Changes in the distribution of microtubules and intermediate filaments in mammalian Sertoli cells during spermatogenesis.
We have studied the distribution of microtubules and intermediate filaments in mammalian Sertoli cells during spermatogenesis. The arrangement of microtubules was determined, by indirect immunofluorescence, in ground squirrel testes that were 1) fixed, mechanically fragmented, and attached to polylysine-coated slides, and 2) fixed, embedded in polyethylene glycol, and sectioned. Intermediate filament patterns were determined, also by indirect immunofluorescence, in sections of unfixed rat testis. Results from these studies were confirmed and extended using electron microscopy. Microtubules first become evident in lateral processes that embrace round spermatids. When spermatids elongate and become situated in apical crypts of Sertoli cells, the microtubules become oriented parallel to the long axis of Sertoli cells and surround the crypts. As spermatids mature and acquire a saucer shape, apical microtubules progressively concentrate in Sertoli cell regions adjacent to the acrosome and eventually form discrete C-shaped structures that disappear during spermiation. Intermediate filaments in rat Sertoli cells are centered around the nucleus. From perinuclear regions, filaments extend toward desmosome-like junctions with early spermatogenic cells and into the apical cytoplasm where they have a transient association with crypts containing elongate spermatids. Filaments amongst crypts are most evident in early stages of the spermatogenic cycle when apical crypts are situated deep within the epithelium. They become less evident and eventually disappear as spermatids assume a more apical position. Our fluorescence studies and ultrastructural analyses indicate that the association of intermediate filaments with crypts is specific to regions adjacent to the dorsal or convex aspect of spermatid heads. In these regions, approximately 8 to 12 uniformly aligned filaments are intimately associated with actin filaments in ectoplasmic specializations surrounding the crypts. We conclude that, like actin, the distribution of microtubules and intermediate filaments changes in Sertoli cells during spermatogenesis. The distribution of microtubules correlates with the irregular columnar shape of Sertoli cells. We suspect that the apically situated intermediate filaments may play a role in anchoring or positioning Sertoli cell crypts deep within the epithelium during the early stages of the spermatogenic cycle.


7α-Hydroxypregnenolone regulates diurnal changes in sexual behavior of male quail.
In the Japanese quail, 7α-hydroxypregnenolone, a previously undescribed avian neurosteroid, is actively produced in the brain. 7α-Hydroxypregnenolone acts as a novel neuronal activator to stimulate locomotor activity of quail. Therefore, in this study, we determined whether 7α-hydroxypregnenolone changes the expression of sexual behavior in Japanese quail. We first measured diurnal changes in sexual behavior of male quail exposed to a long-day photoperiod. We found that sexual behavior of male quail was high in the morning when endogenous 7α-hydroxypregnenolone level is high. Subsequently, we centrally administered 7α-hydroxypregnenolone in the evening when endogenous 7α-hydroxypregnenolone level is low. In the 30 min after intracerebroventricular (ICV) injection, 7α-hydroxypregnenolone dose dependently increased the frequency of sexual behavior of male quail. However, 7β-hydroxypregnenolone, a stereoisomer of 7α-hydroxypregnenolone, did not effect on the frequency of sexual behavior of male quail. In addition, to confirm the action of 7α-hydroxypregnenolone on sexual behavior, male birds received an ICV injection of ketoconazole, an inhibitor of cytochrome P450s, and behavioral experiments were performed in the morning. Ketoconazole significantly decreased the frequency of sexual behavior of male quail, whereas administration of 7α-hydroxypregnenolone to ketoconazole-treated males increased the frequency of their sexual behavior. These results indicate that 7α-hydroxypregnenolone regulates diurnal changes in sexual behavior of male quail.


In vivo and in vitro apoptosis induced by new acaricidal ethyl-carbamates in Rhipicephalus microplus.
The ability of ethyl-4-bromophenylcarbamate (LQM 919) and ethyl-4-chlorophenylcarbamate (LQM 996) to induce in vivo apoptosis of Rhipicephalus microplus ovarian cells and in vitro apoptosis of tick and mammalian cell culture was evaluated. The ovaries of engorged females treated with 1 mg mL-1 LQM 919 or LQM 996 presented more (p < 0.001) peroxidase-TUNEL-positive labeled cells (apoptotic cells) in situ than their respective control groups, and this increase was time-dependent (p < 0.001). The majority of apoptotic cells were observed in the epithelium and ovarian pedicel. HepG2, Vero and Rm-sus cells, as well as cells from primary cultures of R. microplus salivary glands, intestine and ovaries were exposed to different concentrations of the ethyl-carbamates. Both ethyl-carbamates induced a concentration-dependent reduction in the viability of all cell types (p < 0.001). Exposure to the ethyl-carbamates increased caspase 3 activity (p < 0.01) in primary cultures and cell lines, except in HepG2 cells. Fluorescent TUNEL-positive cells were observed in all cell types treated with 600 μM LQM 919 or LQM 996. These results indicate that both ethyl-carbamates induce apoptosis of the ovarian, intestinal and salivary glands cells in R. microplus and strongly suggest that this is their main mechanism of acaricidal action.


Coexisting Coronary and Carotid Artery Disease - Which Technique and in Which Order? Case Report and Review of Literature.
Coexisting carotid artery stenosis and coronary artery disease is common and there is currently no consensus in treatment guidelines on the timing, sequence and methods of revascularization. We report a case of a patient with symptomatic triple vessel coronary artery disease as well as asymptomatic severe right internal carotid artery stenosis. Our patient underwent myocardial revascularization first, because she presented with unstable angina and was asymptomatic neurologically. This article summarizes current literature about the approach to carotid and coronary artery revascularization and addresses the decision-making process regarding the timing and sequence of revascularization.


Recurrent submacular hemorrhage in patients with neovascular age-related macular degeneration.
To describe the incidence, risk factors for, and long-term visual outcomes of recurrent submacular hemorrhage in the context of age-related macular degeneration. Medical records of patients with neovascular age-related macular degeneration with or without polypoidal choroidal vasculopathy showing submacular hemorrhage at their first visit to our institution were reviewed. The required minimum follow-up period was 24 months, and any newly developed submacular hemorrhage larger than 1 disk area after near-complete resolution of initial hemorrhage was defined as recurrence. A total of 47 eyes of 47 patients were eligible for inclusion. Twenty-four patients showed recurrent submacular hemorrhage during the follow-up period (Group I). Patients without recurrent submacular hemorrhage were included in Group II. The time to recurrent submacular hemorrhage in Group I patients was 21.4 ± 9.2 months. Polypoidal choroidal vasculopathy was present in 50% of Group I patients (n = 12) and 13% of Group II patients (n = 3) (P = 0.025). Intravitreal anti-vascular endothelial growth factor injection was performed during the follow-up period in 70.8% of Group I patients (n = 17) and 95.7% of Group II patients (n = 22) (P = 0.048). Visual acuity change during the follow-up period did not significantly differ between the two groups. In patients with neovascular age-related macular degeneration presenting with submacular hemorrhage at their first visit, the incidence of recurrent submacular hemorrhage was 51.1% in our retrospective long-term follow-up study. The presence of polypoidal choroidal vasculopathy was associated with an increased risk of recurrent submacular hemorrhage. Use of anti-vascular endothelial growth factor agents was correlated with a reduced risk of such hemorrhage. Visual acuity was stably maintained over 2 years regardless of hemorrhage recurrence.


Alteration in risk factor accumulations of acute myocardial infarction during the last one decade: analysis of patients admitted in Coronary Care Unit.
The present study was undertaken to determine accumulation of risk factors in acute myocardial infarction during two periods of 2002 and 1990-1991. We collected 173 and 153 patients with acute myocardial infarction in 2002 and 1990-1991, respectively, and analyzed the history of multiple risk factors, including diabetes mellitus, impaired glucose tolerance, hyperlipidemia, hypertension and obesity, and laboratory findings. The numbers and their percentages of all the risk factors increased in 2002 compared with 1990-1991. According to plasma glucose level, the patients who had type 2 diabetes mellitus, and impaired fasting glucose or impaired glucose tolerance had increased markedly from 41 to 65%. Multiple accumulation of risk factors had increased during the last one decade, and only one or no risk factor per se was not the case in the patients with acute myocardial infarction. Hyperlipidemia and hypertension became fairly controlled in the patients, but not hyperglycemia in type 2 diabetes mellitus in the period of 2002. These findings may indicate that increased multiple accumulation of risk factors accelerates the occurrence of acute myocardial infarction in 2002 as compared to 1990-1991.


Semiquantitative assessment of subchondral bone marrow edema-like lesions and subchondral cysts of the knee at 3T MRI: a comparison between intermediate-weighted fat-suppressed spin echo and Dual Echo Steady State sequences.
Choice of appropriate MR pulse sequence is important for any research studies using imaging-derived data. The aim of this study was to compare semiquantitative assessment of subchondral bone marrow edema-like lesions and subchondral cysts using intermediate-weighted (IW) fat-suppressed (fs) spin echo and Dual Echo Steady State (DESS) sequences on 3 T MRI. Included were 201 subjects aged 35-65 with frequent knee pain. 3T MRI was performed with the same sequence protocol as in the Osteoarthritis Initiative (OAI). In a primary reading subchondral bone marrow edema-like lesions were assessed according to the WORMS system. Two hundred subregions with such lesions were randomly chosen. The extent of subchondral bone marrow edema-like lesions was re-evaluated separately using sagittal IW fs and DESS sequences according to WORMS. Lesion size and confidence of the differentiation between subchondral bone marrow edema-like lesions and subchondral cysts located within or adjacent to them was rated from 0 to 3. Wilcoxon signed-rank tests and chi-square statistics were used to examine differences between the two sequences. Of 200 subchondral bone marrow edema-like lesions detected by IW fs sequence, 93 lesions (46.5%) were not depicted by the DESS sequence. The IW fs sequence depicted subchondral bone marrow edema-like lesions to a larger extent than DESS (p < 0.0001), and the opposite was true for subchondral cysts. Confidence scores for differentiation of the two types of lesions were not significantly different between the two sequences. In direct comparison the IW fs sequence depicts more subchondral bone marrow edema-like lesions and better demonstrate the extent of their maximum size. The DESS sequence helps in the differentiation of subchondral bone marrow edema-like lesions and subchondral cysts. The IW fs sequence should be used for determination of lesion extent whenever the size of subchondral bone marrow edema-like lesions is the focus of attention.


