Expression of a gene family in the dimorphic fungus Mucor racemosus which exhibits striking similarity to human ras genes.
Sporulation, spore germination, and yeast-hypha dimorphism in the filamentous fungus Mucor racemosus provide useful model systems to study cell development in eucaryotic cells. Three RAS genes (MRAS1, MRAS2, and MRAS3) from M. racemosus have been cloned, and their nucleotide sequences have been determined. The predicted amino acid sequences and the sizes of the three MRAS proteins exhibit a high degree of similarity with other ras proteins, including that encoded by H-ras, which have been implicated in regulation of proliferation and development in eucaryotic cells by mediating signal transduction pathways. The MRAS proteins show conservation of functional domains proposed for ras proteins, including guanine nucleotide interaction domains, an effector domain, a binding epitope for neutralizing antibody Y13-259, and the COOH-terminal CAAX box, which is a site of thiocylation and membrane attachment. Amino acid sequences unique to each MRAS protein occur adjacent to the CAAX box, consistent with the location of the hypervariable region in other ras proteins. Northern (RNA) analysis was used to study expression of the three MRAS genes in relation to cell development. Gene-specific probes for two of these genes, MRAS1 and MRAS3, hybridized to different 1.3-kb mRNA transcripts. The accumulation of these transcripts depended on the developmental stage, and this pattern was different between the two MRAS genes. No transcript for MRAS2 was detected in the developmental stages examined. The unique patterns of MRAS transcript accumulation suggest that individual MRAS genes and proteins may play distinct roles in cell growth or development.


Multiple hormonal dysregulation as determinant of low physical performance and mobility in older persons.
Mobility-disability is a common condition in older individuals. Many factors, including the age-related hormonal dysregulation, may concur to the development of disability in the elderly. In fact, during the aging process it is observed an imbalance between anabolic hormones that decrease (testosterone, dehydroepiandrosterone sulphate (DHEAS), estradiol, insulin like growth factor-1 (IGF-1) and Vitamin D) and catabolic hormones (cortisol, thyroid hormones) that increase. We start this review focusing on the mechanisms by which anabolic and catabolic hormones may affect physical performance and mobility. To address the role of the hormonal dysregulation to mobility-disability, we start to discuss the contribution of the single hormonal derangement. The studies used in this review were selected according to the period of time of publication, ranging from 2002 to 2013, and the age of the participants (≥65 years). We devoted particular attention to the effects of anabolic hormones (DHEAS, testosterone, estradiol, Vitamin D and IGF-1) on both skeletal muscle mass and strength, as well as other objective indicators of physical performance. We also analyzed the reasons beyond the inconclusive data coming from RCTs using sex hormones, thyroid hormones, and vitamin D (dosage, duration of treatment, baseline hormonal values and reached hormonal levels). We finally hypothesized that the parallel decline of anabolic hormones has a higher impact than a single hormonal derangement on adverse mobility outcomes in older population. Given the multifactorial origin of low mobility, we underlined the need of future synergistic optional treatments (micronutrients and exercise) to improve the effectiveness of hormonal treatment and to safely ameliorate the anabolic hormonal status and mobility in older individuals.


The association of prenatal attachment and perinatal factors with pre- and postpartum depression in first-time mothers.
This prospective study investigated associations between prenatal attachment of adult first-time mothers to the unborn child, perinatal factors and levels of depression before and up to 18 months after delivery. Primiparas (N = 161) without specific risk factors answered the following questionnaires during the last term of pregnancy (t1): Edinburgh Postnatal Depression Scale (EPDS), Maternal Antenatal Attachment Scale (MAAS), questionnaire on the schema of the unborn child, and a questionnaire about the pregnancy. Perinatal data were taken from the patients' files. The EPDS was answered 3 weeks (t2, N = 157), 6 months (t3, N = 159), and 18 months (t4, N = 132) postpartum. During pregnancy, 16.9 % of the women indicated mild depressive symptoms, and 7.5 %, medium to severe symptoms of depression. Mild symptoms of depression were found in 25.5 % at t2, 10.1 % at t3, and 12.2 % at t4; medium to severe symptoms were reported by 7.6, 1.9 and 5.6 %, respectively. Women with low control during delivery (emergency Caesarean) showed a tendency for higher levels (p = 0.067) of depression at t3 than women with elective Caesarean did. The quality of prenatal attachment to the unborn child correlated negatively with depressive symptoms at t1-t4. The closer the prenatal attachment of a mother to her unborn child, the less symptoms of depression she reports during the last term of pregnancy and postpartum. Therefore, promoting good mother-child attachment during pregnancy might influence the level of postpartum depression.


Cardiopulmonary Resuscitation Capacity in Referral Hospitals in Nigeria: Understanding the Global Health Disparity in Resuscitation Medicine.
Little is known about the state of resuscitation services in low- and middle-income countries (LMICs), including Nigeria, Africa's most populous country. We sought to assess the cardiopulmonary resuscitation (CPR) care in referral hospitals across Nigeria to better inform capacity-building initiatives. We designed a survey to evaluate infrastructure, equipment, personnel, training, and clinical management, as no standardized instrument for assessing resuscitation in LMICs was available. We included referral teaching hospitals with a functioning intensive care unit (ICU) and a department of anaesthesiology. We pilot-tested our tool at four hospitals in Nigeria and recruited participants electronically via the Nigerian Society of Anaesthetists directory. Our survey included 17 hospitals (82% public, 12% private, 6% public-private partnership), although some questions include only a subset of these. We found that 20% (3 out of 15) of hospitals had a cardiac arrest response team system, 21% (3/14) documented CPR events, and 21% (3/14) reviewed such events for education and quality improvement. Most basic supplies were sufficient in the ICU (100% [15/15] availability of defibrillators, 94% [16/17] of adrenaline) but were less available in other departments. While 67% [10/15] of hospitals had a resuscitation training program, only 27% [4/15] had at least half their physicians trained in basic life support. In this first large-scale assessment of resuscitation care in Nigeria, we found progress in training centre development and supply availability, but a paucity of cardiac arrest response team systems. Our data indicate a need for improved capacity development, especially in documentation and continuous quality improvement, both of which are low-cost solutions.


