responsible for nuclear localization. These results suggest that UBP109 may play an important role in generation of free ubiquitin from its precursors and its recycling from poly-ubiquitinated protein conjugates, and hence in regulation of ubiquitin-mediated cellular processes, particularly related to embryonic development.


Gamma-secretase inhibitor treatment promotes VEGF-A-driven blood vessel growth and vascular leakage but disrupts neovascular perfusion.
The Notch signaling pathway is essential for normal development due to its role in control of cell differentiation, proliferation and survival. It is also critically involved in tumorigenesis and cancer progression. A key enzyme in the activation of Notch signaling is the gamma-secretase protein complex and therefore, gamma-secretase inhibitors (GSIs)--originally developed for Alzheimer's disease--are now being evaluated in clinical trials for human malignancies. It is also clear that Notch plays an important role in angiogenesis driven by Vascular Endothelial Growth Factor A (VEGF-A)--a process instrumental for tumor growth and metastasis. The effect of GSIs on tumor vasculature has not been conclusively determined. Here we report that Compound X (CX), a GSI previously reported to potently inhibit Notch signaling in vitro and in vivo, promotes angiogenic sprouting in vitro and during developmental angiogenesis in mice. Furthermore, CX treatment suppresses tumor growth in a mouse model of renal carcinoma, leads to the formation of abnormal vessels and an increased tumor vascular density. Using a rabbit model of VEGF-A-driven angiogenesis in skeletal muscle, we demonstrate that CX treatment promotes abnormal blood vessel growth characterized by vessel occlusion, disrupted blood flow, and increased vascular leakage. Based on these findings, we propose a model for how GSIs and other Notch inhibitors disrupt tumor blood vessel perfusion, which might be useful for understanding this new class of anti-cancer agents.


Omovertebral bone causing traumatic compression of the cervical spinal cord and acute neurological deficits in a patient with Sprengel's deformity and Klippel-Feil syndrome: case report.
The presence of an omovertebral bone with Sprengel's deformity and Klippel-Feil syndrome is a complex congenital anomaly that is not well understood. It most commonly manifests as cosmetic deformity, limited range of motion, and functional disability, although there are reports of the insidious development of cervical myelopathy. In this paper, the authors present the case of a 49-year-old man with acute neurological deficits after a low-energy mechanism of traumatic spinal cord compression, resulting from an impinging omovertebral bone through a traumatic laminar defect. The patient underwent resection of the omovertebral bone, laminectomy decompression of the spinal canal, and anterior stabilization. This case highlights a rarely discussed complication of undiagnosed Sprengel's deformity and its associated conditions following even low-energy traumatic mechanisms.


A CONSORT-compliant prospective randomized controlled trial: radiation dose reducing in computed tomography using an additional lateral scout view combined with automatic tube current modulation: Phantom and patient study.
Radiation exposure has been a hot point in research field of computed tomography (CT). Recently, automated tube current modulation (ATCM) has emerged as an important technique to reduce radiation exposure. Many studies have shown that the difference in scout view would affect modulation. This prospective randomized controlled study is aimed to investigate the impact of an additional lateral scout view on radiation dose and image quality in CT using ACTM. Combined with ATCM (Care Dose 4D) on multidetector CT, 2 thoracic phantom CT image series were acquired in which planning was conducted with either an anteroposterior (AP) or an AP-lateral scout view. Also, 410 patients underwent thoracic CT examinations using Care Dose 4D modulation and were randomized to either a scan planned with an AP-lateral scout or a single AP scout. Effects of the different scout views on applied effective milliampere seconds (mAs), volume CT dose index (CTDIvol) and dose-length-product (DLP) were analyzed. The quality of patient CT images was also assessed. Data were analyzed using independent t tests and linear correlation analysis. Compared with AP groups, the mean CTDIvol (phantom, 0.89 ± 0.08 vs 1.36 ± 0.26 mGy, P < .001; in patients, 1.12 [0.96, 1.34] vs 2.16 [1.66, 2.64] mGy, P < .001) and DLP (in phantom, 26 [23.25, 28] vs 40 [34.25, 48] mGy×cm, P < .001; in patients, 41 [33, 41] vs 77 [60.5, 99.5] mGy×cm, P < .001) were significantly reduced by approximately 50% in AP-lateral scout view group. With the AP-lateral topogram, the radiation dose on different off-center positions was essentially equal (CTDIvol: 0.76-0.99 mGy; DLP: 22-28 mGy×cm effective dose: 0.31-0. 39 mSv). For image quality, contrast-to-noise ratio and signal-to-noise ratio values in the AP group were similar to those of AP-lateral scout view group. AP combined with an additional lateral scout view using ACTM can significantly reduce the radiation dose without compromising image quality in chest screening CT.


Comparison of partially attended night time respiratory recordings and full polysomnography in patients with suspected sleep apnoea/hypopnoea syndrome.
Laboratory full polysomnography (PSG) is considered to be the gold standard for the diagnosis of the sleep apnoea/hypopnoea syndrome (SAHS), but it is expensive and time consuming. A study was undertaken to evaluate the diagnostic usefulness of a partially attended night time respiratory recording (NTRR) and a clinical questionnaire in patients with suspected SAHS in comparison with full PSG. Seventy six patients (54 men) of mean (SD) age 51 (11.5) years with a body mass index of 31 (5.7) kg/m2 were studied at random on two different nights with full PSG at the sleep laboratory and with NTRR on a respiratory ward. NTRR records oximetry, airflow, chest and abdominal motion. All signals were continuously displayed on a computer screen throughout the night and respiratory events were scored automatically the following morning. All patients completed a clinical questionnaire. Mean values of the apnoea/hypopnoea index (AHI) using NTRR were lower than those obtained with full PSG (22.7 (2.4) versus 32.2 (3) events/hour) which was mainly due to underrecognition of hypopnoeas. Sensitivity and specificity of NTRR for the diagnosis of SAHS were 82% and 90%, respectively, taking as reference AHI > 10 on full PSG (AHI-PSG > 10). The mean (+/-2SD) difference in AHI between the two methods was 9.6 (range -5.4-24.6) (95% confidence interval 6.2 to 13). Symptoms of witnessed apnoeas, impotence, the overall clinical impression of a trained physician, and a neck size over 40 cm were significantly more prevalent in patients with AHI-PSG of > 10, but impotence was the only clinical feature significantly more prevalent in patients with false negative compared with true negative NTRR results that helped to distinguish patients with NTRR < 10 but AHI-PSG > 10. NTRR is a helpful and easy complementary diagnostic tool in clinical practice because it detects patients with moderate to severe SAHS reasonably well and therefore can be useful for confirming a diagnosis of SAHS and also for treatment decisions. It is suggested that patients with suspicion of SAHS should be initially studied by NTRR. When NTRR is negative, a full PSG should be performed if witnessed apnoeas, impotence, systemic hypertension, ischaemic heart disease, and a trained physician's clinical impression of SAHS are present.