[Implementation of a specific premedication protocol for tracheal intubation in the delivery room. Practice in two level-III hospitals].
Tracheal intubation in neonates is a painful procedure performed daily in the delivery room despite the widespread development of noninvasive ventilation. Specific analgesia is not commonly performed. The objective of this observational study was to compare practices between two level-III centers: one with a specific protocol for premedication before tracheal intubation of newborns in the delivery room, the other without. One hundred and fifteen neonates were intubated in the delivery room and included over a 4-month period: 25% of them received specific premedication before intubation, exclusively in the center with the protocol. None of the extreme premature neonates (age≤28 gestational weeks) received analgosedation before the procedure. Nalbuphine, midazolam, and sufentanil were mainly used, via the intravenous or intrarectal route. Infants receiving a premedication were significantly heavier and had a greater gestational age than the others (1500 g [range, 1180-2260 g] vs. 1170 [range, 860-1680 g] P=0.003, and 31 GW [range, 29-34 GW] vs. 29 [range, 27-32 GW] P=0.014, respectively). Most pediatricians (85-100%) favored a specific protocol for sedation before tracheal intubation. Implementation of a specific protocol allows specific analgesia to be implemented for newborns undergoing tracheal intubation. Further studies should be conducted to determine the best strategies for pain management during tracheal intubation of neonates, especially in the delivery room.


Circulating Unsaturated Fatty Acids Delineate the Metabolic Status of Obese Individuals.
Obesity is not a homogeneous condition across individuals since about 25-40% of obese individuals can maintain healthy status with no apparent signs of metabolic complications. The simple anthropometric measure of body mass index does not always reflect the biological effects of excessive body fat on health, thus additional molecular characterizations of obese phenotypes are needed to assess the risk of developing subsequent metabolic conditions at an individual level. To better understand the associations of free fatty acids (FFAs) with metabolic phenotypes of obesity, we applied a targeted metabolomics approach to measure 40 serum FFAs from 452 individuals who participated in four independent studies, using an ultra-performance liquid chromatograph coupled to a Xevo G2 quadruple time-of-flight mass spectrometer. FFA levels were significantly elevated in overweight/obese subjects with diabetes compared to their healthy counterparts. We identified a group of unsaturated fatty acids (UFAs) that are closely correlated with metabolic status in two groups of obese individuals who underwent weight loss intervention and can predict the recurrence of diabetes at two years after metabolic surgery. Two UFAs, dihomo-gamma-linolenic acid and palmitoleic acid, were also able to predict the future development of metabolic syndrome (MS) in a group of obese subjects. These findings underscore the potential role of UFAs in the MS pathogenesis and also as important markers in predicting the risk of developing diabetes in obese individuals or diabetes remission after a metabolic surgery.


In vitro studies of hypoxia inducible factor-prolyl hydroxylase inhibitors daprodustat, desidustat, and vadadustat for equine doping control.
Performance-enhancing substances and methods have become a serious problem in competitive sports. The hypoxia-inducible factor (HIF) stabilizers can enhance the organism's capacity for molecular oxygen transport and are likely to be abused as performance-enhancing agents in sports. This paper describes the metabolic conversion of the popular hypoxia inducible factor-prolyl hydroxylase (HIF-PH) inhibitors, namely, daprodustat, desidustat, and vadadustat using equine liver microsomes, determined on a QExactive high-resolution mass spectrometer. During this study, a total of 10 metabolites for daprodustat (all are Phase I), 10 metabolites for desidustat (five each for Phase I and Phase II), and 15 metabolites for vadadustat (six for Phase I and nine for Phase II) were detected. The important findings of the current research are as follows: (1) All the three HIF-PH inhibitor drug candidates are prone to oxidation, which results in corresponding hydroxylated metabolites; (2) in desidustat, hydrolysis and dissociation of oxime linkage also observed; (3) the glucuronic acid conjugate (except daprodustat) of the parent drugs as well as the monohydroxylated analogs were observed; (4) sulfonic acid conjugated metabolites were observed only for vadadustat.


Definition by FSH, AMH and embryo numbers of good-, intermediate- and poor-prognosis patients suggests previously unknown IVF outcome-determining factor associated with AMH.
Though outcome models have been proposed previously, it is unknown whether cutoffs in clinical pregnancy and live birth rates at all ages are able to classify in vitro fertilization (IVF) patients into good-, intermediate- and poor prognosis. We here in 3 infertile patient cohorts, involving 1247, 1514 and 632 women, built logistic regression models based on 3 functional ovarian reserve (FOR) parameters, including (1) number of good quality embryos, (2) follicle stimulating hormone (FSH, mIU/mL) and (3) anti-Müllerian hormone (AMH, ng/mL), determining whether clinical pregnancy and live birth rates can discriminate between good, intermediate and poor prognosis patients. All models, indeed, allowed at all ages for separation by prognosis, though cut offs changed with age. In the embryo model, increasing embryo production resulted in linear improvement of IVF outcomes despite transfer of similar embryo numbers; in the FSH model outcomes and FSH levels related inversely, while the association of AMH followed a bell-shaped polynomial pattern, demonstrating "best" outcomes at mid-ranges. All 3 models demonstrated increasingly poor outcomes with advancing ages, though "best" AMH even above age 43 was still associated with unexpectedly good pregnancy and delivery outcomes. Excessively high AMH, in contrast, was at all ages associated with spiking miscarriage rates. At varying peripheral serum concentrations, AMH, thus, demonstrates hithero unknown and contradictory effects on IVF outcomes, deserving at different concentrations investigation as a potential therapeutic agent, with pregnancy-supporting and pregnancy-interrupting properties.


Cloning and expression of a common Legionella outer membrane antigen in Escherichia coli.
A genomic library of Legionella pneumophila was constructed by inserting L. pneumophila knoxville-1 strain (LPK-1) chromosome fragments into cosmid vector pHC79. Screening of the library with antibodies directed against a major outer membrane protein/lipopolysaccharide complex from LPK-1 resulted in the identification of six clones that reacted with the antiserum. Western blot analysis indicated that a 19,000 dalton (19 kDa) component was the reactive antigen in all of the clones. Western blot analysis of outer membranes from L. pneumophila serogroups and other Legionella species revealed that the cloned 19 kDa antigen was common to all serogroups and all but one of the five other Legionella species examined. One of the 19 kDa expressing clones was used as an immunoabsorbent to recover antibody to the 19 kDa antigen thus confirming the surface localization of this L. pneumophila antigen in E. coli.


Treating late-life depression: Comparing the effects of internet-delivered cognitive behavior therapy across the adult lifespan.
The burden attributable to late-life depression is set to increase with the ageing population. The size of the workforce trained to deliver psychogeriatric medicine is limited. Internet-delivered cognitive behavioral therapy (iCBT) is an efficacious and scalable treatment option for depression. Yet older adults are underrepresented in iCBT research. This study examines the effects of iCBT for depression across the adult lifespan among patients seeking help in routine clinical care (N = 1288). Regression analyses were used to examine the relationship between age group (e.g., 18-24years (n = 141); 25-34years (n = 289); 35-44years (n = 320); 45-54years (n = 289); 55-64years (n = 180); 65 +years (n = 69)) and presenting demographic and clinical characteristics, adherence to treatment, and rates of remission, recovery and reliable improvement. Linear mixed models were used to examine whether reductions in symptom severity, distress and impairment varied as a function of age. Patients aged 65+ years were more likely to be male compared to those aged 18-34 years and have been prescribed iCBT by their GP compared to those aged 55-64 years. Patients experiencing late-life depression experienced moderate to large effect size reductions in depressive symptom severity, psychological distress, and impairment, as did all other age groups. Rates of remission, recovery or reliable improvement were comparable across the adult lifespan. iCBT is an effective treatment option for depression including in later life, and can be used to scale evidenced-based medicine in routine clinical care. No follow-up data were collected. The long-term effects of treatment, particularly for those who did not experience remission, are unclear.


A near-infrared Fluorescent Probe Based on Dicyanisophorone for the Detection of Zinc Ions (Zn2+) in Water and Living Cells.
As one of the important metal ions, zinc ions (Zn2+) are widely involved in various physiological and pathological processes, and play fundamental roles in neurotransmission, cell metabolism and apoptosis. However, the convenient monitor of Zn2+ in environmental and biological samples remains challenging. In this study, a small molecule dicyanoisophorone-based schiff base incorporating with o-phenylenediamine was synthesized. It can rapidly combine with Zn2+ to emit significant near-infrared fluorescence (maximum emission wavelength: 660 nm), so it can be used as a probe to quantitatively detect Zn2+ in the range of 0-10 μM, with a detection limit as low as 4.8 nM, showing the probe has high sensitivity for Zn2+. And the probe has a fast response time to Zn2+ (less than 30 s) and a large Stoke-shift (179 nm). In addition, the high recovery rates in practical water samples, and the clear fluorescent images in living A549 cells were obtained, which are of great significance for the detection of Zn2+ in the environment and biosystem. Due to its simple operation, good selectivity and anti-interference ability, short detection time and high sensitivity, this probe has great application potential as a fast detection tool for Zn2+ in environmental water and biological samples.


[Functional role of several groups of neurons in oculomotor nerve nuclei (experimental study)].
The localization and functional significance of some groups of neurons in the main nuclei of the cat aculomotor nerve were studied with the aid of the method of retrograde degeneration. The topography of the oculomotor nerve nuclei in some representatives of the order of rodents, hare-like and predatory animals was specified. The conducted study demonstrated that the cell groups in the main nuclei of the oculomotor nerve were situated rostro-caudally in the following order: for the superior straight, internal straight and inferior straight muscles. In the middle and caudal sections of the nuclei there is a semidecussation.


The prognostic and predictive roles of plasma C-reactive protein and PD-L1 in non-small cell lung cancer.
Anti-PD-(L)1 agents have revolutionized the treatment paradigms of non-small cell lung cancer (NSCLC), while predictive biomarkers are limited. It has been previously shown that systemic inflammation, indicated by elevated C-reactive protein (CRP) level, is associated with a poor prognosis in anti-PD-(L)1 treated. The aim of the study was to analyze the prognostic and predictive value of CRP in addition to traditional prognostic and predictive markers and tumor PD-L1 score. We identified all NSCLC patients (n = 329) who had undergone PD-L1 tumor proportion score (TPS) analysis at Oulu University Hospital 2015-22. CRP levels, treatment history, immune checkpoint inhibitor (ICI) therapy details, and survival were collected. The patients were categorized based on CRP levels (≤10 vs. >10) and PD-L1 TPS scores (<50 vs. ≥50). In the whole cohort (n = 329), CRP level of ≤10 mg/L was associated with improved survival in univariate (HR 0.30, Cl 95% 0.22-0.41) and multivariate analyzes (HR 0.44, CI 95% 0.28-0.68). With ICI treated (n = 70), both CRP of ≤10 and PD-L1 TPS of ≥50 were associated with improved progression-free survival (PFS) in univariate (HR 0.51, CI 95% 0.27-0.96; HR 0.54, CI 95% 0.28-1.02) and multivariate (HR 0.48, CI 95% 0.26-0.90; HR 0.50, CI 95% 0.26-0.95) analyzes. The combination (PD-L1 TPS ≥50 and CRP >10) carried a high negative predictive value with a median PFS of 4.11 months (CI 95% 0.00-9.63), which was similar to patients with low PD-L1 (4.11 months, CI 95% 2.61-5.60). Adding plasma CRP levels to PD-L1 TPS significantly increased the predictive value of sole PD-L1. Furthermore, patients with high CRP beard little benefit from anti-PD-(L)1 therapies independent of PD-L1 score. The study highlights the combined evaluation of plasma CRP and PD-L1 TPS as a negative predictive marker for ICI therapies.