Glutamatergic neurons in the medial prefrontal cortex mediate the formation and retrieval of cocaine-associated memories in mice.
In drug addiction, environmental stimuli previously associated with cocaine use readily elicit cocaine-associated memories, which persist long after abstinence and trigger cocaine craving and consumption. Although previous studies suggest that the medial prefrontal cortex (mPFC) is involved in the expression of cocaine-addictive behaviors, it remains unclear whether excitatory and inhibitory neurons in the mPFC are causally related to the formation and retrieval of cocaine-associated memories. To address this issue, we used the designer receptors exclusively activated by designer drugs (DREADD) technology combined with a cocaine-induced conditioned place preference (CPP) paradigm. We suppressed mPFC neuronal activity in a cell-type- and timing-dependent manner. C57BL/6J wild-type mice received bilateral intra-mPFC infusion of an adeno-associated virus (AAV) expressing inhibitory DREADD (hM4Di) under the control of CaMKII promotor to selectively suppress mPFC pyramidal neurons. GAD67-Cre mice received bilateral intra-mPFC infusion of a Cre-dependent AAV expressing hM4Di to specifically silence GABAergic neurons. Chemogenetic suppression of mPFC pyramidal neurons significantly attenuated both the acquisition and expression of cocaine CPP, while suppression of mPFC GABAergic neurons affected neither the acquisition nor expression of cocaine CPP. Moreover, chemogenetic inhibition of mPFC glutamatergic neurons did not affect the acquisition and expression of lithium chloride-induced conditioned place aversion. These results suggest that the activation of glutamatergic, but not GABAergic, neurons in the mPFC mediates both the formation and retrieval of cocaine-associated memories.


Palmitoyl protein thioesterase-1 deficiency impairs synaptic vesicle recycling at nerve terminals, contributing to neuropathology in humans and mice.
Neuronal ceroid lipofuscinoses represent the most common childhood neurodegenerative storage disorders. Infantile neuronal ceroid lipofuscinosis (INCL) is caused by palmitoyl protein thioesterase-1 (PPT1) deficiency. Although INCL patients show signs of abnormal neurotransmission, manifested by myoclonus and seizures, the molecular mechanisms by which PPT1 deficiency causes this abnormality remain obscure. Neurotransmission relies on repeated cycles of exo- and endocytosis of the synaptic vesicles (SVs), in which several palmitoylated proteins play critical roles. These proteins facilitate membrane fusion, which is required for neurotransmitter exocytosis, recycling of the fused SV membrane components, and regeneration of fresh vesicles. However, palmitoylated proteins require depalmitoylation for recycling. Using postmortem brain tissues from an INCL patient and tissue from the PPT1-knockout (PPT1-KO) mice that mimic INCL, we report here that PPT1 deficiency caused persistent membrane anchorage of the palmitoylated SV proteins, which hindered the recycling of the vesicle components that normally fuse with the presynaptic plasma membrane during SV exocytosis. Thus, the regeneration of fresh SVs, essential for maintaining the SV pool size at the synapses, was impaired, leading to a progressive loss of readily releasable SVs and abnormal neurotransmission. This abnormality may contribute to INCL neuropathology.


Molecular and cellular correlates of the CIITA-mediated inhibition of HTLV-2 Tax-2 transactivator function resulting in loss of viral replication.
MHC class II transactivator CIITA inhibits the function of HTLV-2 Tax-2 viral transactivator and, consequently, the replication of the virus in infected cells. Moreover overexpression of the nuclear factor NF-YB, that cooperates with CIITA for the expression of MHC class II genes, results also in inhibition of Tax-2 transactivation. The purpose of this investigation was to assess the cellular and molecular basis of the CIITA-mediated inhibition on Tax-2, and the relative role of NF-YB in this phenomenon. By co-immunoprecipitation of lysates from 293T cells cotransfected with CIITA or fragments of it, and Tax-2 it was assessed whether the two factors interact in vivo. A similar approach was used to assess Tax-2-NF-YB interaction. In parallel, deletion fragments of CIITA were tested for the inhibition of Tax-2-dependent HTLV-2 LTR-luciferase transactivation. Subcellular localization of CIITA and Tax-2 was investigated by immunofluorescence and confocal microscopy. CIITA and Tax-2 interact in vivo through at least two independent regions, at the 1-252 N-term and at the 410-1130 C-term, respectively. Interestingly only the 1-252 N-term region mediates Tax-2 functional inhibition. CIITA and Tax-2 are localized both in the cytoplasm and in the nucleus, when separately expressed. Instead, when coexpressed, most of Tax-2 colocalize with CIITA in cytoplasm and around the nuclear membrane. The Tax-2 minor remaining nuclear portion also co-localizes with CIITA. Interestingly, when CIITA nucleus-cytoplasm shuttling is blocked by leptomycin B treatment, most of the Tax-2 molecules are also blocked and co-localize with CIITA in the nucleus, suggesting that CIITA-Tax-2 binding does not preclude Tax-2 entry into the nucleus.Finally, the nuclear factor NF-YB, also strongly binds to Tax-2. Notably, although endogenous NF-YB does not inhibit Tax-2-dependent HTLV-2 LTR transactivation, it still binds to Tax-2, and in presence of CIITA, this binding seems to increase. These results strongly suggest that CIITA inhibit Tax-2 by binding the viral transactivator both directly or through a tripartite interaction with NF-YB in. CIITA is therefore a viral restriction factor for HTLV-2 and this open the possibility to control HTLV-2 viral replication and spreading by the controlled induction of CIITA in infected cells.