Adenomyosis and obstetric complications: A retrospective case-control study.
Adenomyosis is a uterine pathology affecting an increasing number of women of childbearing age. Its diagnosis is based upon histology or imaging [ultrasound or magnetic resonance imaging (MRI)]. Several studies have investigated the impact of adenomyosis on obstetric complications, with its diagnosis based on clinical symptoms, ultrasound or composite criteria. The aim of this study was to identify potential obstetric complications related to adenomyosis in women with an MRI-confirmed diagnosis. A single centre retrospective case-control study was undertaken in pregnant patients with an MRI-confirmed diagnosis of adenomyosis between January 2013 and December 2017 at the University Hospitals of Strasbourg. Controls were matched in a 4:1 ratio for age, parity and body mass index. Multivariate analysis was performed to identify obstetric complications. In total, 291 women with an MRI-confirmed diagnosis of adenomyosis were identified during the study period. Of these, 89 patients achieved pregnancy after 24 weeks of gestation. The mean age of patients was 30.8 years. The adenomyosis group and the control group were comparable for matching criteria. Adenomyosis was found to be associated with increased risk of caesarean section [odds ratio (OR) 1.1, 95 % confidence interval (CI) 1.0-1.2; p = 0.03], intrauterine growth restriction (OR 1.3, 95 % CI 1.1-1.4; p < 0.001), postpartum haemorrhage (OR 1.2, 95 % CI 1.1- 1.4; p < 0.01), pre-eclampsia (OR 1.3, 95 % CI 1.0-1.6; p = 0.004) and previous spontaneous miscarriage (OR 2.09, 95 % CI 1.36-3.33; p < 0.001). Premature rupture of membranes, preterm delivery, severe intrauterine growth restriction and the risk of placenta praevia were not significantly higher in the adenomyosis group compared with the control group on multivariate analysis. This study demonstrates increased risk of several obstetric complications (caesarean section, intrauterine growth restriction, postpartum haemorrhage, pre-eclampsia, history of spontaneous miscarriage) in women with adenomyosis. To the authors' knowledge, this is the first study to use MRI as the sole criterion for diagnosis. These results could be complemented by larger-scale prospective studies in order to manage these patients more effectively during pregnancy.


The impact of community workshops on improving early childhood oral health knowledge.
The purpose of this study was to evaluate the effectiveness of community workshops designed to equip participants with early childhood oral health (ECOH) knowledge and early childhood caries (ECC) prevention. Convenience sample of individuals working with infants and preschool children attending an ECOH training workshop completed a questionnaire before the workshop. One month later, participants completed a follow-up questionnaire. A P-value </=.05 denoted significance. One hundred eight participants from southern Manitoba, Canada, completed the initial survey, while 67% completed the postworkshop questionnaire. Initially, many were unfamiliar with the recommended age of a first dental visit, assessing caries-risk, and identifying early stages of decay. Following the workshop, there was a 16% increase in the proportion of correct answers and a significant improvement in the number of correct choices (P<.01). Some questions showing considerable improvement included: when children should first visit the dentist (P<.001); mother having active decay placing their infant at high risk for caries (P<.001); and age until caregivers should supervise tooth-brushing (P<.001). Self-reported data suggests participants changed behaviors as a result of what they learned. Capacity-building workshops increased oral health knowledge and self-reported behaviors. This provides support that nondental professionals can effectively provide oral health education.


Elevated serum levels of mucin-associated antigen in patients with acute respiratory distress syndrome.
Increased serum levels of mucin-associated antigen have been previously demonstrated in patients with cystic fibrosis (CF) and interstitial pneumonia, and in lung-transplant recipients. The present study assessed the serum airway mucin levels in patients with acute respiratory distress syndrome (ARDS). An enzyme-linked immunosorbent assay (ELISA) method with a human-airway-mucin-specific monoclonal antibody (17Q2) was used to measure serum mucin levels in normal subjects, chronic smokers, patients with chronic bronchitis and other pulmonary diseases, patients with acute cardiogenic lung edema, and patients with ARDS. The serum mucin levels measured 9.9 +/- 0.8 ng/ml (mean +/- SEM, n = 59) in normal subjects, 12.7 +/- 1.6 ng/ml (n = 29) in chronic smokers, 21.8 +/- 1.9 ng/ml (n = 28) in patients with chronic bronchitis and other pulmonary diseases, 9.0 +/- 3.1 ng/ml (n = 5) in patients with acute cardiogenic lung edema. The serum mucin level was 53.8 +/- 6.6 ng/ml (n = 13) in patients with ARDS (p < 0.05, as compared with the four other groups). Serial measurements of serum mucin levels were obtained in patients with ARDS. Statistical analysis showed an inverse correlation of serial measurements of serum mucin with static respiratory-system compliance (p = 0.021), an inverse correlation of sequential serum mucin levels and log(Pa(O2)/Fl(O2)) (p = 0.016), and a positive correlation of sequential serum mucin levels and lung injury score (LIS) (p = 0.019). Gel-filtration analysis showed that mucin-associated antigens in ARDS sera were polydispersed and smaller than the antigens in normal sera. This study indicates that an increasing amount of degraded mucin occurs in patients with ARDS.


A social network of hospital acquired infection built from electronic medical record data.
Social networks have been used in the study of outbreaks of infectious diseases, including in small group settings such as individual hospitals. Collecting the data needed to create such networks, however, can be time consuming, costly, and error prone. We sought to create a social network of hospital inpatients using electronic medical record (EMR) data already collected for other purposes, for use in simulating outbreaks of nosocomial infections. We used the EMR data warehouse of a tertiary academic hospital to model contact among inpatients. Patient-to-patient contact due to shared rooms was inferred from admission-discharge-transfer data, and contact with healthcare workers was inferred from clinical documents. Contacts were used to generate a social network, which was then used to conduct probabilistic simulations of nosocomial outbreaks of methicillin-resistant Staphylococcus aureus and influenza. Simulations of infection transmission across the network reflected the staffing and patient flow practices of the hospital. Simulations modeling patient isolation, increased hand hygiene, and staff vaccination showed a decrease in the spread of infection. We developed a method of generating a social network of hospital inpatients from EMR data. This method allows the derivation of networks that reflect the local hospital environment, obviate the need for simulated or manually collected data, and can be updated in near real time. Inpatient social networks represent a novel secondary use of EMR data, and can be used to simulate nosocomial infections. Future work should focus on prospective validation of the simulations, and adapting such networks to other tasks.