Cell-free DNA in the urine of rats exposed to ionizing radiation.
Investigation of cell-free DNA (cf-DNA) in body fluids, as a potential biomarker for assessing the effect of ionizing radiation on the organism, is of considerable interest. We investigated changes in the contents of cell-free mitochondrial DNA (cf-mtDNA) and cell-free nuclear DNA (cf-nDNA) in the urine of X-ray-exposed rats. Assays of cf-mtDNA and cf-nDNA were performed by a real-time PCR in rat urine collected before and after irradiation of animals with doses of 3 and 5 Gy. We also determined the presence of mutations in urine cf-mtDNA, as recognized by Surveyor nuclease. A sharp increase in cf-mtDNA and cf-nDNA in the urine of irradiated rats was observed within 24 h after exposure, followed by a decrease to normal levels. In all cases, the contents of cf-mtDNA fragment copies (estimated by gene tRNA) were significantly higher than those of cf-nDNA estimated by gene GAPDH. A certain portion of mutant cf-mtDNA fragments was detected in the urine of exposed rats, whereas they were absent in the urine of the same animals before irradiation. These preliminary data also suggest that the increased levels of urine cf-mtDNA and cf-nDNA may be a potential biomarker for noninvasive assessment of how the organism responds to ionizing radiation influence.


Interaction between renin and the autonomic nervous system in hypertension.
The abnormal distribution of plasma renin values described in established essential hypertension are also found in patients with very early, borderline hypertension. In established hypertension, renin values have been used to draw inferences about the pathophysiology of blood pressure elevation. Within this concept, the low-renin state is considered a volume-dependent (volume expanded) form of hypertension. The high-renin state is viewed as high-resistance hypertension caused by a renin-dependent vasoconstriction. However, the pathophysiology of high- and low-renin borderline hypertension does not follow the prediction from the volume-vasoconstriction theory. The high-renin state is often associated with an increase in cardiac output and normal values of vascular resistance. Even when the cardiac output is normal and the total peripheral resistance is elevated in high renin, the vasoconstriction is not renin-angiotensin dependent. The high-renin borderline and mild hypertension is a state of generalized, increased, sympathetic drive to the heart, blood vessels, and kidneys. After the influence of the autonomic nervous system is removed by pharmacologic blockade, blood pressure in patients with high-renin values becomes normal. To the contrary, pharmacologic antagonization of angiotensin II with a converting enzyme inhibitor does not lead to normal blood pressure values in patients with high-renin. Patients with borderline hypertension with low renin have normal plasma and blood volume values. However, because of decreased compliance of the peripheral capacitance space, the blood volume is shifted from the peripheral to the central (cardiopulmonary) portion of the circulation.(ABSTRACT TRUNCATED AT 250 WORDS)


Preterm Neurodevelopmental Outcomes Following Orosensory Entrainment Intervention.
Previous research indicates that the NTrainer, a pressurized pacifier programmed to produce pulsed pneumotactile stimulation during gavage feeds, has been found to facilitate non-nutritive suck development and shorten the length of hospital stay when used in the Neonatal Intensive Care Unit (NICU). Four groups of children, including infants of diabetic mothers (IDM), healthy controls (HI), and those with respiratory distress syndrome (RDS), or chronic lung disease (CLD), were randomly assigned to an NTrainer therapy or sham 'control' condition when in the NICU. At 30 months of age, 113/223 study participants were assessed using standardized language, motor, and cognitive assessments. No significant group differences were evident between the NTrainer and sham groups in language, motor, or cognitive functioning. The NTrainer did not improve nor adversely impact language, cognition, or motor outcomes.


IL-10 promoter single nucleotide polymorphisms are significantly associated with resistance to leprosy.
The minor haplotype -3575A/-2849G/-2763C in IL-10 promoter has been defined as a marker of disease resistance to leprosy and its severity in Brazilian population. Our investigation of six single-nucleotide polymorphisms (SNPs) in IL-10 promoter in 282 Indian leprosy patients and 266 healthy controls by direct PCR sequencing, however, showed that the extended haplotype: -3575T/-2849G/-2763C/-1082A/-819C/-592C was associated with resistance to leprosy per se and to the development of severe form of leprosy, using either a binomial (controls vs cases, P=0.01, OR=0.58, CI=0.37-0.89) or ordinal (controls vs paucibacillary vs multibacillary, P=0.004) model. Whereas, IL-10 haplotype -3575T/-2849G/-2763C/-1082A/-819T/-592A was associated with the risk of development of severe form of leprosy (P=0.0002) in contrast to the minor risk haplotype -3575T/-2849A/-2763C in the Brazilian population. The role of IL-10 promoter SNPs in Brazilian and Indian population strongly suggests the involvement of IL-10 locus in the outcome of leprosy.


Signal transfer from GPCRs to G proteins: role of the G alpha N-terminal region in rhodopsin-transducin coupling.
Catalysis of nucleotide exchange in heterotrimeric G proteins (Galphabetagamma) is a key step in cellular signal transduction mediated by G protein-coupled receptors. The Galpha N terminus with its helical stretch is thought to be crucial for G protein/activated receptor (R(*)) interaction. The N-terminal fatty acylation of Galpha is important for membrane targeting of G proteins. By applying biophysical techniques to the rhodopsin/transducin model system, we studied the effect of N-terminal truncations in Galpha. In Galphabetagamma, lack of the fatty acid and Galpha truncations up to 33 amino acids had little effect on R(*) binding and R(*)-catalyzed nucleotide exchange, implying that this region is not mandatory for R(*)/Galphabetagamma interaction. However, when the other hydrophobic modification of Galphabetagamma, the Ggamma C-terminal farnesyl moiety, is lacking, R(*) interaction requires the fatty acylated Galpha N terminus. This suggests that the two hydrophobic extensions can replace each other in the interaction of Galphabetagamma with R(*). We propose that in native Galphabetagamma, these two terminal regions are functionally redundant and form a microdomain that serves both to anchor the G protein to the membrane and to establish an initial docking complex with R(*). Accordingly, we find that the native fatty acylated Galpha is competent to interact with R(*) even in the absence of Gbetagamma, whereas nonacylated Galpha requires Gbetagamma for interaction. Experiments with N-terminally truncated Galpha subunits suggest that in the second step of the catalytic process, the receptor binds to the alphaN/beta1-loop region of Galpha to reduce nucleotide affinity and to make the Galpha C terminus available for subsequent interaction with R(*).


PVP-iodine in hydrosomes and hydrogel--a novel concept in wound therapy leads to enhanced epithelialization and reduced loss of skin grafts.
Moist wound treatment improves healing at a possibly increased risk of bacterial infection and many local antiseptics impair healing. A moist treatment modality with efficient antimicrobial activity would be desirable. In this monocentric, randomized, observer blinded, phase III study, a new hydrosome polyvinyl-pyrrolidone (PVP)-iodine preparation in hydrogel containing iodine in a 3% concentration (Repithel) was investigated for its effect on epithelialization in patients receiving meshed skin grafts. Grafts of 167 patients (donor site defects, burn wounds, or chronic defects) were dressed either with Repithel (n=83) covered with a gauze (Jelonet), or Jelonet-gauze only (n=84) until healing. Grafts receiving Repithel healed significantly earlier (9.4 days versus 12.4 days; p<0.0001) and faster than controls as measured by neo-epithelialization of mesh holes between days 7 and 11 (91.2+/-22.8% versus 82.3%+/-28.6, p<0.0001). A subgroup analysis showed that the effects on grafted burn wounds (p=0.0042) and chronic defects (p<0.0001) was more significant than on donor sites. Also a higher take rate of grafts (p=0.0053) and a reduced loss of grafts was observed with Repithel treatment (8 grafts versus 20 grafts) (p=0.0063, respectively). Smokers had improved graft take (p=0.0069) and higher rate of epithelialization (p=0.0040) compared to smokers of the control group. The results demonstrate significant clinical advantages of Repithel. This new local wound healing drug combines antisepsis and wound moisture efficiently resulting in significantly enhanced epithelialization, decreased transplant losses, and significantly improved healing especially in smokers.


Who benefited from the New Rural Cooperative Medical System in China? A case study on Anhui Province.
The goal of the New Rural Cooperative Medical System (NCMS) is to decrease the financial burden and improve the health of rural areas. The purpose of the present study is to determine how government subsidies vary between poorer and wealthier groups, especially in low-income regions in rural China. The distribution, amount, and equity of government subsidies delivered via NCMS to rural residents at different economic levels were assessed using benefit-incidence analysis, concentration index, Kakwani index, Gini index, Lorenz curve, and concentration curve. Household and health institution surveys were conducted in 2010, covering 9701 residents. Household socio-economic status, healthcare costs, out-of-pocket payments, and utilization information were collected in household interviews, and reimbursement policy was provided by institutional survey. The government subsidy concentration index was -0.055 for outpatients and 0.505 for inpatients; and the outpatient and inpatient subsidy Kakwani indexes were -0.376 and 0.184, respectively. The poorest 20 % of populations received 3.4 % of the total subsidy output; while the wealthiest 20 % received 54.3 %. The results showed that the distribution of outpatient subsidies was equitable, but the hospital subsidies disproportionally benefited wealthier people. Wealthier people benefited more than poorer people from the NCMS in terms of inpatient and total subsidies. For outpatients, the subsidies were unrelated to ability to pay. This contradicts the common belief that the NCMS does not exacerbate benefit inequity. Long-term policy is required to tackle this problem, specifically of redesign the NCMS reimbursement system.


Discovering dominant tumor immune archetypes in a pan-cancer census.
Cancers display significant heterogeneity with respect to tissue of origin, driver mutations, and other features of the surrounding tissue. It is likely that individual tumors engage common patterns of the immune system-here "archetypes"-creating prototypical non-destructive tumor immune microenvironments (TMEs) and modulating tumor-targeting. To discover the dominant immune system archetypes, the University of California, San Francisco (UCSF) Immunoprofiler Initiative (IPI) processed 364 individual tumors across 12 cancer types using standardized protocols. Computational clustering of flow cytometry and transcriptomic data obtained from cell sub-compartments uncovered dominant patterns of immune composition across cancers. These archetypes were profound insofar as they also differentiated tumors based upon unique immune and tumor gene-expression patterns. They also partitioned well-established classifications of tumor biology. The IPI resource provides a template for understanding cancer immunity as a collection of dominant patterns of immune organization and provides a rational path forward to learn how to modulate these to improve therapy.