A discrete cis element in the human immunodeficiency virus long terminal repeat mediates synergistic trans activation by cytomegalovirus immediate-early proteins.
The major immediate-early (IE) promoter of human cytomegalovirus directs the expression of several differentially spliced and polyadenylated mRNAs that encode isoformic proteins with apparent molecular masses of 55, 72, and 86 kDa. All of these proteins are potent transcriptional regulatory proteins. We are interested in the collateral interactions between human cytomegalovirus and human immunodeficiency virus (HIV) in the context of dual infection of a cell. The roles of the specific IE protein isoforms and their respective response elements involved in trans activation of the HIV long terminal repeat (LTR) are not known. Here we present evidence that major IE proteins IE86, IE72, and IE55 are capable of trans-activating the HIV LTR in a T-cell line, HUT-78. The IE55 isoform noncooperatively stimulates the HIV LTR in the presence of either isoform IE72 or IE86. Interactions between isoforms IE72 and IE86, however, result in strong synergistic activation of the LTR. Our results suggest that a specific 155-amino-acid protein domain that is unique for the IE86 protein participates in this synergic interaction. Point mutational analysis of the LTR identified a distinct cis-acting target site, located between nucleotide positions -174 and -163, that mediates exclusively synergistic trans activation by the IE72 and IE86 proteins. Finally, this study underscores the role of a cellular intermediate(s) for communicating the synergic interactions between two IE trans activators.


Cortical plasticity of motor-eloquent areas measured by navigated transcranial magnetic stimulation in patients with glioma.
OBJECTIVE The goal of this study was to obtain a better understanding of the mechanisms underlying cerebral plasticity. Coupled with noninvasive detection of its occurrence, such an understanding has huge potential to improve glioma therapy. The authors aimed to demonstrate the frequency of plastic reshaping, find clues to the patterns behind it, and prove that it can be recognized noninvasively using navigated transcranial magnetic stimulation (nTMS). METHODS The authors used nTMS to map cortical motor representation in 22 patients with gliomas affecting the precentral gyrus, preoperatively and 3-42 months postoperatively. Location changes of the primary motor area, defined as hotspots and map centers of gravity, were measured. RESULTS Spatial normalization and analysis of hotspots showed an average shift of 5.1 ± 0.9 mm (mean ± SEM) on the mediolateral axis, and 10.7 ± 1.6 mm on the anteroposterior axis. Map centers of gravity were found to have shifted by 4.6 ± 0.8 mm on the mediolateral, and 8.7 ± 1.5 mm on the anteroposterior axis. Motor-eloquent points tended to shift toward the tumor by 4.5 ± 3.6 mm if the lesion was anterior to the rolandic region and by 2.6 ± 3.3 mm if it was located posterior to the rolandic region. Overall, 9 of 16 (56%) patients with high-grade glioma and 3 of 6 (50%) patients with low-grade glioma showed a functional shift > 10 mm at the cortical level. CONCLUSIONS Despite the small size of this series, analysis of these data showed that cortical functional reorganization occurs quite frequently. Moreover, nTMS was shown to detect such plastic reorganization noninvasively.


A prospective pilot study of intravitreal aflibercept for the treatment of chronic central serous chorioretinopathy: the CONTAIN study.
To evaluate the role of intravitreal aflibercept injection as a treatment for eyes with chronic central serous chorioretinopathy (CSCR). This prospective pilot study enrolled 12 patients with chronic CSCR who received a 6-month treatment regimen of intravitreal aflibercept. Patients were followed with monthly Early Treatment of Diabetic Retinopathy Study (ETDRS) best-corrected visual acuity (BCVA) and spectral domain optical coherence tomography (SD-OCT) with enhanced depth imaging. All patients were men between 29 and 64 years (median 55). Subfoveal fluid was present on OCT for a median duration of 6 months (range 4-29 months) prior to treatment. Baseline BCVA ranged from 20/25 to 20/160 (median 20/50) with a mean of 62 (SD=13) ETDRS letters. No patients experienced serious ocular or systemic adverse events over the course of the study. Post-treatment BCVA ranged from 20/20 to 20/200 (median 20/40), with a mean of 64 (SD=16) ETDRS letters (p=0.56). At baseline, three patients (25%) had BCVA of ≥20/40 versus seven patients (58%) at the conclusion of the study. Two patients gained at least 15 ETDRS letters and no patients lost more than 15 ETDRS letters. Six of 12 patients (50%) had complete resolution of subfoveal fluid. Mean central macular thickness decreased from 400 µm (SD=104 µm) to 306 µm (SD=94 µm) (p=0.03), and mean subfoveal fluid decreased from 159 µm (SD=93 µm) to 49 µm (SD=68 µm) (p=0.02). Mean choroidal thickness decreased from 307 µm (SD=72 µm) to 263 µm (SD=63 µm) (p=0.0003). Intravitreal aflibercept was well tolerated over a 6-month treatment course for chronic CSCR. No change was observed in visual acuity metrics. Anatomic trends may suggest some morphological activity, but larger controlled trials are needed. NCT01710332.


The effect of abortion on having and achieving aspirational one-year plans.
Women commonly report seeking abortion in order to achieve personal life goals. Few studies have investigated whether an abortion enables women to achieve such goals. Data are from the Turnaway Study, a prospective cohort study of women recruited from 30 abortion facilities across the US. The sample included women in one of four groups: Women who presented for abortion just over the facility's gestational limit, were denied an abortion and went on to parent the child (Parenting Turnaways, n = 146) or did not parent (Non-Parenting Turnaways, n = 64), those who presented just under the facility's gestational limit and received an abortion (Near-Limits, n = 413) and those who presented in the first trimester and received an abortion (First Trimesters, n = 254). Participants were interviewed by telephone one week, six months and one year after they sought an abortion. We used mixed effects logistic regression to assess the relationship between receiving versus being denied abortion and having an aspirational one year goal and achieving it. The 757 participants in this analysis reported a total of 1,304 one-year plans. The most common one-year plans were related to education (21.3 %), employment (18.9 %), other (16.3 %), and change in residence (10.4 %). Most goals (80 %) were aspirational, defined as a positive plan for the next year. First Trimesters and Near-Limits were over 6 times as likely as Parenting Turnaways to report aspirational one-year plans [Adjusted Odds Ratio (AOR) = 6.37 and 6.56 respectively, p < 0.001 for both]. Among all plans in which achievement was measurable (n = 1,024, 87 %), Near-Limits (45.6 %, AOR = 1.91, p = 0.003) and Non-Parenting Turnaways (47.9 %, AOR = 2.09, p = 0.026) were more likely to have both an aspirational plan and to have achieved it than Parenting Turnaways (30.4 %). These findings suggest that ensuring women can have a wanted abortion enables them to maintain a positive future outlook and achieve their aspirational life plans.