Perineal colostomy: an alternative to avoid permanent abdominal colostomy: operative technique, results and reflection.
The most common injury to indicate definitive stoma is rectal cancer. Despite advances in surgical treatment, the abdominoperineal resection is still the most effective operation in radical treatment of malignancies of the distal rectum invading the sphincter and anal canal. Even with all the effort that surgeons have to preserve anal sphincters, abdominoperineal amputation is still indicated, and a definitive abdominal colostomy is necessary. This surgery requires patients to live with a definitive abdominal colostomy, which is a condition that modify body image, is not without morbidity and has great impact on the quality of life. To evaluate the technique of abdominoperineal amputation with perineal colostomy with irrigation as an alternative to permanent abdominal colostomy. Retrospective analysis of medical records of 55 patients underwent abdominoperineal resection of the rectum with perineal colostomy in the period 1989-2010. The mean age was 58 years, 40 % men and 60 % women. In 94.5% of patients the indication for surgery was for cancer of the rectum. In some patients were made three valves, other two valves and in the remaining no valve at all. Complications were: mucosal prolapse, necrosis of the lowered segment and stenosis. The abdominoperineal amputation with perineal colostomy is a good therapeutic option in the armamentarium of the surgical treatment of rectal cancer.


The karyopherin Kap122p/Pdr6p imports both subunits of the transcription factor IIA into the nucleus.
We discovered a nuclear import pathway mediated by the product of the previously identified Saccharomyces cerevisiae gene PDR6 (pleiotropic drug resistance). This gene product functions as a karyopherin (Kap) for nuclear import. Consistent with previously proposed nomenclature, we have renamed this gene KAP122. Kap122p was localized both to the cytoplasm and the nucleus. As a prominent import substrate of Kap122p, we identified the complex of the large and small subunit (Toa1p and Toa2p, respectively) of the general transcription factor IIA (TFIIA). Recombinant GST-Kap122p formed a complex with recombinant His(6)-Toa1p/Toa2p. In wild-type cells, Toa1p and Toa2p were localized to the nucleus. Consistent with Kap122p being the principal Kap for import of the Toa1p-Toa2p complex, we found that deletion of KAP122 results in increased cytoplasmic localization of both Toa1p and Toa2p. Deletion of KAP122 is not lethal, although deletion of TOA1 and TOA2 is. Together these data suggest that Kap122p is the major Kap for the import of Toa1p-Toa2p into the nucleus. Like other substrate-Kap complexes, the Toa1p/Toa2p/Kap122p complex isolated from yeast cytosol or reconstituted from recombinant proteins, was dissociated by RanGTP but not RanGDP. Kap122p bound to nucleoporins, specifically, to the peptide repeat-containing fragments of Nup1p and Nup2p.


Neutrophil-to-lymphocyte ratio may be associated with the outcome in patients with prostate cancer.
Evidences have shown that neutrophil-to-lymphocyte ratio (NLR) has a prognostic value in patients with cancer. We wanted to test the prognostic significance of NLR in prostatic cancer of patients who are candidate to radical prostatectomy. We have considered 731 patients. Complete demographic data including age, tumor stage, Gleason score, complete blood count and serum biochemical profile were collected. Pre-treatment percentage of neutrophils and NLR were considered, and correlated with patients data and recurrence free survival. 389 patients were evaluated, mean age 65 years, mean follow-up 51.5 months, mean recurrence free survival 51.3 months. Total neutrophil count does not correlate with biochemical recurrence and disease free survival. Patients with a value higher of 60% of neutrophils are more likely to have a recurrence. Patients with a total lymphocyte count <1,500 have a higher rate of relapse. NLR was not correlated with baseline total PSA, with Gleason score and with pathological stage; patients with a NLR >3 has a higher incidence of recurrence. In multivariate analysis including age, total PSA and NLR, NLR is the most important factor able to predict recurrence. There are some limitations to this study; first, this is a retrospective study, and the total number of patients analyzed is relatively small. Our study suggests that pre-treatment NLR may be associated with disease free survival in patients with prostate cancer, and could be introduced in clinical practice. NLR has the advantage of low economic cost and wide availability.


Potent Fungal Chitinase for the Bioconversion of Mycelial Waste.
Aspergillus niger mycelial waste is a good raw material for production of N-acetyl-d-glucosamine (GlcNAc). In this study, AnChiB, an A. niger chitinase which is upregulated during autolysis, was found to degrade A. niger mycelial waste with high efficiency. It could produce 1.45 mM (GlcNAc)2 in 8 h from raw mycelial waste, outperforming other chitinases, including bacterial SmChiA, human HsCht, and insect OfChtI and OfChi-h. The crystal structure of AnChiB was determined, and residues Trp106 and Trp118 were found to be important for the activity of AnChiB toward mycelial waste; mutation of either Trp106 or Trp118 into phenylalanine or alanine resulted in dramatically decreased activity. A recombinant strain of Bacillus subtilis was constructed to extracellularly produce AnChiB, and the culture supernatant was used to treat mycelial waste. This eco-friendly strategy could produce 3.7 mM of GlcNAc from 10 g of mycelial waste in 94 h with a yield of 71.3%.