A prevalence of posttransplantation cancers compared with cancers in people with human immunodeficiency virus/acquired immunodeficiency syndrome after highly active antiretroviral therapy.
Organ transplant recipients and human immunodeficiency virus & acquired Immunodeficiency Syndrome (HIV/AIDS) patients have immune deficiencies that are possible mechanisms to develop malignancy. The type of cancers associated with these 2 conditions might elucidate this premise. Our aim was to compare prevalence and type of cancers between kidney transplant recipients and patients with HIV/AIDS. We retrospectively reviewed 344 patients who underwent kidney transplantation from 1973 to 2007 compared them with 863 subjects with HIV/AIDS at the HIV-Netherlands/Australia/Thailand Research Collaboration (HIV-NAT) from 1997 to 2007. AIDS-defining cancers were excluded from the analysis. We compared the relative tumor risk with the age- and gender- matched general population of metropolitan Bangkok. The overall cancer risk for kidney transplant recipients (standardized incidence ratio [SIR] = 4.21) was comparable with HIV-infected patients (SIR = 3.88). Uroepithelial cancer was the most prevalent type in kidney transplant recipients, whereas cervical cancer was the most common malignancy in HIV-infected patients. The risks of developing hepatoma and non-Hodgkin's lymphoma were comparable between the groups. Kidney transplant recipients and HIV-infected patients show increased overall risks of certain types of cancers.


A comparative study of attitudes of the key relatives towards 'schizophrenic patients' and 'patients of disturbed family'.
Forty six key relatives of 'Schizophrenic Patients' who were hospitalised in Department of Psychiatry, K. G's, Medical College, Lucknow were administered 'Attitude Questionnaire' which covered five variables : Number of critical comments about someone else in the home ; Hostility; Dissatisfaction; Warmth and Emotional Involvement. Forty one key relatives of 'Disturbed Family's Patients' matched for sex and residence, were assessed on similar parameters. There was significant difference only in the area of 'Dissatisfaction'.


In vivo ¹H-NMR microimaging during seed imbibition, germination, and early growth.
Magnetic resonance imaging (MRI) is a superior noninvasive diagnostic tool widely used in clinical medicine, with more than 60 million MRI tests performed each year worldwide. More specialized high-resolution MRI systems capable of a resolution that is 100-1,000 times higher than standard MRI instruments are used primarily in materials science, but are used with increasing frequency in plant physiology. We have shown that high-resolution (1)H-nuclear magnetic resonance (NMR) microimaging can provide a wealth of information about the internal anatomy of plant seeds as small as 1 mm or even smaller. This chapter covers the methods associated with these imaging techniques in detail. We also discuss the application of (1)H-NMR microimaging to study in vivo seed imbibition, germination, and early seedling growth.


Reconsidering Donor Race in Predicting Allograft and Patient Survival Among Kidney Transplant Recipients.
Donor race should not be used in models to predict allograft and patient survival.Removing donor race from the Kidney Donor Risk Index may reduce kidney discard by reclassifying approximately 50% of high kidney donor profile index kidneys.Future prediction models should focus on using relevant biologic factors rather than social constructs when trying to predict outcomes.


Role of CFTR and anion exchanger in bicarbonate fluxes in C127 cell lines.
C127 cell lines transfected with wtCFTR, delta F508CFTR or vector were employed to determine HCO3- fluxes in the presence or absence of functional CFTR, using the pH-sensitive dye BCECF. Both cytosolic alkalinization and acidification were due to activity of anion exchanger and were similar in the three cell lines, indicating that expression of CFTR did not influence anion exchanger activity. In C127wt cells only, cAMP elevating agents significantly stimulated HCO3- fluxes, insensitive to the inhibitor of anion exchanger 4,4'-diisothiocyanate dihydrostilbene-2,2'-disulfonic acid, suggesting that activated CFTR directly mediates both HCO3- influx and efflux and therefore can contribute to intracellular and extracellular pH regulation.


MR imaging of the rotator cuff interval.
The rotator cuff interval is defined as the space between the anterior aspect of the supraspinatus tendon and the superior aspect of the subscapularis tendon. Knowledge of the anatomy, an understanding of the commonly encountered pathology, and an approach for the systematic inspection of the rotator cuff interval is crucial for the accurate characterization and diagnosis of pathology of this region. This article reviews the basic normal anatomy of the rotator cuff interval, imaging considerations unique to this area, and commonly encountered pathology.


Microvascular decompression for elderly patients with trigeminal neuralgia.
Microvascular decompression (MVD) has been demonstrated to be an excellent surgical treatment approach in younger patients with trigeminal neuralgia (TN). However, it is not clear whether there are additional morbidity and mortality risks for MVD in the elderly population. We performed a systematic literature review using six electronic databases for studies that compared outcomes for MVD for TN in elderly (cut-off ⩾60, 65, 70years) versus younger populations. Outcomes examined included success rate, deaths, strokes, thromboembolism, meningitis, cranial nerve deficits and cerebrospinal fluid leaks. There were 1524 patients in the elderly cohort and 3488 patients in the younger cohort. There was no significant difference in success rates in elderly versus younger patients (87.5% versus 84.8%; P=0.47). However, recurrence rates were lower in the elderly (11.9% versus 15.6%; P=0.03). The number of deaths in the elderly cohort was higher (0.9% versus 0.1%; P=0.003). Rates of stroke (2.5% versus 1%) and thromboembolism (1.1% versus 0%) were also higher for elderly TN patients. No differences were found for rates of meningitis, cranial nerve deficits or cerebrospinal fluid leak. MVD remains an effective and reasonable strategy in the elderly population. There is evidence to suggest that rates of complications such as death, stroke, and thromboembolism may be significantly higher in the elderly population. The presented results may be useful in the decision-making process for MVD in elderly patients with TN.


Stratum corneum levels of inflammatory mediators and natural moisturizing factor in patch test reactions to thiurams and fragrances and their possible role in discrimination between irritant and allergic reactions to hapten mixtures.
Patch test (PT) reactions to thiuram mix (TM) and fragrance mix (FM) I or II without concomitant reactions to their single constituents are potentially caused by the irritant properties of the mixes. Comparing inflammatory profiles of PT reactions to TM, FM I, FM II, and their constituents and assessing their potential in discrimination of irritant and allergic reactions. Levels of 14 cytokines and natural moisturizing factor (NMF) were determined in stratum corneum samples collected from PT reactions to TM, FM I or II, their constituents, and petrolatum (pet.) control sites in 36 individuals. Levels of interleukin (IL)-16, chemokine (CXC motif) ligand (CXCL) 8, CXCL10, chemokine (CC motif) ligand (CCL) 17, and CCL22 were significantly increased in reactions (+, ++) to thiurams and fragrances compared to their petrolatum. controls, except for PT reactions to FM I/II with negative breakdown testing in which, however, decreased levels of NMF were observed. In doubtful reactions to FM I/II with negative breakdown testing, NMF was significantly lower than in petrolatum controls. PT reactions to thiurams and fragrances indicate a Th2-skewed inflammation. The inflammatory profiles suggest that weak or doubtful FM I/II reactions without accompanying reaction to a constituent were irritant. IL-16 might be suitable to distinguish irritant from allergic reaction.


Preventing cancer.
Advancements in management protocols and chemotherapeutics have improved outcomes for patients diagnosed with cancer. Cancer, however, continues to claim many lives annually in the United States and around the world. There is a large body of evidence that is strong and consistent that through modification of diet and lifestyle habits, cancer can be a preventable disease. This article discusses these changes and highlights the evidence for and against implementing them.


Classic disease modifying anti-rheumatic drugs (DMARDs) in combination with infliximab. The Finnish experience.
To assess the performance of infliximab in a clinical setting, 364 rheumatoid arthritis (RA) patients from the National Register of Biological Treatment in Finland (ROB-FIN) were analysed. Corticosteroid usage and dose diminished (p<0.05 and 0.001, respectively) in patients on infliximab, of whom 51% also used one, 28% two and 16% three other concomitant DMARDs. A 34% of the RA patients used methotrexate+/-corticosteroids without any other DMARD. Methotrexate was most frequently used with sulphasalazine and/or hydroxychloroquine. Non-methotrexate patients most frequently used leflunomide or azathioprine combined with corticosteroids. The clinical effect of these combinations was similar to that of infliximab with methotrexate alone. The results indicate that infliximab can be used together with other DMARDs than methotrexate alone, quite according to the philosophy of the combination drug therapy, as the effectiveness is as good as or even slightly better than that of methotrexate and infliximab.


Concerning the thermal stability of E. coli 23S ribosomal RNA.
Total RNA prepared from E. coli by several extraction procedures behaves as a mixture of covalently continuous heat stable 23S, 16S and 4-5S components. 16S rRNA remains heat stable after isolation from such preparations, whereas isolated 23S rRNA is heat labile but becomes heat stable after EDTA treatment. This and other evidence leads to the conclusion that heat lability of purified 23S rRNA is due, not to nuclease contamination of the type observed in earlier studies of the stability of this RNA, but to polyvalent cation catalyzed temperature-dependent scission of phosphodiester bonds. Heat stability of 23S rRNA in total RNA is due to the presence in these preparations of a contaminant which appears to act as a chelator of polyvalent cations. This material is similar or identical to the pyrogenic E. coli lipopolysaccharide described by Westphal and coll.


Control of macrophage metabolism and activation by mTOR and Akt signaling.
Macrophages are pleiotropic cells that assume a variety of functions depending on their tissue of residence and tissue state. They maintain homeostasis as well as coordinate responses to stresses such as infection and metabolic challenge. The ability of macrophages to acquire diverse, context-dependent activities requires their activation (or polarization) to distinct functional states. While macrophage activation is well understood at the level of signal transduction and transcriptional regulation, the metabolic underpinnings are poorly understood. Importantly, emerging studies indicate that metabolic shifts play a pivotal role in control of macrophage activation and acquisition of context-dependent effector activities. The signals that drive macrophage activation impinge on metabolic pathways, allowing for coordinate control of macrophage activation and metabolism. Here we discuss how mTOR and Akt, major metabolic regulators and targets of such activation signals, control macrophage metabolism and activation. Dysregulated macrophage activities contribute to many diseases, including infectious, inflammatory, and metabolic diseases and cancer, thus a better understanding of metabolic control of macrophage activation could pave the way to the development of new therapeutic strategies.


[The use of computerized tomography in the detection of intra-orbital foreign bodies].
Use of the CT scanner as an instrument for foreign body detection and localization was explored under experimental conditions. The results show that a sufficiently dense radiopaque foreign body as small as 0.7mm3 can be detected and localized by presently available scanners.