Effects of cholinergic blockade, adrenergic blockade and sympathetic denervation on gastrointestinal myoelectric activity in guinea pig.
Gastrointestinal myoelectric activity was recorded from conscious guinea pigs using sets of bipolar electrodes that had been surgically implanted onto the serosal surface of the gastric antrum or small intestine. The role of nerves in the control of the migrating myoelectric complex (MMC) was investigated using cholinergic and adrenergic receptor antagonists, guanethidine to block sympathetic transmission, reserpine to deplete intestinal monoamines and 6-hydroxydopamine (6-OHDA) to produce chemical sympathectomy and surgical sympathetic denervation of the stomach and small intestine. Subcutaneous infusion of hyoscine (1.0 or 3.0 mg/kg and 1.0 or 3.0 mg/kg/hr) blocked the initiation of phase 3 of the MMC. In the presence of hyoscine, intermittent electrical spiking comparable to that of phase 2 of the MMC was recorded. Hexamethonium infusion (3.0 or 10.0 mg/kg and 3.0 or 10.0 mg/kg/hr) also blocked the initiation of phase 3 and reduced the frequency of the intermittent spiking activity. Acute blockade of adrenergic receptors by simultaneous s.c. infusion of phentolamine and propranolol (1.0 mg/kg and 1.0 mg/kg/hr of each drug) did not alter the MMC, whereas infusion of guanethidine (3.75 or 7.5 mg/kg and 3.75 or 7.5 mg/kg/hr) increased the frequency of the MMC by significantly shortening the duration of phase 2. Sympathetic denervation of the stomach and small intestine did not block the MMC but decreased the cycle frequency by significantly prolonging the duration of phase 2. Chemical sympathectomy using 6-OHDA treatment (3 X 250 mg/kg s.c.) also did not abolish the MMC but decreased the frequency by increasing the duration of phase 2. Reserpine treatment (2.5 mg/kg s.c.) decreased the frequency of the MMC to a greater extent than that produced by 6-OHDA treatment. Histochemical and immunohistochemical evaluation of tissues taken from denervated, 6-OHDA- and reserpine-treated animals confirmed that surgical denervation removed sensory and adrenergic nerves to the stomach and small intestine, whereas chemical sympathectomy affected only adrenergic neurons. Reserpine treatment depleted norepinephrine from extrinsic adrenergic neurons and also depleted 5-hydroxytryptamine from intrinsic intestinal neurons. In the guinea pig, antral myoelectric activity is briefly inhibited during duodenal phase 3. Gastric inhibition persisted during duodenal phase 3 in surgically denervated animals, in 6-OHDA-treated animals and in animals treated acutely with guanethidine. These results indicate that extrinsic sensory and sympathetic nerves are not required for the initiation and propagation of the MMC in the guinea pig small intestine.(ABSTRACT TRUNCATED AT 400 WORDS)


Rotation of a single acetylene molecule on Cu(001) by tunneling electrons in STM.
We study the elementary processes behind one of the pioneering works on scanning tunneling microscope controlled reactions of single molecules [Stipe et al., Phys. Rev. Lett. 81, 1263 (1998)]. Using the Keldysh-Green function approach for the vibrational generation rate in combination with density functional theory calculations to obtain realistic parameters we reproduce the experimental rotation rate of an acetylene molecule on a Cu(100) surface as a function of bias voltage and tunneling current. This combined approach allows us to identify the reaction coordinate mode of the acetylene rotation and its anharmonic coupling with the C-H stretch mode. We show that three different elementary processes, the excitation of C-H stretch, the overtone ladder climbing of the hindered rotational mode, and the combination band excitation together explain the rotation of the acetylene molecule on Cu(100).


Synthesis and structures of lithium alkoxytris(dimethylphenylsilyl)borates.
The reactions of trialkyl borates B(OR)3 (R = Me, i-Pr) with dimethylphenylsilyllithium gave lithium alkoxytris(dimethylphenylsilyl)borates 1a,b. X-ray crystallographic analysis showed that 1a,b were obtained as a contact ion pair and a solvent-separated ion pair, respectively. The 11B and 29Si chemical shifts as well as the Si-B and B-O bond lengths of 1a,b and related compounds largely change, depending on the numbers of silyl and alkoxy substituents.


Quality Indicators of End-of-Life Care in Patients With Cancer: What Rate Is Right?
To develop data-driven and achievable benchmark rates for end-of-life quality indicators using administrative data from four provinces in Canada. Indicators of end-of-life care were defined and measured using linked administrative data for 33 health regions across British Columbia, Alberta, Ontario, and Nova Scotia. These were emergency department use, intensive care unit admission, physician house calls and home care visits before death, and death in hospital. An empiric benchmark was defined using indicator rates from the top-ranked regions to include the top decile of patients overall. Funnel plots were used to graph each region's age- and sex-adjusted indicator rates along with the overall rate and 95% confidence limits. Rates varied approximately two- to four-fold across the regions, with physician house calls showing the greatest variation. Benchmark rates based on the top decile performers were emergency department use, 34%; intensive care unit admission, 2%; physician house calls, 34%; home care visits, 63%; and death in hospital, 38%. With the exception of intensive care unit admission, funnel plots demonstrated that overall indicator rates and their confidence limits were uniformly worse than benchmarks even after adjusting for age and sex. Few regions met the benchmark rates. There is significant variation in end-of-life quality indicators across regions in four provinces in Canada. Using this study's methods-deriving empiric benchmarks and funnel plots-regions can determine their relative performance with greater context that facilitates priority setting and resource deployment. Applying this study's methods can support quality improvement by decreasing variation and striving for a target.