Clinicopathological Study of 30 Cases of Peripheral T-cell Lymphoma with Hodgkin and Reed-Sternberg-like B-cells from Japan.
The presence of Hodgkin and Reed-Sternberg (HRS)-like B-cells in peripheral T-cell lymphoma (PTCL) is rare and its clinicopathological features still remain unclear. Here, we describe 30 cases of PTCL with HRS-like B-cells from Japan. Twenty-three cases (77%) presented evidence of follicular T-helper phenotype (TFH) derivation: 12 were angioimmunoblastic T-cell lymphoma and 11 PTCL with TFH phenotype (PTCL-TFH). The remaining seven cases were diagnosed as PTCL, not otherwise specified (PTCL-NOS). Epstein-Barr virus (EBV) reactivation was detected in 25 cases (83%), but HRS-like B-cells were EBER in only 20 cases (67%). The median age at diagnosis was 77 years (range, 39-91 y), including 24 patients (80%) were older than 60 years of age. Most of the patients presented at an advanced clinical stage and were associated with higher risk according to the International Prognostic Index. The 3-year overall and progression-free survival rates were 44% and 27%, respectively. No significant clinicopathological differences were detected between PTCL-TFH, PTCL-NOS and the angioimmunoblastic cases. Cases with EBER HRS-like B-cells were associated with inferior overall and progression-free survival compared to those with EBER HRS-like B-cells, but the difference was not significant. In conclusion, HRS-like B-cells were found in a subset of T-cell lymphomas, especially in association with the TFH phenotype and EBV reactivation. These cells have a tendency to affect elderly patients and to be associated with advanced clinical stages and dismal prognosis. The EBV status of HRS-like B-cells does not seem to affect the clinicopathological features of this group of PTCLs.


The evolving role of systemic therapy in high risk prostate cancer: strategies for cure in the 21st century.
High-risk prostate cancer is a heterogeneous group that includes patients with clinically locally advanced stage disease at diagnosis. Unlike overt locally advanced disease, prediction of risk in clinically localized disease at an individual patient level, is not always easy or accurate with present knowledge. Gleason score, pretreatment prostate specific antigen (PSA), and stage (capsular invasion, seminal vesicle and nodal involvement) are the universally recognized criteria used to define risk. Overall, this group of patients have a greater than 50% risk of relapse. Historically, local treatment modalities with radical prostatectomy or radiation therapy constituted the mainstay of therapy in the majority of localized prostate cancer patients. However, the primary cause of failure and disease mortality stems from the development of systemic metastases. As we continue to witness stage migration towards earlier stage disease (presumably PSA related) and mortality reduction, devising better strategies for cure is a must. Recently completed randomized trials indicate a benefit from the use of hormonal therapy in patients with locally advanced prostate cancer treated with radiation therapy or node positive patients, post radical prostatectomy. While hormone-based combined modality trials have consistently shown improvements in local and systemic disease control, only two of these demonstrated improvements in overall survival. The palliative benefit of chemotherapy in hormone refractory disease and the promising response rates with newer agents has evoked interest in the use of chemotherapy in high-risk prostate cancer in the adjuvant and neoadjuvant settings. Several phase II and III trials are ongoing. Novel avenues of therapy such as tyrosine kinase inhibitors, gene therapy and angiogenesis inhibitors incorporated in a multimodality treatment strategy are likely to impact the course of this disease in the future.


Evaluation of the Interplay between Uptake Transport and CYP3A4 Induction in Micropatterned Cocultured Hepatocytes.
Previously we assessed the inductive response of prototypical inducers in hepatocyte monocultures and the long-term coculture model HepatoPac using cryopreserved hepatocytes from the same donors. We noted that the rifampicin EC50 generated using the HepatoPac model corresponded better to the EC50 based on clinical data compared with data generated in the monoculture system. We postulated that there may be differences in the functioning of uptake transporters between the two systems that may have led to the EC50 difference. In this study, we characterized the functional activity of multiple uptake transporters in the two systems using cryopreserved hepatocytes from the same donors. Our data suggest that uptake transporter activity is higher in HepatoPac compared with the monoculture system. As a follow up to this study, we measured the intracellular concentrations of rifampicin and bosentan, which are known substrates of uptake transporters; we observed significantly higher intracellular concentrations of both compounds in HepatoPac relative to the monoculture system. This finding equated to lower cytochrome P450 isoform 3A4 (CYP3A4) EC50 values in the HepatoPac system compared with the monoculture system for both mRNA and activity. In parallel, no significant EC50 shift was observed for carbamazepine and phenytoin, which are not known to be substrates of uptake transporters. Our data suggest that next generation liver models such as HepatoPac may be a useful in vitro tool to quantitatively predict drug-drug interactions when it is known that the perpetrator is also a substrate of drug transporters.


Effects of adlay on azoxymethane-induced colon carcinogenesis in rats.
Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) is a grass crop used in traditional Chinese medicine and as a nutritious food. It has been reported that adlay has anti-inflammatory and anti-tumor activity. Cyclooxygenase-2 (COX-2) is an inducible enzyme functionally related to both inflammation and colon carcinogenesis and is the target of many chemopreventive agents. This study investigated the effect of adlay on colon carcinogenesis and COX-2 expression. In a short-term experiment, male F344 rats were fed diets containing different doses of dehulled adlay and received the colon-specific carcinogen, azoxymethane (AOM), by intraperitoneal injection. All rats were killed after 5 weeks of feeding, and the colons were examined for the preneoplastic lesion, aberrant crypt foci (ACF). Dietary dehulled adlay at levels of 10%, 20%, or 40% significantly reduced the numbers of ACF and aberrant crypts. Dehulled adlay reduced the number of ACF of different sizes but did not affect the crypt multiplicity. Most ACF were found in the middle and distal colons; dehulled adlay significantly suppressed the formation of ACF in the middle colon. In a long-term experiment, male F344 rats were fed diets containing different doses of dehulled adlay and injected with AOM. All rats were killed after 52 weeks of feeding, and colons were examined for tumors and COX-2 protein expression. The results indicated that dehulled adlay did not inhibit colon tumors in spite of a slight suppressing effect in the proximal colon. Rats fed diets containing 20% dehulled adlay had less COX-2 protein expression in both proximal and distal colon tumors. The inconsistent effects between COX-2 protein expression and tumor outcome may be due to regional differences in the colon and the malignancy of the tumors. These findings suggest that dehulled adlay suppresses early events in colon carcinogenesis but not the formation of tumors.


Multisite phosphorylation provides a reliable mechanism for making decisions in noisy environments.
The ability to make decisions at the cellular level is absolutely critical for the survival of organisms. Eukaryotic cells are constantly making binary decisions in response to internal and environmental signals. Among the most notable transducers of information are protein kinases. The regulation of these signaling proteins often relies on the activity of other protein kinases located upstream in the signaling cascade. However, these signaling systems are by their own nature an important source of molecular noise. Herein, we have assessed the role of multisite phosphorylation in detecting signals in the face of molecular noise. To address this issue, we have conceptually envisioned the biochemical transduction machinery as a classifier model that can lead to four possible outputs: true positives and negatives, and false positives and negatives. In this probabilistic framework, we show that multisite phosphorylation represents a mechanism to filter noise during the decision-making process. We present results showing that nonessential phosphorylation sites contribute to increase the rate of true positives while, at the same time, they can lessen the rate of false positives. This simultaneous increase in sensitivity and specificity, makes multisite phosphorylation a valuable and easily implemented mechanism to reliably transduce information in noisy contexts.