Diagnostic categorization according to the First International Workshop on Ocular Sarcoidosis (FIWOS) criteria in a series of 11 patients.
Categorization of patients diagnosed with ocular sarcoidosis during the period 2009-2014. The medical records of patients with ocular sarcoidosis were reviewed and variables were collected to categorize the patients according to the criteria of the FIWOS. We found 11 patients, 7 women and 4 men, with sarcoid uveitis; the median age was 58 years. Bilateral panuveitis was the most common pattern (54.5%), followed by chronic anterior uveitis (27.2%). The diagnosis of sarcoidosis was definitive in 4 patients (36.3%), presumed in 5 (45.4%), probable in 1 (9%) and possible in 1 (9%). Ocular sarcoidosis was diagnosed in more than half of the patients who had no confirmatory biopsy. Bilateral panuveitis and chronic anterior uveitis were the patterns most frequently observed.


[Not Available].
98 wheet samples harvested in 1998 from three different experimental plots in saxony where investigated on the species of fusarium and the mycotoxins DON, ZEA and nivalenol (NIV). Analysis of DON was done by fast-Elisa-test and HPLC-MS. There was a good correlation between the two methods but a tendency of higher values determined by Elisa. NIV was also determined by HPLC-MS and ZEA by HPLC-fluorescence.


Nek1 Regulates Rad54 to Orchestrate Homologous Recombination and Replication Fork Stability.
Never-in-mitosis A-related kinase 1 (Nek1) has established roles in apoptosis and cell cycle regulation. We show that human Nek1 regulates homologous recombination (HR) by phosphorylating Rad54 at Ser572 in late G2 phase. Nek1 deficiency as well as expression of unphosphorylatable Rad54 (Rad54-S572A) cause unresolved Rad51 foci and confer a defect in HR. Phospho-mimic Rad54 (Rad54-S572E), in contrast, promotes HR and rescues the HR defect associated with Nek1 loss. Although expression of phospho-mimic Rad54 is beneficial for HR, it causes Rad51 removal from chromatin and degradation of stalled replication forks in S phase. Thus, G2-specific phosphorylation of Rad54 by Nek1 promotes Rad51 chromatin removal during HR in G2 phase, and its absence in S phase is required for replication fork stability. In summary, Nek1 regulates Rad51 removal to orchestrate HR and replication fork stability.


Comment on "Genomic epidemiology of superspreading events in Austria reveals mutational dynamics and transmission properties of SARS-CoV-2".
A reanalysis of SARS-CoV-2 deep sequencing data from donor-recipient pairs indicates that transmission bottlenecks are very narrow (one to three virions).


Evaluation of an in silico cardiac safety assay: using ion channel screening data to predict QT interval changes in the rabbit ventricular wedge.
Drugs that prolong the QT interval on the electrocardiogram present a major safety concern for pharmaceutical companies and regulatory agencies. Despite a range of assays performed to assess compound effects on the QT interval, QT prolongation remains a major cause of attrition during compound development. In silico assays could alleviate such problems. In this study we evaluated an in silico method of predicting the results of a rabbit left-ventricular wedge assay. Concentration-effect data were acquired from either: the high-throughput IonWorks/FLIPR; the medium-throughput PatchXpress ion channel assays; or QSAR, a statistical IC50 value prediction model, for hERG, fast sodium, L-type calcium and KCNQ1/minK channels. Drug block of channels was incorporated into a mathematical differential equation model of rabbit ventricular myocyte electrophysiology through modification of the maximal conductance of each channel by a factor dependent on the IC50 value, Hill coefficient and concentration of each compound tested. Simulations were performed and agreement with experimental results, based upon input data from the different assays, was evaluated. The assay was found to be 78% accurate, 72% sensitive and 81% specific when predicting QT prolongation (>10%) using PatchXpress assay data (77 compounds). Similar levels of predictivity were demonstrated using IonWorks/FLIPR data (121 compounds) with 78% accuracy, 73% sensitivity and 80% specificity. QT shortening (<-10%) was predicted with 77% accuracy, 33% sensitivity and 90% specificity using PatchXpress data and 71% accuracy, 42% sensitivity and 81% specificity using IonWorks/FLIPR data. Strong quantitative agreement between simulation and experimental results was also evident. The in silico action potential assay demonstrates good predictive ability, and is suitable for very high-throughput use in early drug development. Adoption of such an assay into cardiovascular safety assessment, integrating ion channel data from routine screens to infer results of animal-based tests, could provide a cost- and time-effective cardiac safety screen.


Prognosis and treatment of acute hydrocephalus following aneurysmal subarachnoid haemorrhage.
The aim of this study was to determine the rate of acute hydrocephalus in patients with aneurysmal subarachnoid haemorrhage (SAH) and evaluate its clinical prognosis and treatment options. We recruited 152 patients who had ruptured cerebral aneurysms between 2006 and 2010. Sequential CT scans were performed 24-72 hours after haemorrhage to examine the development of acute hydrocephalus following aneurysmal SAH. If hydrocephalus was present but the patient's condition was stable, the patient was closely monitored for 48 hours before undergoing a repeat CT scan to evaluate ventricular changes. To exclude aneurysm rebleeding, repeat CT scans were performed frequently in patients who continued to deteriorate clinically, and external ventricular drainage was performed if the deterioration was due to worsening hydrocephalus. In clinically stable patients with persistent hydrocephalus and patients with external ventricular drainage, lamina terminalis fenestration was performed during aneurysm clipping, and the procedure's therapeutic efficacy was evaluated at the postoperative follow-up. The overall incidence of acute hydrocephalus following aneurysmal SAH was 17.8% (27/152). Of these patients, 29.6% (eight of 27) recovered spontaneously within 48 hours of hydrocephalus development, 37.0% (10/27) required external ventricular drainage, and 33.3% (nine of 27) remained clinically stable with persistent hydrocephalus during the observational period. Hydrocephalus was significantly alleviated in eight of 11 patients who underwent lamina terminalis fenestration, and no related complications were observed during follow-up. Three patients, for whom intraoperative fenestration was ineffective, underwent shunt surgery. Our results showed approximately 30% of patients with acute hydrocephalus following aneurysmal SAH recover spontaneously. In the majority of patients with persistent hydrocephalus, lamina terminalis fenestration performed during aneurysm clipping is effective. Ventriculoperitoneal shunting can be considered in the remaining patients.


Acute organophosphorus toxicity in a regional hospital in Johannesburg, South Africa: A retrospective chart review.
Intentional and accidental organophosphorus exposures pose a significant healthcare-related burden on South African communities. This study will review the demographics, characteristics and clinical course of patients presenting with features of acute organophosphorus toxicity to a regional Emergency Centre in Johannesburg, South Africa. This was a retrospective chart review of all patients treated for possible acute organophosphorus toxicity from January 2020 to August 2021. A total of 205 patients were identified of which 134 patients were included in the study. The median age was 26 years with a male predominance (male= 56%, female=44%). 109 patients (81.3%) survived, 18 patients (13.4%) demised and the outcome of 7 patients (5.2%) was unknown. The median hospital length of stay was 8 days, (IQR= 5-13 days), and the longest hospital stay was 37 days in ICU. Atropinisation dose was significantly higher for intubated patients (median=140.0mg; IQR=90mg-219.5mg) compared to patients who were not intubated (median=60mg; IQR=20.5mg-120mg, p < 0.05). The length of stay was significantly higher for intubated patients (median=11 days; IQR=7-15 days) compared to patients who were not intubated (median=5 days; IQR=3-8 days, p < 0.00). There was a moderate positive correlation between atropinisation dose and length of stay (Correlation coefficient = 0.37, p < 0.00). There was a moderate negative correlation between atropinisation dose and cholinesterase level (Correlation coefficient= - 0.39, p < 0.00). Of those reported to have adverse effects 78.6%, were related to atropine toxicity. Our study shows a high mortality rate secondary to organophosphorus toxicity. Significant exposures and thus higher doses of atropine were associated with increased length of stay and need for intubation. We found a high incidence of atropine-related adverse effects. More studies are needed to further establish the balance between the therapeutic and adverse effects of high-dose atropine as a treatment modality for organophosphorus toxicity.


XAS Analysis of Reactions of (Arylimido)vanadium(V) Dichloride Complexes Containing Anionic NHC That Contains a Weakly Coordinating B(C6F5)3 Moiety (WCA-NHC) or Phenoxide Ligands with Al Alkyls: A Potential Ethylene Polymerization Catalyst with WCA-NHC Ligands.
(Arylimido)vanadium(V) dichloride complexes containing anionic N-heterocyclic carbene (NHC) ligands that contain weakly coordinating B(C6F5)3 moieties (WCA-NHC) of the type [V(NAr)Cl2(WCA-NHC-Ar')] (5, Ar = 2,6-Me2C6H3, Ar' = 2,6- i Pr2C6H3) showed significant catalytic activity for ethylene polymerization in the presence of Al cocatalysts (MAO and Al i Bu3); the activity by the 5-MAO catalyst (19 500 kg-PE/mol-V·h; TOF 11 600 min-1) is the highest among those reported using the other (imido)vanadium(V) complexes in the presence of MAO, and the 5-Al i Bu3 catalyst showed higher activity (66 000 kg-PE/mol-V·h; TOF 39 200 min-1). The V K-edge X-ray absorption near-edge structure (XANES) analyses (in toluene) strongly suggest the formation of certain vanadium(III) species by reduction with Al i Bu3 accompanying structural changes; the EXAFS analysis suggests the presence of the arylimido ligand and one V-Cl bond (2.34 ± 0.04 Å), which is longer than those [2.1901(8)-2.2462(8) Å] in the reported (imido)vanadium(V) complexes. The XANES analysis of [V(NAr)Cl2(OAr)] strongly suggests the formation of the other vanadium(III) species by reduction with Me2AlCl or Et2AlCl, and the EXAFS analysis suggests the presence of the arylimido ligand and two V-Cl bonds (2.45 ± 0.03 Å). The XANES spectra showed no significant changes in both the pre-edge peak(s) and the edge peak when these complexes were treated with MAO, suggesting that the basic geometry and the high oxidation state were preserved under these conditions.


[Transformation of an afferent tactile signal into a motor command in the cat motor cortex].
The activity of 112 units of the cat precruciate motor cortex was studied simultaneously with the performance of a placing reaction of the foreleg to tactile stimulation of its distal parts. The response latency to the tactile stimulation of the foreleg was 30-40 ms for the electromyographic response of m. biceps brachii and 20 +/- 10 ms for the earliest-responses of the motor cortical neurons. The electrical reaction of m. biceps brachii occured 5-10 ms after the termination of the electrical stimulation of the cortex by a puls series (the series duration was 25 ms, the pulse duration 0.2 ms, frequency 400 per s). Two different types of excitatory reactions of the motor cortex units were found: the sensory (s) ones occured at each tactile stimulus and did not depend on the presence or absence of the foreleg motion; the motor (m) type of reactions developed simultaneously with the motor placing reaction of the foreleg only. The minimal latency of cortical m- and s-type discharge was 20 +/- 10 ms. Electrical stimulation (5-25 micronA) through the microelectrode placed in the locus of neuronal activity which increased during active flexion of the foreleg, provoked in 70% of cases the electrical response in m. biceps brachii.