Identification and characterization of unrecognized viruses in stool samples of non-polio acute flaccid paralysis children by simplified VIDISCA.
The use of sequence independent methods combined with next generation sequencing for identification purposes in clinical samples appears promising and exciting results have been achieved to understand unexplained infections. One sequence independent method, Virus Discovery based on cDNA Amplified Fragment Length Polymorphism (VIDISCA) is capable of identifying viruses that would have remained unidentified in standard diagnostics or cell cultures. VIDISCA is normally combined with next generation sequencing, however, we set up a simplified VIDISCA which can be used in case next generation sequencing is not possible. Stool samples of 10 patients with unexplained acute flaccid paralysis showing cytopathic effect in rhabdomyosarcoma cells and/or mouse cells were used to test the efficiency of this method. To further characterize the viruses, VIDISCA-positive samples were amplified and sequenced with gene specific primers. Simplified VIDISCA detected seven viruses (70%) and the proportion of eukaryotic viral sequences from each sample ranged from 8.3 to 45.8%. Human enterovirus EV-B97, EV-B100, echovirus-9 and echovirus-21, human parechovirus type-3, human astrovirus probably a type-3/5 recombinant, and tetnovirus-1 were identified. Phylogenetic analysis based on the VP1 region demonstrated that the human enteroviruses are more divergent isolates circulating in the community. Our data support that a simplified VIDISCA protocol can efficiently identify unrecognized viruses grown in cell culture with low cost, limited time without need of advanced technical expertise. Also complex data interpretation is avoided thus the method can be used as a powerful diagnostic tool in limited resources. Redesigning the routine diagnostics might lead to additional detection of previously undiagnosed viruses in clinical samples of patients.


Bioactivity and applications of sulphated polysaccharides from marine microalgae.
Marine microalgae have been used for a long time as food for humans, such as Arthrospira (formerly, Spirulina), and for animals in aquaculture. The biomass of these microalgae and the compounds they produce have been shown to possess several biological applications with numerous health benefits. The present review puts up-to-date the research on the biological activities and applications of polysaccharides, active biocompounds synthesized by marine unicellular algae, which are, most of the times, released into the surrounding medium (exo- or extracellular polysaccharides, EPS). It goes through the most studied activities of sulphated polysaccharides (sPS) or their derivatives, but also highlights lesser known applications as hypolipidaemic or hypoglycaemic, or as biolubricant agents and drag-reducers. Therefore, the great potentials of sPS from marine microalgae to be used as nutraceuticals, therapeutic agents, cosmetics, or in other areas, such as engineering, are approached in this review.


Cannabis sativa L. as a Natural Drug Meeting the Criteria of a Multitarget Approach to Treatment.
Cannabis sativa L. turned out to be a valuable source of chemical compounds of various structures, showing pharmacological activity. The most important groups of compounds include phytocannabinoids and terpenes. The pharmacological activity of Cannabis (in epilepsy, sclerosis multiplex (SM), vomiting and nausea, pain, appetite loss, inflammatory bowel diseases (IBDs), Parkinson's disease, Tourette's syndrome, schizophrenia, glaucoma, and coronavirus disease 2019 (COVID-19)), which has been proven so far, results from the affinity of these compounds predominantly for the receptors of the endocannabinoid system (the cannabinoid receptor type 1 (CB1), type two (CB2), and the G protein-coupled receptor 55 (GPR55)) but, also, for peroxisome proliferator-activated receptor (PPAR), glycine receptors, serotonin receptors (5-HT), transient receptor potential channels (TRP), and GPR, opioid receptors. The synergism of action of phytochemicals present in Cannabis sp. raw material is also expressed in their increased bioavailability and penetration through the blood-brain barrier. This review provides an overview of phytochemistry and pharmacology of compounds present in Cannabis extracts in the context of the current knowledge about their synergistic actions and the implications of clinical use in the treatment of selected diseases.


Proteogenomic analysis reveals unanticipated adaptations of colorectal tumor cells to deficiencies in DNA mismatch repair.
A growing body of genomic data on human cancers poses the critical question of how genomic variations translate to cancer phenotypes. We used standardized shotgun proteomics and targeted protein quantitation platforms to analyze a panel of 10 colon cancer cell lines differing by mutations in DNA mismatch repair (MMR) genes. In addition, we performed transcriptome sequencing (RNA-seq) to enable detection of protein sequence variants from the proteomic data. Biologic replicate cultures yielded highly consistent proteomic inventories with a cumulative total of 6,513 protein groups with a protein false discovery rate of 3.17% across all cell lines. Networks of coexpressed proteins with differential expression based on MMR status revealed impact on protein folding, turnover and transport, on cellular metabolism and on DNA and RNA synthesis and repair. Analysis of variant amino acid sequences suggested higher stability of proteins affected by naturally occurring germline polymorphisms than of proteins affected by somatic protein sequence changes. The data provide evidence for multisystem adaptation to MMR deficiency with a stress response that targets misfolded proteins for degradation through the ubiquitin-dependent proteasome pathway. Enrichment analysis suggested epithelial-to-mesenchymal transition in RKO cells, as evidenced by increased mobility and invasion properties compared with SW480. The observed proteomic profiles demonstrate previously unknown consequences of altered DNA repair and provide an expanded basis for mechanistic interpretation of MMR phenotypes.


Estimating the Shear Resistance of Flocculated Kaolin Aggregates: Effect of Flocculation Time, Flocculant Dose, and Water Quality.
The resistance of kaolin aggregates to shearing in water clarification and recovery operations is a critical input in designing thickener feed wells. A recently formulated but already available criterion is used to determine the shear strength of flocculated kaolin aggregates. The flocculant is a high molecular weight anionic polyelectrolyte. The resistance of the aggregates is evaluated as a function of flocculation time, flocculant dosage, and water quality. The determination is based on a standardized experimental method. First, the time evolution of the average size of kaolin flocs is measured when aggregates are exposed to incremental shear rates from a predetermined base value. Then, the results are fitted to a pseudo-first-order model that allows deriving a characteristic value of the shear rate of rupture associated with the upper limit of the strength of the aggregates. In seawater, at a given dose of flocculant, the strength of the aggregates increases with time up to a maximum; however, at longer times, the resistance decreases until it settles at a stable value corresponding to stable aggregates in size and structure. A higher flocculant dosage leads to stronger aggregates due to more bridges between particles and polymers, leading to a more intricate and resistant particle network. In industrial water with very low salt content, the resistance of the kaolin aggregates is higher than in seawater for the same dose of flocculant. The salt weakens the resistance of the aggregates and works against the efficiency of the flocculant. The study should be of practical interest to concentration plants that use seawater in their operations.