A simple convolutional neural network for prediction of enhancer-promoter interactions with DNA sequence data.
Enhancer-promoter interactions (EPIs) in the genome play an important role in transcriptional regulation. EPIs can be useful in boosting statistical power and enhancing mechanistic interpretation for disease- or trait-associated genetic variants in genome-wide association studies. Instead of expensive and time-consuming biological experiments, computational prediction of EPIs with DNA sequence and other genomic data is a fast and viable alternative. In particular, deep learning and other machine learning methods have been demonstrated with promising performance. First, using a published human cell line dataset, we demonstrate that a simple convolutional neural network (CNN) performs as well as, if no better than, a more complicated and state-of-the-art architecture, a hybrid of a CNN and a recurrent neural network. More importantly, in spite of the well-known cell line-specific EPIs (and corresponding gene expression), in contrast to the standard practice of training and predicting for each cell line separately, we propose two transfer learning approaches to training a model using all cell lines to various extents, leading to substantially improved predictive performance. Computer code is available at https://github.com/zzUMN/Combine-CNN-Enhancer-and-Promoters. Supplementary data are available at Bioinformatics online.


ExACtly zero or once: A clinically helpful guide to assessing genetic variants in mild epilepsies.
To assist the interpretation of genomic data for common epilepsies, we asked whether variants implicated in mild epilepsies in autosomal dominant families are present in the general population. We studied 12 genes for the milder epilepsies and identified published variants with strong segregation support (de novo germline mutation or ≥4 affected family members). These variants were checked in the Exome Aggregation Consortium (ExAC), a database of genetic variation in over 60,000 individuals. We subsequently evaluated variants in these epilepsy genes that lacked strong segregation support. To determine whether the findings in epilepsies were representative of other diseases, we also assessed the presence of variants in other dominant neurologic disorders (e.g., CADASIL). Published epilepsy variants with strong segregation support (n = 65) were absent (n = 61) or present once (n = 4) in ExAC. By contrast, of 46 published epilepsy variants without strong segregation support, 8 occurred recurrently (2-186 times). Similarly, none of the 45 disease-associated variants from other neurologic disorders with strong segregation support occurred more than once in ExAC. Reanalysis using the larger ExAC V2 plus gnomAD reference cohort showed consistent results. Variants causing autosomal dominant epilepsies are ultra-rare in the general population. Variants observed more than once in ExAC were only found among reports without strong segregation support, suggesting that they may be benign. Clinicians are increasingly faced with the interpretation of genetic variants of unknown significance. These data illustrate that variants present more than once in ExAC are less likely to be pathogenic, reinforcing the valuable clinical role of ExAC.


Interaction of both the C2A and C2B domains of rabphilin3 with Ca2+ and phospholipid.
Rabphilin3, a downstream target molecule of the Rab3 subfamily small G proteins, has two C2 domains (the C2A and C2B domains) at its C-terminal region and is implicated in Ca(2+)-dependent neurotransmitter release. Rabphilin3 interacts with Ca2+ and phospholipid at its C2 domain, but it remains to be clarified which domain, the C2A or C2B domain, interacts with these compounds. We have found here that both the C2A and C2B domains interact with Ca2+ with similar kinetics but interact with phospholipid with slightly different kinetics.


Brønsted Acid-Catalyzed Synthesis of 4-Functionalized Tetrahydrocarbazol-1-ones from 1,4-Dicarbonylindole Derivatives.
A p-toluenesulfonic acid-catalyzed cascade reaction is reported for the synthesis of 4-functionalized tetrahydrocarbazolones via the reaction of 4-(indol-2-yl)-4-oxobutanal derivatives with a variety of nucleophiles in acetonitrile or hexafluoroisopropanol. After the initial intramolecular Friedel-Crafts hydroxyalkylation, the 3-indolylmethanol intermediate is subsequently activated and reacted with the external nucleophile. The reaction conditions are crucial to avoid alternative reaction pathways, allowing direct substitution reaction with thiols, (hetero)arenes, alkenes, or sulfinates. The procedure features high overall yields to access a diverse family of compounds bearing the tetrahydrocarbazole core.


Vibrio vulnificus and Vibrio parahaemolyticus in Oysters under Low Tidal Range Conditions: Is Seawater Analysis Useful for Risk Assessment?
Human-pathogenic Vibrio bacteria are acquired by oysters through filtering seawater, however, the relationships between levels of these bacteria in measured in oysters and overlying waters are inconsistent across regions. The reasons for these discrepancies are unclear hindering our ability to assess if -or when- seawater samples can be used as a proxy for oysters to assess risk. We investigated whether concentrations of total and human pathogenic Vibrio vulnificus (vvhA and pilF genes) and Vibrio parahaemolyticus (tlh, tdh and trh genes) measured in seawater reflect concentrations of these bacteria in oysters (Crassostrea virginica) cultured within the US lower Chesapeake Bay region. We measured Vibrio spp. concentrations using an MPN-qPCR approach and analyzed the data using structural equation modeling (SEM). We found seawater concentrations of these bacteria to predictably respond to temperature and salinity over chlorophyll a, pheophytin or turbidity. We also inferred from the SEM results that Vibrio concentrations in seawater strongly predict their respective concentrations in oysters. We hypothesize that such seawater-oyster coupling can be observed in regions of low tidal range. Due to the ease of sampling and processing of seawater samples compared to oyster samples, we suggest that under low tidal range conditions, seawater samples can foster increased spatial and temporal coverage and complement data associated with oyster samples.


Scattering and interference in epitaxial graphene.
A single sheet of carbon, graphene, exhibits unexpected electronic properties that arise from quantum state symmetries, which restrict the scattering of its charge carriers. Understanding the role of defects in the transport properties of graphene is central to realizing future electronics based on carbon. Scanning tunneling spectroscopy was used to measure quasiparticle interference patterns in epitaxial graphene grown on SiC(0001). Energy-resolved maps of the local density of states reveal modulations on two different length scales, reflecting both intravalley and intervalley scattering. Although such scattering in graphene can be suppressed because of the symmetries of the Dirac quasiparticles, we show that, when its source is atomic-scale lattice defects, wave functions of different symmetries can mix.