The United Kingdom National External Quality Assessment Scheme (blood transfusion laboratory practice): trends in proficiency and practice between 1985 and 2000.
Proficiency in blood transfusion laboratory practice has improved over the last 15 years (1985-2000). Error rates for ABO grouping have fallen from 0 x 19 to 0 x 02% (P = 0 x 003). A similar trend is evident for antibody screening, with error rates for false negative antibody screens falling from 3 x 2 to 0 x 5% (P < 0 x 001), and for antibody identification, for sera containing a single alloantibody, with error rates falling from 8 x 8% to 0 x 9% over the last 10 years (P < 0 x 001). Proficiency in serological crossmatching to detect clinically significant non-ABO incompatibilities (other than Kidd), has also improved (P < 0 x 001). However, error rates for RhD grouping have not changed and there has been a recent decline in proficiency in detecting weak ABO incompatibilities and Kidd antibodies with heterozygous cells. Procedures for pretransfusion testing have also been rationalized over this time. The indirect antiglobulin test (IAT) is used in isolation by 73% of laboratories for antibody screening, and 10% rely on the direct room temperature (DRT), immediate spin (IS) crossmatch in the absence (current and/or historical) of clinically significant antibodies. Despite the improvements in error rates that have occurred alongside the rationalizations, there is still evidence of noncompliance with current published BCSH guidelines and manufacturers' instructions.


Neural variability in premotor cortex is modulated by trial history and predicts behavioral performance.
In the study of decision making, emphasis is placed on different forms of perceptual integration, while the influence of other factors, such as memory, is ignored. In addition, it is believed that the information underlying decision making is carried in the rate of the neuronal response, while its variability is considered unspecific. Here we studied the influence of recent experience on motor decision making by analyzing the activity of neurons in the dorsal premotor area of two monkeys performing a countermanding arm task. We observe that the across-trial variability of the neural response strongly correlates with trial history-dependent changes in reaction time. Using a theoretical model of decision making, we show that a trial history-monitoring signal can explain the observed behavioral and neural modulation. Our study reveals that, in the neural processes that culminate in motor plan maturation, the evidence provided by perception and memory is reflected in mean rate and variance respectively.


Structural states and transitions of carp hemoglobin.
The wide ligand affinity range previously observed for carp hemoglobin is bounded at both extremes by regions of constant affinity. Within these regions, pH, organic phosphates, and the extent of ligand binding have no effect on the measured affinity and the cooperativity of ligand binding is greatly reduced or absent. The rates of CO recombination to fully and partially unliganded carp hemoglobin, under various organic phosphate and pH conditions, are shown to reflect this behavior. Constant kinetic rates are seen to directly correspond to the regions of constant affinity. Therefore, these are taken to be single protein conformations, one of high and one of low ligand affinity. In the simplest view, these conformations represent the R and T states of a two-state model, and most of the properties of carp hemoglobin are explained quite well within this framework. Increases in either hydrogen or phosphate ion concentrations favor the stabilization of the low affinity structure of even fully liganded carp hemoglobin. We have studied the structural transition from high to low affinity by monitoring the absorption spectra of carp hemoglobins at constant pH as a function of organic phosphate concentration. We find that different spectra are induced in both carp methemoglobin and cyanomethemoglobin by inositol hexaphosphate addition. Furthermore, the dependence of the magnitude of the spectral changes on pH and organic phosphate concentration is the close agreement with that predicted from studies of the ligand binding properties of the molecule.


Isolation and anticancer activity evaluation of rare Bisaryl anthraquinone antibiotics from novel Streptomyces sp. strain of NW Himalayan region.
Biosynthesis of bisaryl preanthraquinone antibiotics by various microorganisms differs in monomeric subunits as well as their dimerization positions leading to different configurations. The present study relates to the production of rare bisaryl anthraquinone antibiotics by a new Streptomyces strain isolated from Shivalik region of NW Himalayas. In vitro anticancer and anti-migratory effects of Setomimycin (9,9' bisanthraquinone antibiotic) was seen with a significant reduction in the expression of both MEK as well as ERK pathways in a dose dependent manner at 6.5 μM & 8 μM concentration in HCT-116 and 5.5 μM & 7 μM concentration in MCF-7 cells. In vivo studies in aggressive orthotopic mouse mammary carcinoma model (4T1) demonstrated about 76% reduction of primary tumor weight and 90.5% reduction in the tumor volume within two weeks. In vivo pharmacokinetics study of setomimycin revealed that it can be rapidly absorbed with an adequate plasma exposure and half-life which can be linked to its in vivo efficacy.


[Effect of movement arousal and hypoxia on circardian blood pressure in hypertensive patients with obstructive sleep apnea syndrome].
To investigate whether repeated arousals and hypoxia at the end of apnea contribute to nocturnal and diurnal elevation of blood pressure in obstructive sleep apnea syndrome (OSAS) patients with hypertension. 30 OSAS patients with hypertension and 30 OSAS patients without hypertension were studied. Movement arousal index (MAI), cortical arousal index (CAI), length of time during which nocturnal oxygen saturation decreased below 90% (OLT90%), apnea/hypopnea index (AHI), 24 h average systolic and diastolic blood pressure (ASBP and ADBP) were measured. Multiple stepwise regression analysis was performed to find the correlation between 24 h ASBP, ADBP and MAI, CAI, OLT90%, AHI, age, body mass index in OSAS patients with hypertension. MAI made the most significant contribution to 24 h ASBP and ADBP in OSAS patients with hypertension (ASBP: r = 0.485, P < 0.01; ADBP: r = 0.407, P < 0.05). OLT90% was the second most important contributing factor (ASBP: r = 0.382, P < 0.05; ADBP: r = 0.369, P < 0.05). Movement arousal and hypoxia may make an important contribution to circardian elevation of blood pressure in OSAS patients with hypertension. Both of them may contribute to the development of nocturnal and diurnal hypertension in patients with OSAS.


Flow dynamics of human sickle erythrocytes in the mesenteric microcirculation of the exchange-transfused rat.
To analyze the microvascular rheology of sickle cells in an intact animal model, rats were isovolemically exchange transfused with human normal (hemoglobin AA) or sickle (hemoglobin SS) erythrocytes (blood group O) or autologous red cells under ambient conditions, and the effects of the heterologous or autologous cells on (a) hemodynamics and respiration, (b) blood gases, and (c) acid-base status of the recipients were determined. Exchange transfusion of rats with autologous red cells or hemoglobin AA or hemoglobin SS erythrocytes was associated with stable mean arterial blood pressure, central venous pressure, respiration rate, blood pH, pCO2, and pO2 during the experimental period, except for tachycardia among the group of rats that received HbSS cells. Arteriovenous oxygen content varied among the three groups of animals, but, nonetheless, suggested adequate tissue oxygen supply under the conditions of the study. Acid-base status also was similar in the three groups of rats. The exchange-transfused rats were utilized to investigate the flow dynamics of red cells in the mesenteric microcirculation by applying intravital microscopy. Time-averaged velocities of the autologous red cells in 16- to 30-microns (id) vessels ranged from 1.07 to 1.25 mm/sec in single unbranched arterioles with varying flux and wall shear rates. Time-averaged velocities of the HbAA cells in single 15- to 35-microns arterioles ranged from 1.16 to 1.24 mm/sec with wall shear rates similar to the estimates for the autologous cells. For both rat and human HbAA RBCs, the flow dynamics were indicative of normal shear-dependent and deformability characteristics of the cells under the flow conditions. Sickle cells exhibited time-averaged velocities of 0.384 to 0.452 mm/sec, lower wall shear rates in 10- to 35-microns single unbranched arterioles, and three times less volumetric flux. In some arterioles, sickle cells with high axial ratio and low deformability showed definite adhesion to the endothelial surface, residing at such sites for several seconds until dislodged by the force of flow. Within single unbranched vessels or at microvascular bifurcations, sickle elliptocytes and sickle echinocytes with low deformability and high axial ratio obstructed flow and exhibited residence times of 2 to 88 sec, thereby causing stasis. These data illustrate the microvascular flow behavior of sickle cells and demonstrate the rheological disequilibrium state that can result as sickle cells course through successive segments of the microcirculation.


Sorafenib in the treatment of patients with hepatocellular carcinoma (HCC) and microvascular infiltration: a systematic review and meta-analysis.
Microvascular invasion is shown to be an independent risk factor for liver cancer recurrence. Timely treatment may reduce the recurrence rate and prolong total survival time. The aim of this study was to investigate the effectiveness of sorafenib in treating patients with hepatocellular carcinoma (HCC) and microvascular invasion. A comprehensive literature search was conducted in PubMed, EMBASE, MEDLINE, web of science and Cochrane Library databases for articles published up to December 2019. Two researchers independently reviewed and cross-checked independent reports with sufficient information. A meta-analysis was conducted to assess the impact of sorafenib on mortality in patients with HCC and microvascular involvement. Four studies were included in the qualitative and quantitative analyses, comprising 955 cancer events and 505 cancer deaths. Meta-analyses showed that sorafenib treatment was associated with an improved survival rate versus no sorafenib treatment in patients with HCC and microvascular invasion (relative risk 1.369, 95% confidence interval 1.193, 1.570). Sorafenib treatment may improve survival in patients with HCC and microvascular invasion. However, due to the potential for residual confounding, the results should be interpreted with caution.


Circulating CD8+CD28- suppressor T cells tied to poorer prognosis among metastatic breast cancer patients receiving adoptive T-cell therapy: A cohort study.
This study aimed to determine the prognostic value of circulating CD8+CD28- T lymphocytes among breast cancer patients treated with adoptive T-lymphocyte immunotherapy after chemotherapy. Two hundred and thirty-two breast cancer patients underwent adoptive T-cell immunotherapy. Circulating CD8+CD28- proportion was measured by flow cytometry. Median proportion of CD8+CD28- was 24.2% and set as the categorical cutoff value for further analysis. The median survival was estimated by Kaplan-Meier curve, with difference detection and hazard ratio estimation by log-rank test and Cox hazard proportion regression model. With adoptive T-cell therapy, patients with higher CD8+CD28- levels experienced median progression-free and overall survival of 7.1 months and 26.9 months, respectively-significantly shorter than patients with lower levels (11.8 and 36.2 months). CD8+CD28- proportion >24.2% demonstrated a hazard ratio (HR) of 2.06 (95% confidence interval [CI] 1.31-3.12) for progression and an HR of 1.97 (95% CI 1.06-3.67) for death. Among patients who had received previous first-line chemotherapy, CD8+CD28- proportion >24.2% demonstrated an HR of 2.66 (95% CI 1.45-4.88) for progression. Among patients exposed to previous second-line or higher chemotherapy, CD8+CD28- proportion >24.2% demonstrated a 486% higher risk for death (HR = 5.86, 95% CI 1.77-19.39). A 1% increase in suppressive T cells was associated with a 5% increased risk of death. Elevated peripheral blood CD8+CD28- was associated with poorer prognosis for metastatic breast cancer, especially for higher risk of progression among patients with first-line chemotherapy and higher risk of death among patients with more than second-line chemotherapy.