Nonlinear metrics assessing motor variability in a standardized pipetting task: Between- and within-subject variance components.
The aim of this study was to estimate the between days test-retest reliability of nonlinear metrics used to quantify motor variability in a repetitive pipetting task. On three separate days, 14 healthy subjects performed pipetting as a general model of repetitive tasks. The task consisted of transferring liquid 20 times with a cycle time of 2.8s from a pickup tube to eight target tubes placed on a table in front of the subjects. The motion of hand, arm and the pipette tip was tracked in 3D and the shoulder elevation and elbow flexion angle were obtained. Motor variability was assessed using nonlinear metrics based on information theory and recurrence quantification analysis. Between- and within- (between-days) subject variance components were computed using a one-way random effect model, and intra-class correlation coefficients (ICC) were calculated from the variance components as standardized measures of reliability. Most of the metrics displayed a considerable between-days variance component and therefore the ICC showed a slight to moderate reliability. The reported data on between- and within-subject variability can be used to design future studies using nonlinear motor variability metrics on kinematics data.


Cryoprecipitate use in the PROMMTT study.
There are few clinical data to guide the use of cryoprecipitate in severely injured trauma patients. Cryoprecipitate is a rich source of fibrinogen and has been associated with improved survival in animal as well as limited human studies. Our objectives were to identify patterns and predictors of cryoprecipitate use and determine whether transfusing cryoprecipitate was associated with improved survival. This secondary analysis of 1,238 of 1,245 PRospective Observational Multicenter Major Trauma Transfusion (PROMMTT) study patients who had timed transfusion data included 359 (29%) who received cryoprecipitate. For this analysis, one dose of cryoprecipitate was defined as 10 U. Unadjusted predictors of cryoprecipitate use were identified using logistic regression. Multivariable time-dependent Cox models were performed to examine the association of cryoprecipitate on time to in-hospital death. Cryoprecipitate use varied significantly by center, ranging from 7% to 82%. Among patients who received cryoprecipitate, the median number of units infused by 24 hours was 10 (interquartile range, 10-20). The median time from admission to first cryoprecipitate unit was 2.7 hours (interquartile range, 1.7-4.4 hours). Of those who died of a hemorrhagic death within 6 hours of admission, 72% received no cryoprecipitate. Other unadjusted predictors of cryoprecipitate use included Injury Severity Score (ISS), initial fibrinogen levels, base deficit, international normalized ratio, prothrombin time/partial thromboplastin time, hemoglobin, damage-control surgery, and surgical intervention of the chest and abdomen. Cryoprecipitate use was not associated with in-hospital mortality after adjusting for initial pH, initial hemoglobin, emergency department systolic blood pressure, emergency department Glasgow Coma Scale (GCS) score, blood product use, ISS, and center. Ten US Level 1 trauma centers vary greatly in their timing and use of cryoprecipitate in severely injured trauma patients. We could not identify any association of cryoprecipitate use with in-hospital mortality, although most patients did not receive this product. Randomized controlled studies are needed to determine if cryoprecipitate (or fibrinogen concentrates) have a beneficial effect.


Western Australian pregnancy cohort (Raine) Study: Generation 1.
The purpose of the Raine Study is to improve human health and well-being by studying the life-course of a cohort of Western Australians, based on a life-course conceptual framework that considers interactions between genetics, phenotypes, behaviours, the environment and developmental and social outcomes. Between May 1989 and November 1991, 2900 pregnant women were enrolled in the Raine Study in Perth, Western Australia. In total, 2730 women gave birth to 2868 children (Generation 2) between August 1989 and April 1992. The mothers and fathers of Generation 2 are referred to as Generation 1 of the Raine Study. In the most recent Generation 1 follow-up, 636 mothers and 462 fathers participated. Until the 26-year follow-up of Generation 1 the focus of research within the Raine Study was on outcomes in Generation 2, with information on the parents mainly being used to examine its influence on their children's outcomes. For example, recent findings showed that several characteristics of mothers, such as obesity, early mid-gestational weight gain and socioeconomic status were associated with non-alcoholic fatty liver disease, adiposity and cardiometabolic characteristics in offspring. Other findings showed that parents with back pain were more likely to have offspring who experienced back pain. Also, non-linear and dynamic relationships were found between maternal working hours and offspring overweight or obesity. The Raine Study will continue to provide access to its dense longitudinal genetic, phenotypic, behavioural, environmental, developmental and social data to undertake studies with the ultimate goal of improving human health and well-being. Analyses of data from the recent Generation 1 year 26 follow-up are underway. ACTRN12617001599369.


Co‑cultivation of the anaerobic fungus Caecomyces churrovis with Methanobacterium bryantii enhances transcription of carbohydrate binding modules, dockerins, and pyruvate formate lyases on specific substrates.
Anaerobic fungi and methanogenic archaea are two classes of microorganisms found in the rumen microbiome that metabolically interact during lignocellulose breakdown. Here, stable synthetic co-cultures of the anaerobic fungus Caecomyces churrovis and the methanogen Methanobacterium bryantii (not native to the rumen) were formed, demonstrating that microbes from different environments can be paired based on metabolic ties. Transcriptional and metabolic changes induced by methanogen co-culture were evaluated in C. churrovis across a variety of substrates to identify mechanisms that impact biomass breakdown and sugar uptake. A high-quality genome of C. churrovis was obtained and annotated, which is the first sequenced genome of a non-rhizoid-forming anaerobic fungus. C. churrovis possess an abundance of CAZymes and carbohydrate binding modules and, in agreement with previous studies of early-diverging fungal lineages, N6-methyldeoxyadenine (6mA) was associated with transcriptionally active genes. Co-culture with the methanogen increased overall transcription of CAZymes, carbohydrate binding modules, and dockerin domains in co-cultures grown on both lignocellulose and cellulose and caused upregulation of genes coding associated enzymatic machinery including carbohydrate binding modules in family 18 and dockerin domains across multiple growth substrates relative to C. churrovis monoculture. Two other fungal strains grown on a reed canary grass substrate in co-culture with the same methanogen also exhibited high log2-fold change values for upregulation of genes encoding carbohydrate binding modules in families 1 and 18. Transcriptional upregulation indicated that co-culture of the C. churrovis strain with a methanogen may enhance pyruvate formate lyase (PFL) function for growth on xylan and fructose and production of bottleneck enzymes in sugar utilization pathways, further supporting the hypothesis that co-culture with a methanogen may enhance certain fungal metabolic functions. Upregulation of CBM18 may play a role in fungal-methanogen physical associations and fungal cell wall development and remodeling.