Central Corneal Thickness and Intraocular Pressure in Children with Type 1 Diabetes Mellitus.
The aim of this study is to determine the mean central corneal thickness (CCT) and mean intraocular pressure (IOP) in children with type 1 diabetes mellitus (T1DM) and to determine the relationship between CCT and IOP on the one hand and age, sex, retinopathy hemoglobin A1c (HbA1c), and duration of diabetes on the other. This is a case-control, hospital-based study conducted at Hospital Universiti Sains Malaysia between January and November 2022. Thirty-eight children with T1DM were recruited as cases, and 38 healthy children were recruited as controls. The cases and controls then underwent ophthalmic examination, IOP measurement, and CCT measurement using optical coherence tomography (OCT) of the right eye. The IOP measurements were adjusted for CCT for further analysis. The means of CCT and IOP values were significantly higher in the T1DM group than in the control group (all p = 0.02). The mean CCT was 542.18 ± 20.40 μm in the T1DM group, and 529.52 ± 26.17 μm in the control group. The mean IOP was 14.68 ± 1.98 mmHg in the T1DM group, and 13.52 ± 1.66 mmHg in the control group. The mean HbA1c was 10.68% ± 2.49% in the T1DM group. Age and duration of DM were found to have a significant association with CCT in children with T1DM. The duration of DM was also found to be significantly associated with the IOP. Sex and HbA1c levels were found to have no significant relationship with either CCT or IOP. Children with T1DM have significantly higher CCT and IOP than the average child. The duration of DM is a significant factor that impacts both CCT and IOP. In addition, age is another factor that affects CCT in children with T1DM.


Reconceptualizing Stigmas in Aged Care: A Typology of Stigmatizing Discourses in the Aged-Care Context.
Stigmatizing discourses surrounding aged care is complex. The stigma associated with people working in aged care is often conflated with negative evaluations about older adults who receive care and/or the institutions and systems that provide these services. This lack of nuance hampers productive dialogue in gerontology required for making sense of the stigmas attached to entities and systems. We propose that an interdisciplinary approach drawing on gerontology, psychology, and linguistics would facilitate a greater understanding of the stigma of aged care. Specifically, a discourse approach underpinned by Stigma Theory and Systemic Functional Linguistics can clarify the different stigmatizing discourses associated with aged-care entities and systems, and inform priorities for interventions targeting each discourse category identified. APPRAISAL, a linguistics framework, was employed to analyze evaluative language in 660 submissions published in the Australian Royal Commission into Aged Care. Through applying predefined language-based criteria, we distinguished between multiple stigmatizing discourses that were interrelated but conceptually distinct in terms of attitude types and targets. Twelve categories of stigmatizing discourses derived from the linguistic analysis established a typology. This typology conceptualized different ways stigma can be meaningfully understood in the aged-care context. By employing this typology, 12 intervention priorities were also identified. This interdisciplinary approach enabled us to examine the diversity and complexity of the social construction of stigma in the public domain. We argue that understanding these linguistic patterns can assist with designing targeted aged-care interventions and policies.


The role of interphotoreceptor retinoid-binding protein on the translocation of visual retinoids and function of cone photoreceptors.
The first event in light perception is absorption of a photon by the retinaldehyde chromophore of an opsin pigment in a rod or cone photoreceptor cell. This induces isomerization of the chromophore, rendering the bleached pigment insensitive to light. Restoration of light sensitivity requires chemical reisomerization of retinaldehyde via a multistep enzyme pathway, called the visual cycle, in cells of the retinal pigment epithelium (RPE). Interphotoreceptor retinoid-binding protein (IRBP) is present in the extracellular space between photoreceptors and the RPE. IRBP is known to bind visual retinoids. Previous studies on irbp(-/-) mice suggested that IRBP plays an insignificant role in opsin-pigment regeneration. However, the mice in these studies were uncontrolled for a severe mutation in the rpe65 gene. Rpe65 catalyzes the rate-limiting step in the visual cycle. Here, we examined the phenotype in irbp(-/-) mice homozygous for the wild-type (Leu450) rpe65 gene. We show that lack of IRBP causes delayed transfer of newly synthesized chromophore from RPE to photoreceptors. Removal of bleached chromophore from photoreceptors is also delayed in irbp(-/-) retinas after light exposure. It was previously shown that rods degenerate in irbp(-/-) mice. Here, we show that cones and rods degenerate at similar rates. However, cones are more affected functionally and show greater reductions in outer segment length than rods in irbp(-/-) mice. The disproportionate reductions in cone function and outer-segment length appear to result from mistrafficking of cone opsins due to impaired delivery of retinaldehyde chromophore, which functions as a chaperone for cone opsins but not rhodopsin.


Age differences in the effects of conceptual integration training on resource allocation in sentence processing.
This research examined age differences in the accommodation of reading strategies as a consequence of explicit instruction in conceptual integration. In Experiment 1, young, middle-aged, and older adults read sentences for delayed recall using a moving-window method. Readers in an experimental group received instruction in making conceptual links during reading while readers in a control group were simply encouraged to allocate effort. Regression analysis to decompose word-by-word reading times in each condition isolated the time allocated to conceptual processing at the point in the text at which new concepts were introduced, as well as at clause and sentence boundaries. While younger adults responded to instructions by differentially allocating effort to sentence wrap-up, older adults allocated effort to intrasentence wrap-up and on new concepts as they were introduced, suggesting that older readers optimized their allocation of effort to linguistic computations for textbase construction within their processing capacity. Experiment 2 verified that conceptual integration training improved immediate recall among older readers as a consequence of engendering allocation to conceptual processing.


Decision-making in the physician-patient encounter: revisiting the shared treatment decision-making model.
In this paper we revisit and add elements to our earlier conceptual framework on shared treatment decision-making within the context of different decision-making approaches in the medical encounter (Charles, C., Gafni, A., Whelan, T., 1997. Shared decision-making in the medical encounter: what does it mean? (or, it takes at least two to tango). Social Science & Medicine 44, 681 692.). This revised framework (1) explicitly identifies different analytic steps in the treatment decision-making process; (2) provides a dynamic view of treatment decision-making by recognizing that the approach adopted at the outset of a medical encounter may change as the interaction evolves; (3) identifies decision-making approaches which lie between the three predominant models (paternalistic, shared and informed) and (4) has practical applications for clinical practice, research and medical education. Rather than advocating a particular approach, we emphasize the importance of flexibility in the way that physicians structure the decision-making process so that individual differences in patient preferences can be respected.