Stress influenced increase in phenolic content and radical scavenging capacity of Rhodotorula glutinis CCY 20-2-26.
Rhodotorula glutinis CCY 20-2-26 when grown under controlled stress of either NaCl (1-5 %) or H2O2 (1-5 mM) on basal media exhibited a twofold increase in its total phenolic contents. The radical scavenging capacities (RSCs) as determined by ABTS test were found to be highest in 4 mM H2O2 (1.44 mM TEAC mg-1) and 4 % NaCl (1.13 mM TEAC mg-1) as compared to control samples (0.41 mM TEAC mg-1). Similarly, the RSCs as determined by DPPH test were also highest in 4 % NaCl (1.83 mM TEAC mg-1) and 4 mM H2O2 (1.78 mM TEAC mg-1) compared to control (0.48 TEAC mg-1). The relative RSCs from EPR spin-trapping assay for H2O2-stressed cultures were highest in 1 mM H2O2 (56.1 μM TEAC g-1) whereas in NaCl-stressed cultures it was highest in 5 % NaCl (44.6 μM TEAC g-1) as compared to control (30.9 μM TEAC g-1). Five phenolic compounds (gallic acid, benzoic acid, catechin, caffeic acid and ferulic acid) were detected for the first time in R. glutinis CCY 20-2-26.


A study of the effect of mifepristone (antiprogesterone) followed by prostaglandin on uterine activity and fetal heart rate in patients having a termination of pregnancy.
In the 72 h after a single oral dose of 400 mg of the antiprogesterone mifepristone, 12 out of 14 first and one second trimester fetuses had a slight increase in heart rate; 2 fetuses died and one aborted. During the same 72 h, uterine activity increased moderately, and was physiological with no increase in resting pressure. The treatment sensitized the uterus to prostaglandin (PG) about ten-fold. A low, 0.05 mg IM, dose of sulprostone caused the demise of 5 more fetuses and caused the onset of clinical abortion in less than 2 h. After a relatively short hypertonic phase uterine resting pressure fell to normal levels and active contractions occurred leading to expulsion of uterine contents. The plasma level of progesterone (P) remained unaltered after mifepristone treatment, but the levels of estradiol 17b (E2) and cortisol increased. The plasma level of mifepristone was 1640 +/- 424 ng. ml -1 at 72 h, and the substance was still detectable after one week.


Determining Complete Chromosomal Haplotypes by mLinker.
Haplotype ("haploid genotype") phase is the combination of genotypes at sites of genetic variation along a chromosome [1]. We previously demonstrated that the complete chromosomal haplotype of diploid human genomes can be determined using molecular linkage from Hi-C sequencing and linked-reads sequencing [2]. In this chapter, we present a step-by-step guide to perform this analysis using mLinker, a software package for haplotype inference.


Enhanced Peripheral Chemoreflex Drive Is Associated with Cardiorespiratory Disorders in Mice with Coronary Heart Disease.
Coronary heart disease (CHD) is a prevalent cardiovascular disease characterized by coronary artery blood flow reductions caused by lipid deposition and oxidation within the coronary arteries. Dyslipidemia is associated with local tissue damage by oxidative stress/inflammation and carotid bodies (CB) peripheral chemoreceptors are heavily modulated by both reactive oxygen species and pro-inflammatory molecules (i.e., cytokines). Despite this, it is not know whether CB-mediated chemoreflex drive may be affected in CHD. In the present study, we evaluated peripheral CB-mediated chemoreflex drive, cardiac autonomic function, and the incidence of breathing disorders in a murine model of CHD. Compared to age-matched control mice, CHD mice showed enhanced CB-chemoreflex drive (twofold increase in the hypoxic ventilatory response), cardiac sympathoexcitation, and irregular breathing disorders. Remarkably, all these were closely linked to the enhanced CB-mediated chemoreflex drive. Our results showed that mice with CHD displayed an enhanced CB chemoreflex, sympathoexcitation, and disordered breathing and suggest that CBs may be involved in chronic cardiorespiratory alterations in the setting of CHD.


Establishment of a new method, transcription-reverse transcription concerted reaction, for detection of plasma hnRNP B1 mRNA, a biomarker of lung cancer.
Development of an early detection marker is one of the most important strategies for improving overall prognosis in lung cancer patients. We previously reported that hnRNP B1--an RNA binding protein--is overexpressed in lung cancer tissue from the early stage of cancer, and found that hnRNP B1 mRNA is detectable in the plasma of lung cancer patients using real-time RT-PCR. The purpose of this study was to establish a quick and simple method for detecting plasma hnRNP B1mRNA for use in screening for lung cancer. TRC, a homogenous method for fluorescence real-time monitoring of isothermal RNA amplification using intercalation activating fluorescence DNA probe, was used to detect plasma hnRNP B1 mRNA. The detection limit of hnRNP B1 mRNA by TRC using synthetic control RNA or total RNA derived from a lung cancer cell line was 25 or 8.65 x 10(2) copies, respectively. Using total RNA extracted from 600 mul of plasma, we detected hnRNP B1 mRNA in 39.1% (9/23) of lung cancer patients, with levels ranging from 1.9 to 19,045.5 copies/100 ng RNA, and in 5.2% (5/97) of healthy volunteers. Copy numbers were not associated with age, gender, smoking status, or histological type of cancer. TRC could detect 10(3) copies of hnRNP B1 mRNA in 10 min. Detection of plasma hnRNP B1 mRNA by TRC is a quick, easy, and non-invasive method suitable for lung cancer screening.


Evans' syndrome as a presenting manifestation of atypical paroxysmal cold hemoglobinuria.
Paroxysmal cold hemoglobinuria is a rare and potentially life-threatening acquired hemolytic anemia occurring either as an acute transient anemia following several different viral syndromes, or in a chronic idiopathic form. Episodic hemolysis in paroxysmal cold hemoglobinuria is usually associated with a biphasic (Donath-Landsteiner) IgG cold-reactive complement-fixing autohemolysin with anti-P specificity. Paroxysmal cold hemoglobinuria has not previously been associated with malignancy nor has it been clearly shown to be steroid-responsive. This report describes a patient with steroid-responsive autoimmune hemolytic anemia and immune thrombocytopenia (Evans' syndrome) associated with oat cell carcinoma of the lung and a unique biphasic anti-IgM autohemolysin. This case extends the spectrum of biphasic antibody-mediated immune cytopenias and widens both the clinical and the serologic definition of paroxysmal cold hemoglobinuria.


Assessment of microRNA profiles in small extracellular vesicles isolated from bovine colostrum with different immunoglobulin G concentrations.
The consumption of bovine colostrum by newborn calves during the first days of life is essential to ensure the transfer of passive immunity. In addition to critical IgG, colostrum also contains non-IgG biomolecules, including microRNA (miRNA). The present study investigated the profiles of miRNA in small extracellular vesicles (sEV) isolated from bovine colostrum with high (256.5 ± 5.7 mg/mL, mean ± standard deviation, n = 4) and low (62.8 ± 3.6 mg/mL, n = 4) concentrations of IgG. Different combination of sEV extraction methods and bioinformatic pipelines (miRDeep2 and sRNAbench) for miRNA analysis were evaluated. Results showed that miRCURY exosome Cell/Urine/CSF and miRNeasy Mini kits yielded the highest RNA concentration. The miRNA-seq data analysis showed miRDeep2 yielded more comprehensive miRNAome compared with sRNAbench (527 versus 392 unique miRNA), whereas 389 shared miRNA were identified using both approaches. The profiles of top 50 miRNA were the same using both approaches, and their abundance contributed to 91.7% and 94.3% of total abundance of miRNA using miRDeep2 and sRNAbennch, respectively. These core miRNA were predicted to target 2,655 genes, which regulate 78 KEGG (Kyoto Encyclopedia of Genes and Genomes) level-3 pathways including PI3K-Akt and MAPK signaling pathway, axon guidance, and focal adhesion. The expression profiles of sEV-associated miRNA were similar between high- and low-IgG colostrum samples, despite the fact that the abundance of miR-27a-3p was higher in colostrum with high concentrations of IgG. In conclusion, a core miRNAome in bovine colostrum may play a role in regulating health and developmental stages in neonatal calves, independent of IgG concentration.


Motion-Dependent Effects of Functional Magnetic Resonance Imaging Preprocessing Methodology on Global Functional Connectivity.
Background: While functional magnetic resonance imaging (fMRI) has become an established noninvasive tool for studying brain activity in both healthy and diseased states, no broad consensus has been reached regarding preprocessing methodology. Furthermore, the relationship between variations in preprocessing and functional connectivity (FC) networks remains incompletely understood. Purpose: The aim of this study was to relate FC to (1) choices in preprocessing methodology and (2) subject motion. Methods: Clinical and MRI data were analyzed from healthy subjects acquired as part of the Autism Brain Imaging Data Exchange (ABIDE). Data were obtained from 508 healthy subjects. Data from subjects in the highest and lowest quartiles for motion were used to calculate the interaction between motion and preprocessing. Data were analyzed across four domains of fMRI preprocessing: (1) pipeline, (2) global signal regression (GSR), (3) bandpass filtering, and (4) anatomic atlas. For the FC network calculated from each preprocessing scheme, overall FC using Pearson correlation, as well as leaf fraction and diameter, was calculated for each subject, and statistical comparison was made across schemes using generalized estimating equations. Results: FC and global network properties were significantly affected by each preprocessing step, and each preprocessing step significantly interacted with subject motion to differentially affect global functional network properties, with GSR having the strongest effect. Conclusion: Preprocessing choices in fMRI studies influence overall FC and global network properties and can have motion-dependent effects. Impact statement Different parameter choices across the four domains of functional magnetic resonance imaging (fMRI) preprocessing analyzed in this study (global signal regression, bandpass filtering, anatomic atlas, and pipeline) were associated with differences in functional connectivity (FC) as well as global network properties in healthy subjects. Some fMRI preprocessing parameter choices resulted in motion-dependent effects on FC and global network properties.


Misdiagnosis of Myasthenia Gravis and Subsequent Clinical Implication: A case report and review of literature.
The autoimmune disease, myasthenia gravis (MG), can mimic a variety of neurological disorders leading to a delay in diagnosis and treatment. On occasions, misdiagnosis of MG could lead to unnecessary and potentially harmful therapeutic interventions. We report on a 12 year-old boy, in whom MG was mistaken for meningitic sequelae and subsequently for critical neuropathy/myopathy resulting in considerable morbidity for nearly a decade. Subsequent correct diagnosis and optimal management resulted in significant improvement in his functional status. We discuss the importance of considering MG as one of the potential differential diagnoses among cases of recurrent respiratory pump failure, or unexplained bulbar symptoms where documentary proof of the previous diagnoses including work-up for MG is lacking. We also review the literature on MG misdiagnosis and highlight the potential pitfalls in MG diagnosis.