Selectivity in Infant Social Referencing.
In laboratory studies of social referencing, infants as young as 12 months have been reported to prefer looking at the experimenter over the caregiver for clarifying information. From an expertise perspective, such behavior could be interpreted as if the infant seeks information from others and can discriminate between persons who have or do not have relevant information to provide in the laboratory. If this is the case, higher order cognitive capacities might be involved in infant selectivity in looking in social referencing situations. However, it has also been proposed that associative learning processes might account for infant preferences in such studies. To examine whether an expertise perspective or if more basic learning processes best explain infant selectivity in looking, 40 12-month-old infants were assigned to 1 of 2 comparable conditions. The experimenter versus the caregiver presented an ambiguous toy and delivered positive information about the toy. The infants preferred to look at the experimenter and they regulated their behavior more in accordance with information coming from the experimenter. Thus, an associative learning account cannot explain infant preferences in looking. The results are discussed in terms of an expertise perspective.


Biochemical identification of a neutral sphingomyelinase 1 (NSM1)-like enzyme as the major NSM activity in the DT40 B-cell line: absence of a role in the apoptotic response to endoplasmic reticulum stress.
DT40 cells have approx. 10-fold higher Mg2+-dependent neutral sphingomyelinase (NSM) activity in comparison with other B-cell lines and contain very low acidic sphingomyelinase activity. Purification of this activity from DT40 cell membranes suggested the presence of one major NSM isoform. Although complete purification of this isoform could not be achieved, partially purified fractions were examined further with regard to the known characteristics of previously partially purified NSMs and the two cloned enzymes exhibiting in vitro NSM activity (NSM1 and NSM2). For a direct comparative study, highly purified brain preparations, purified NSM1 protein and Bacillus cereus enzyme were used. Analysis of the enzymic properties of the partially purified DT40 NSM, such as cation dependence, substrate specificity, redox regulation and stimulation by phosphatidylserine, together with the localization of this enzyme to the endoplasmic reticulum (ER), suggested that this NSM from DT40 cells corresponds to NSM1. Further studies aimed to correlate presence of the high levels of this NSM1-like activity in DT40 cells with the ability of these cells to accumulate ceramide and undergo apoptosis. When DT40 cells were stimulated to apoptose by a variety of agents, including the ER stress, an increase in endogenous ceramide levels was observed. However, these responses were not enhanced compared with another B-cell line (Nalm-6), characterized by low sphingomyelinase activity. In addition, DT40 cells were not more susceptible to ceramide accumulation and apoptosis when exposed to the ER stress compared with other apoptotic agents. Inhibition of de novo synthesis of ceramide partially inhibited its accumulation, indicating that the ceramide production in DT40 cells could be complex and, under some conditions, could involve both sphingomyelin hydrolysis and ceramide synthesis.


Chronic idiopathic effusive pericarditis in a patient with 'cor pulmonale'.
Chronic idiopathic pericarditis is a diagnosis of exclusion that is estimated to occur in 3.5% of primary pericardial disease. It is possible that many of these cases are secondary to unrecognised viral infections. The natural history is variable and treatment should be individualised, although chronic colchicines administration may hold promise as a newer therapeutic modality in symptomatic recurrent pericarditis. This case presentation illustrates the diagnostic and therapeutic difficulties that may arise in such patients.


Determinants of carriage of resistant Staphylococcus aureus among S. aureus carriers in the Indonesian population inside and outside hospitals.
To identify determinants of carriage of resistant Staphylococcus aureus in both hospitalized patients and individuals from the community in two urban centres in Indonesia. Staphylococcus aureus cultures and data on recent antibiotic use, demographic, socioeconomic, disease-related and healthcare-related variables were collected from 3995 community dwellers and hospitalized persons. Nasal S. aureus carriage was found in 362 persons (9.1%). Logistic regression analysis was performed to identify which variables were independently associated with carriage of resistant S. aureus. The penicillins were the most frequently used antibiotics both in the community and in hospitalized patients. In the community, admission to a hospital was associated with carriage of S. aureus resistant to any of the tested antibiotics [odds ratio (OR) 2.5, 95% confidence interval (95% CI) 1.3-4.9] and any tetracycline resistance (OR 2.4, 95% CI 1.1-5.1). Having no symptoms was associated with less carriage of S. aureus with resistance to any of the tested antibiotics (OR 0.5, 95% CI 0.3-0.9) and any tetracycline resistance (OR 0.5, 95% CI 0.3-0.9). Crowding (OR 4.5, 95% CI 1.2-4.9) and low income (OR 8.9, 95% CI 1.8-43.9) were associated with multidrug resistance. In hospitalized patients, the use of penicillins was associated with resistance to any of the tested antibiotics (OR 3.9, 95% CI 1.4-11.6) and any tetracycline resistance (OR 3.7, 95% CI 1.1-12.0). Antibiotic policies including proper diagnosis, treatment and drug delivery process should be made by healthcare providers in Indonesia to help limit the emergence of antibiotic resistance.