Gambling behavior among Macau college and university students.
This survey investigated gambling behavior among Chinese students studying in Macau colleges and universities. It also aimed to examine the relationship between problem gambling, affect states and sensation seeking propensity. A convenience sample of 999 students (370 men, 629 women) filled a self-administered questionnaire consisted of the Problem Gambling Severity Index (PGSI) (Ferris and Wynne in The Canadian problem gambling index: User manual. Canadian Centre on Substance Abuse, Toronto 2001a), the 8-item Brief Sensation Seeking Scale (BSSS-8) (Hoyle et al. Pers Individ Diff 32(3): 401-414, 2002), Bradburn's Affect Balance Scale (BABS) (Bradburn in The structure of psychological well-being. Aldine, Chicago 1969) and questions on gambling activities. The response rate is 65%. Results indicate 32.3% (n = 323) of the survey participants wagered on mahjong (61.8%), soccer matches (40.2%), Mark Six lottery (37.2%), card games (28.1%), land-based casino gambling (13.1%), slot machines (7.5%) and online casino games (2.0%). The average monthly stake was MOP $411. Seeking entertainment (18.7%), killing time (12.5%) and peer influence (11.1%) were the three main reasons for gambling. Using the PGSI, 3.6 and 5.3% of the students could be identified as moderate-risk and problem gamblers respectively. Men were significantly more vulnerable to gambling problems (X2(1) = 35.00, p < 0.01) than women. Most of the problematic gamblers (76%) made their first bet before 14 years. The PGSI scores are significantly correlated with the BSSS-8 scores (r = 0.23, p < 0.01) but not with the overall ABS scores (r = -0.06, p > 0.05). The study findings inform campus prevention programs and future research.


Modified antigens in the treatment of allergic disease.
In summary, effects at improvement upon conventional immunotherapy by antigen modification have taken three major directions. First, methods designed primarily to impede allergen release from the site of deposition have been only modestly successful. An example is alum-precipitated extracts which require somewhat fewer injections. Second, attempts to suppress specific IgE production have been unsuccessful in human trials, although antigen-specific IgG levels have increased in a manner similar to that produced by conventional allergen immunotherapy. The third major avenue of antigen modification, to reduce allergenicity while retaining immunogenicity, appears, at present, to be the most promising. Clinical studies of polymerized allergens summarized above have shown that patients can be given a course of polymerized allergen immunotherapy in a few weeks, while a course involving a comparable quantity of unmodified allergen would require years. With the use of polymerized pollen extracts, degree of symptom reduction, persistence of benefit, and magnitude of IgG-blocking antibody induction are comparable to that achieved with unmodified allergens, while the incidence of systemic reactions is greatly reduced. The development of polymerized allergens appears to represent a significant advance which may be expected to expand the clinical utility of allergen immunotherapy.


In silico identification of potential SARS COV-2 2'-O-methyltransferase inhibitor: fragment-based screening approach and MM-PBSA calculations.
In the present era, there are many efforts trying to face the emerging and successive waves of the COVID-19 pandemic. This has led to considering new and unusual targets for SARS CoV-2. 2'-O-Methyltransferase (nsp16) is a key and attractive target in the SARS CoV-2 life cycle since it is responsible for the viral RNA protection via a cap formation process. In this study, we propose a new potential inhibitor for SARS COV-2 2'-O-methyltransferase (nsp16). A fragment library was screened against the co-crystal structure of the SARS COV-2 2'-O-methyltransferase complexed with Sinefungin (nsp16 - PDB ID: 6WKQ), and consequently the best proposed fragments were linked via a de novo approach to build molecule AP-20. Molecule AP-20 displayed a superior docking score to Sinefungin and reproduced the key interactions in the binding site of 2'-O-methyltransferase. Three molecular dynamic simulations of the 2'-O-methyltransferase apo structure and its complexed forms with AP-20 and Sinefungin were performed for 150 nano-seconds to provide insights on the dynamic nature of such setups and to assess the stability of the proposed AP-20/enzyme complex. AP-20/enzyme complex demonstrated better stability for the ligand-enzyme complex compared to Sinefungin in a respective setup. Furthermore, MM-PBSA binding free energy calculations showed a better profile for AP-20/enzyme complex compared to Sinefungin/enzyme complex emphasizing the potential inhibitory effect of AP-20 on SARS COV-2 2'-O-methyltransferase. We endorse our designed molecule AP-20 to be further explored via experimental evaluations to confront the spread of the emerging COVID-19. Also, in silico ADME profiling has ascribed to AP-20 an excellent safety and metabolic stability profile.


Causality Assessment Guidelines for Adverse Events Following Immunization with a Focus on Guillain-Barré Syndrome.
South Korea operates a National Vaccine Injury Compensation Program (VICP) for people who experience adverse events following immunization (AEFI). To run this program rationally, it is a prerequisite to confirm whether adverse events were caused by immunization. Guillain-Barré syndrome (GBS), a severe neurological disease with limb pain and muscle weakness as cardinal symptoms, is attracting attention as an AEFI. However, algorithm or guidelines for assessing the causality between vaccination and the incidence of GBS are lacking. We aimed to develop guidelines for causality assessment of GBS as an AEFI and suggest using these guidelines in alignment with the VICP. We systematically searched for other previously published algorithms or guidelines and found a WHO-AEFI guideline used worldwide; however, it only provides general instructions and is not tailored to specific adverse events. We translated and locally adapted the structure of this guideline and then added contents related to GBS. The GBS-specific guideline consists of four steps: case ascertainment of GBS, checklist (including (1) order of incidence, (2) temporal proximity, (3) evidence for other causes and (4) published evidence), an algorithm, and final classification. We listed key information on confirming GBS and whether any other causes of GBS were present. For real world application of the guideline along with the VICP, we collaborated with a panel of neurologists, epidemiologic investigators, and committee members from the VICP. To ensure transparency and a scientific approach, regular updates and collaboration with neurologists are essential. We expect that this guideline will contribute to logical causality assessment and compensation decisions for GBS and will provide the basic structure for causality assessment of other AEFIs.