A new alternative to cosmetics preservation and the effect of the particle size of the emulsion droplets on preservation efficacy.
This study describes a new alternative combination preservative containing caprylyl glycol, phenethyl alcohol and glyceryl caprylate and investigates the effects of the particle size of the emulsion droplet on the anti-microbial activity of the said blend in the formulation. The anti-microbial activity of the ingredients and the blend were determined via MIC, MBC, MFC and fractional inhibitory concentration (FIC) determination using the broth microdilution method. The blend inhibited the micro-organism contamination of the oil-in-water emulsion, and its performance was affected by the particle size of the emulsion droplet, as shown by the in vitro microbial challenge test. Results show that the MIC/MBC/MFC values of the blend were lower than those of any of the ingredients used alone. Any two of the ingredients presented no antagonistic activities against all the tested micro-organisms, and synergism or additive effects were also observed. The challenge test also showed that the action of the blend against bacteria and yeast was only 0.3% and 0.2%, respectively, and mould was completely inhibited at 1.4%, meeting the requirements of the PCPC and European Pharmacopoeia. The results showed that anti-microbial activity was gradually enhanced when the particle size of the emulsion droplets increased significantly in the range of 100-900 nm. A positive correlation was found between anti-microbial activity and particle size. The synergistic performance of caprylyl glycol, phenethyl alcohol and glyceryl caprylate and the anti-microbial activity of their blend suggest that their combination is effective and exhibits broad-spectrum anti-microbial activity. Furthermore, the results show a positive correlation between the anti-microbial activity of the preservative and the particle size of the emulsion droplets in the range of 100-900 nm, when the same concentration of the blend is used in the same formulation. The particle size of the emulsion droplets is demonstrated to be a newly discovered factor that influences the preservation of cosmetic products.


In vivo production of interleukin-10 by non-T cells in rheumatoid arthritis, Sjögren's syndrome, and systemic lupus erythematosus. A potential mechanism of B lymphocyte hyperactivity and autoimmunity.
Interleukin-10 (IL-10) is a potent stimulator of B lymphocytes in vitro. In vivo dysregulation of IL-10 gene expression was therefore analyzed in patients with rheumatoid arthritis (RA), primary Sjögren's syndrome (SS), and systemic lupus erythematosus (SLE). Spontaneous production of IL-10 by peripheral blood mononuclear cells was measured using reverse transcription polymerase chain reaction and enzyme-linked immunosorbent assay in untreated patients with either RA (n = 10), SS (n = 10), or SLE (n = 10), and in 15 normal control subjects. IL-10 production was dramatically higher in RA, SS, and SLE patients than in controls. In each group, both B lymphocytes and monocytes, but not T lymphocytes, produced IL-10. IL-10 production is increased in RA, SS, and SLE. It may play a role in B lymphocyte hyperactivity and in the development of autoimmunity.


Catecholamine stimulation is associated with impaired myocardial O(2) utilization in heart failure.
To investigate the effect of alpha,beta(1) and beta(2) adrenergic receptor (AR) stimulation on coronary hemodynamics, myocardial oxygen consumption (M(v)O(2)) and metabolic substrate preference in advanced dilated cardiomyopathy (DCM). We studied 19 conscious, instrumented dogs with pacing-induced DCM. We evaluated systemic, coronary hemodynamics and M(v)O(2) in response to norepinephrine (NOR, 0.05-0.4 microg/kg per min), dobutamine (DOB, 1-10 microg/kg per min), phenylephrine (PHE, 1-5 microg/kg per min) and isoproterenol (ISO, 0.05-0.4 microg/kg per min) alone or in the presence of metoprolol (ISO+MET). Experiments were conducted in control state and in advanced DCM, 4-5 weeks after the initiation of pacing. Contractile responses (LV dP/dt) to catecholamines were desensitized and accompanied by a parallel decrease in heart rate-adjusted myocardial O(2) consumption (M(v)O(2/beat)), when alpha(PHE) or beta(1) (DOB) or both alpha/beta(1) (NOR) AR were stimulated in DCM. This was due to impaired transmyocardial (Ao-Cs) O(2) extraction rather than limitations in CBF responses. There was an associated shift in myocardial metabolism, evidenced by an increased preference for glycolytic substrates (Respiratory Quotient) following administration of any of these three adrenergic agonists in DCM. Combined beta(1)/beta(2) stimulation with ISO or beta(2)-AR stimulation (ISO+MET) in DCM resulted in greater M(v)O(2/beat), [(Ao-Cs) O(2)] extraction, and decreases in myocardial RQ consistent with a shift toward oxidation of FFA. The impairment in contractile responses to dobutamine and norepinephrine in DCM is associated with impaired myocardial O(2) extraction, and a shift toward a preference for glycolysis. A different myocardial metabolic pattern suggestive of increased oxidation of FFA with increased myocardial O(2) extraction was observed in the presence of combined beta(1)/beta(2) stimulation with isoproterenol or beta(2) stimulation (ISO+MET). These data suggest that beta(2)-AR stimulation in DCM shifts substrate preference toward FFA oxidation associated with greater M(v)O(2) requirements. These findings identify a putative metabolic effect of beta(2) -AR in DCM that may be deleterious.


Isolation and sequencing of rat liver bilirubin UDP-glucuronosyltransferase cDNA: possible alternate splicing of a common primary transcript.
A 1763-bp cDNA for rat liver bilirubin UDP-glucuronosyltransferase (UDPGT) was isolated. Bilirubin UDPGT activity was demonstrated by transfection of the pcDL1 vector carrying the cDNA into COS7 monkey kidney cells. The cDNA shares an identical 913-bp sequence (corresponding to the C-terminal 247 amino acid residues) with that for rat liver 3-methylcholanthrene-inducible 4-nitrophenol UDPGT including the locus where a -1 frameshift mutation was found in the 4-nitrophenol UDPGT cDNA from the jaundiced homozygous Gunn rat. The result suggests that both the UDPGTs are derived from a common primary-transcript and that the multiple defects of UDPGT isoenzymes observed in the homozygous Gunn rat may be produced by a single-mutated-locus after an alternative splicing of the 5' end region.


Optical waveguide lightmode spectroscopy technique-based immunosensor development for aflatoxin B1 determination in spice paprika samples.
Optical waveguide lightmode spectroscopy (OWLS) technique has been applied to label-free detection of aflatoxin B1 in a competitive immunoassay format, with the aim to compare the analytical goodness of the developed OWLS immunosenor with HPLC and enzyme-linked immunosorbent assay (ELISA) methods for the detection of aflatoxin in spice paprika matrix. We have also assessed applicability of the QuEChERS method prior to ELISA measurements, and the results were compared to those obtained by traditional solvent extraction followed by immunoaffinity clean-up. The AFB1 content of sixty commercial spice paprika samples from different countries were measured with the developed and optimized OWLS immunosensor. Comparing the results from the indirect immunosensor to that obtained by HPLC or ELISA provided excellent correlation (with regression coefficients above 0.94) indicating that the competitive OWLS immunosensor has a potential for quick determination of aflatoxin B1 in paprika samples.


Seasonal variation in the superficial vein thrombosis frequency.
A seasonal variation of venous thromboembolic disease frequency is subject to discussion, and has been recently suggested for superficial vein thrombosis (SVT) in a small retrospective study. Our aim was to search for a seasonal variation of SVT frequency according to the data of larger studies. We analyzed the data of three French prospective multicenter studies with different designs which have included patients with SVT (STENOX, POST, and STEPH studies). Seasonal variation of SVT frequency was evaluated by comparing the observed seasonal frequency of SVT to a theoretical frequency of 25% for each season. The analysis included 1395 patients and 4.75 seasonal cycles. The difference to a theoretical frequency of 25% was statistically significant in one study (POST, p = 0.044). The higher risk difference was -6.1% (95% CI -11.7–−0.5) in summer in STENOX, +7.1% (95% CI +2.7-+11.5) in winter in POST and 4.2% (95% CI -5.2-+13.7) in spring in STEPH, corresponding to a relative risk of 0.80, 1.40 and 1.20, respectively. A seasonal variation was found in only one study which has the weakest methodology to warrant completeness. Variation pattern was


Regio-controllable [2+2] benzannulation with two adjacent C(sp3)-H bonds.
Regiocontrol in traditional cycloaddition reactions between unsaturated carbon compounds is often challenging. The increasing focus in modern medicinal chemistry on benzocyclobutene (BCB) scaffolds indicates the need for alternative, more selective routes to diverse rigid carbocycles rich in C(sp3) character. Here, we report a palladium-catalyzed double C-H activation of two adjacent methylene units in carboxylic acids, enabled by bidentate amide-pyridone ligands, to achieve a regio-controllable synthesis of BCBs through a formal [2+2] cycloaddition involving σ bonds only (two C-H bonds and two aryl-halogen bonds). A wide range of cyclic and acyclic aliphatic acids, as well as dihaloheteroarenes, are compatible, generating diversely functionalized BCBs and hetero-BCBs present in drug molecules and bioactive natural products.


Cholesterol-related knowledge, attitudes, and behaviors in a low-income, urban patient population.
To determine knowledge, attitudes, behaviors, and self-reported cholesterol measurement in a low-income, urban patient population, we conducted an interview survey of users and potential users of primary care services in a public health care system for low-income Harris County, Texas, residents. The response rate was 93%, with a final sample of 547 randomly selected subjects 18 years of age and older, who were Hispanic (54%), black (28%), non-Hispanic white (14%), and Asian, Native American, or other (4%). Results indicated that 76% had heard of serum or blood cholesterol, and 30% reported past cholesterol measurement. Knowledge that dietary saturated fat can raise blood cholesterol ranged from 11% in Hispanic men to 51% in non-Hispanic white men and women. A lower percentage of Hispanics correctly answered all knowledge questions, and Hispanics reported higher-fat food choices than blacks and non-Hispanic whites. More than 90% of the respondents expressed interest in more information on diet, 60% reported that they read nutrition labels, and 15% said they have been trying to reduce blood cholesterol levels. A lower percentage of Hispanics reported previous cholesterol measurement than blacks or non-Hispanic whites, a difference that persisted after adjusting for multiple factors associated with cholesterol measurement. Older age (older than 50) and more physician visits in the past year also were associated with past cholesterol measurement. Comparisons with national surveys show that cholesterol knowledge and actual measurement in this low-income sample lag behind those of the national population. Yet, despite gaps in knowledge and cholesterol measurement, respondents showed positive attitudes about and interest in cholesterol-lowering interventions.