Isolation and identification of Acanthamoeba genotypes and Naegleria spp. from the water samples of public swimming pools in Qazvin, Iran.
Free-living amoeba (FLA), including Acanthamoeba and Naegleria are facultative parasites in humans. The amoeba have widespread distribution in various water sources. The aim of this study was isolation and molecular identification of Acanthamoeba and Naegleria isolated from swimming pools and also hot and cold tub waters in Qazvin province. The samples (166 water samples) were cultured to isolate and identify positive specimens. PCR (polymerase chain reaction) amplification, sequencing and phylogenetic analysis were conducted to confirm the isolated species and genotypes of amoeba. According to morphological characterizations, 18.6% of specimens were identified as FLA, which in 71% were Acanthamoeba by PCR method. Molecular analysis revealed that 36.3%, 18.1% and 4.5% of Acanthamoeba specimens were identified as T3, T4 and T11 Acanthamoeba genotypes, respectively. Protacanthamoeba bohemica (27.2%) and Acanthamoeba sp. (4.5%) were found among the specimens. The results of osmo-tolerance and thermo-tolerance assays demonstrated that 50% of T3 and 25% of T4 genotypes of Acanthamoeba were highly pathogenic parasites. The molecular approach showed the presence of Naegleria lovaniensis (9%) in hot tub water of swimming pools. This study demonstrated that the swimming pools and hot tub water in Qazvin province were contaminated with Acanthamoeba and Naegleria species.


[Evaluation of antiepileptic effects of cortexin in a model of convulsions].
We studied antiepileptic effects of cortexin administered in doses 0,015, 0,15 and 1,0 mg/kg intraperitoneally in solution or intranasally in the complex with nanoparticles in a model of acute and chronic convulsions in rats induced by pentylenetetrazole. In the model of epileptic status, the long-term preliminary administration of cortexin had no effect on convulsions while in the model of chronic convulsions (temporal epilepsy), cortexin had a marked dose-dependent antiepileptic effect. The influence of cortexin on neuroplasticity and its clinical potential are discussed.


[Utilization of technological resources within the framework of operation of a Mobile Mental Health Unit].
Telepsychiatry was introduced in the early 1950's for the provision of mental health services from a distance. In 1990 the progress made in telecommunications technologies caused a significant expansion in telepsychiatry services. It can refer to store and forward technologies, interactive technologies, remote monitoring technologies and it is applied to contribute to the lift of restrictions placed on providing mental health services. Restrictions may exist due to geographic isolation, lack of specialized services, high cost of moving patients etc. The positive cost-benefit analysis and the reliability of diagnosis and efficacy of interventions through telepsychiatry have been documented in various research papers referring to a wide range of contexts such as prisons, remote areas, general and psychiatric hospitals. Since 2003 the Mobile Mental Health Unit of South- Eastern (SE) Cyclades has been using videoconferencing in order to provide mental health services in thirteen islands. This area shares many of the characteristics of remote areas such as residential dispersion, lack of mental health services and frequent lack of access to services in urban centers. Telepsychiatry in conjunction with physical presence of professionals was launched by the Mobile Mental Health Unit in order to provide assistance to the evaluation of patients, to therapeutic interventions, to medicine prescription, to crisis intervention, to psychoeducation of patients and their families and to the implementation of educational and administrative activities . The use of existing technology in combination with the development of human resources has enabled continuity of care, crisis intervention and avoidance of involuntary hospitalization for a significant number of persons. Moreover, it has improved cooperation and coordination between the interdisciplinary team and local authorities and agencies. When professionals located in different parts are collaborating via telepsychiatry it is necessary to ensure that they have immediate and simultaneous access to data concerning the patient. This need is served by the electronic medical record that facilitates access to patient records, improves the quality of care, reduces the probability of making wrong decisions and contributes to the promotion of research and service evaluation. KLIMAKA's electronic medical record contains more than ten thousand cases of which 4544 are related to individuals served by the Mobile Mental Health Unit during the years 2003-2010. This article presents the basic functions and parameters of the electronic medical record, the applications of telepsychiatry in the whole range of services provided by the scientific staff and provides information on how human and technology resources are distributed to the branches of the Mobile Mental Health Unit of SE Cyclades.


2-Methyl-4-chlorophenoxyacetic acid and bromoxynil herbicide death.
We report a fatal case of a 37 year old gentleman who ingested a MCPA/bromoxynil co-formulation herbicide. Although clinically well on initial examination, our patient declined dramatically over his 18 h admission with increasing CO2 production, hyperthermia and metabolic derangement to eventually die from cardiac asystole 20 h post ingestion. Two hours after ingestion the MCPA concentration was 83.9 μg/mL and bromoxynil concentration was 137 μg/mL. The patients' mechanism of death appeared to be uncoupling of oxidative phosphorylation, excess CO2 production and hyperthermia. There is limited knowledge on the acute toxicity of these herbicides, in particular bromoxynil, and this case highlights the relentless progression of severe toxicity in humans.


Short-term interventions on wards fail to reduce preanalytical errors: results of two prospective controlled trials.
Preventing laboratory errors promotes patient safety and reduces the cost of unnecessary processing. The aim of this study was to test the effectiveness of two short-term interventions at reducing errors in the preanalytical stage of laboratory testing. Error data were reviewed from inpatient wards at Bradford Royal Infirmary (BRI), Leeds General Infirmary (LGI) and St James' University Hospital (SJUH) for 22 weeks. Two separate interventions lasted for two weeks. The outcome measures were inadequate tube and form labelling, incorrect tube selection and insufficient sample volume. Posters targeting these errors were created and displayed on inpatient wards in SJUH (n = 48). BRI and LGI were control hospitals. Qualitative interviews were held with clinical staff to raise awareness of common errors, give advice and discuss error reduction (n = 37). Ten weeks later, screensavers warning against labelling errors were displayed (LGI and SJUH). Quantitative error data, routinely collected by the laboratory, were used for analysis. There was no change in error rate or type at the intervention site(s) compared with the control(s). There were 7058 reported errors across three sites, of which 6623 were errors targeted by the interventions. The overall error rate remained stable on all three sites (analysis of variance, P = 1.0). When interviewing clinical staff, 29% thought that equipment was the main contributing factor to errors while 23% struggled with tube selection. Despite enthusiasm on the part of the ward-based staff, both short-term interventions had no significant impact on preanalytical error rates. Most errors are due to human factors. These may be reduced with the introduction of an electronic ordering system.