Changes in levels of catalase and glutathione in erythrocytes of patients with stable asthma, treated with beclomethasone dipropionate.
In asthmatic patients, antioxidant defence is decreased. Although inhaled corticosteroids decrease asthmatic inflammation and modulate reactive oxygen species (ROS) generation, little is known of their effect on cellular antioxidant levels. The aim of this study was to evaluate the effect of inhaled beclomethasone dipropionate (BDP; 1,000 microg x day(-1)) on erythrocyte antioxidant levels in stable asthmatic patients. Forty patients with stable, mild asthma were treated in a double-blind, placebo-controlled, parallel-group study with BDP 250 microg, two puffs b.i.d. for 6 weeks. At entry and every 2 weeks during treatment, erythrocyte antioxidant levels, haematological parameters, pulmonary function tests and asthma symptoms were determined. The results show that during treatment with BDP, erythrocyte catalase levels increased (at entry (mean +/-SEM) 41+/-4, after 6 weeks 54+/-4 micromol H2O2 x min(-1) x g haemoglobin (Hb)(-1), p = 0.05 in comparison with placebo). Erythrocyte total glutathione levels significantly decreased after 6 weeks treatment with BDP (from 7.0+/-0.4 to 6.6+/-0.3 micromol x g Hb(-1) (p = 0.04)). In the BDP-treated patients, blood eosinophil counts were higher in patients who responded with an increase in erythrocyte catalase levels during BDP treatment, as compared to those not responding ((mean +/-SEM) 340+/-39 and 153+/-52x10(6) cells x L(-1), respectively, p = 0.05). The present study shows that treatment with inhaled bedomethasone dipropionate results in changes in antioxidant levels in erythrocytes of patients with stable, mild asthma.


Biopersistence and brain translocation of aluminum adjuvants of vaccines.
Aluminum oxyhydroxide (alum) is a crystalline compound widely used as an immunological adjuvant of vaccines. Concerns linked to the use of alum particles emerged following recognition of their causative role in the so-called macrophagic myofasciitis (MMF) lesion detected in patients with myalgic encephalomyelitis/chronic fatigue/syndrome. MMF revealed an unexpectedly long-lasting biopersistence of alum within immune cells in presumably susceptible individuals, stressing the previous fundamental misconception of its biodisposition. We previously showed that poorly biodegradable aluminum-coated particles injected into muscle are promptly phagocytosed in muscle and the draining lymph nodes, and can disseminate within phagocytic cells throughout the body and slowly accumulate in brain. This strongly suggests that long-term adjuvant biopersistence within phagocytic cells is a prerequisite for slow brain translocation and delayed neurotoxicity. The understanding of basic mechanisms of particle biopersistence and brain translocation represents a major health challenge, since it could help to define susceptibility factors to develop chronic neurotoxic damage. Biopersistence of alum may be linked to its lysosome-destabilizing effect, which is likely due to direct crystal-induced rupture of phagolysosomal membranes. Macrophages that continuously perceive foreign particles in their cytosol will likely reiterate, with variable interindividual efficiency, a dedicated form of autophagy (xenophagy) until they dispose of alien materials. Successful compartmentalization of particles within double membrane autophagosomes and subsequent fusion with repaired and re-acidified lysosomes will expose alum to lysosomal acidic pH, the sole factor that can solubilize alum particles. Brain translocation of alum particles is linked to a Trojan horse mechanism previously described for infectious particles (HIV, HCV), that obeys to CCL2, signaling the major inflammatory monocyte chemoattractant.


Efficacy and safety of autologous bone marrow derived hematopoietic stem cell transplantation in patients with type 2 DM: A 15 months follow-up study.
there are dearths of studies describing the effect of autologous bone marrow derived stem cell transplantation (ABMSCT) through targeted approach in Type 2 Diabetes Mellitus. This study reports the efficacy and safety of super-selective injection of ABMSCT in T2DM. Ten patients (8 men and 2 women) with T2DM, with duration of disease >5 years and with documented triple drug failure receiving insulin (0.7 U/Kg/day), metformin and pioglitazone underwent super-selective injection of stem cells into superior pancreaticoduodenal artery under fluoroscopic guidance. The primary outcome measure was decrease in insulin requirement by ≥50% (defined as responders), while secondary endpoints were improvement in glucagon stimulated C-peptide levels, changes in weight, HbA1c, lipid profile and quality of life (QOL) at the end of 15 months. Six patients (60%) were 'responders' at 15 months of follow-up showing a reduction in mean insulin requirement by 74% as compared to baseline and one patient was off-insulin till the end of the study. Mean HbA1c reduction in 'responders' was 1.1% (8.1 ± 0.5% to 7.0 ± 0.6%, P = 0.03), accompanied with a significant improvement in glucagon stimulated C-peptide levels (P = 0.03), Homeostasis Model Assessment -β (P = 0.03) and QOL scores. However, 'non-responders' did not show any significant alterations in these parameters. No serious adverse events were noted. Our observations indicate that ABMSCT is effective in management of T2DM and its efficacy is maintained over a period of 15 months without any adverse events. However, more number of patients and longer duration of follow-up are required to substantiate these observations.


Analyses of the modulatory effects of antibacterial silver doped calcium phosphate-based ceramic nano-powder on proliferation, survival, and angiogenic capacity of different mammalian cells in vitro.
In this study, the antibacterial, cytotoxic, and angiogenic activities of silver doped calcium phosphate-based inorganic powder (ABT or PAG) were systematically investigated. ABT powders containing varying silver content were fabricated using a wet chemical manufacturing method. Antibacterial efficiencies of the ABT powders were investigated using a standard test with indicator bacteria and yeast. The cytotoxic effects of ABT on three different fibroblast cells and human umbilical vein endothelial cells (HUVECs) were assessed using MTT assay. ABT powder exhibits concentration-related cytotoxicity characteristics. Apoptotic activity, attachment capability, and wound healing effects were examined on fibroblasts. The angiogenic activity of ABT was investigated by tube formation assay in HUVECs; 10 μg ml(-1) and 100 μg ml(-1) concentrations of the highest metal ion content of ABT did not disrupt the tube formation of HUVECs. All these tests showed that ABT does not compromise the survival of the cells and might impose regeneration ability to various cell types. These results indicate that silver doped calcium phosphate-based inorganic powder with an optimal silver content has good potential for developing new biomaterials for implant applications.


Mental health prevalence and predictors among university students in nine countries during the COVID-19 pandemic: a cross-national study.
The student population has been highly vulnerable to the risk of mental health deterioration during the coronavirus disease (COVID-19) pandemic. This study aimed to reveal the prevalence and predictors of mental health among students in Poland, Slovenia, Czechia, Ukraine, Russia, Germany, Turkey, Israel, and Colombia in a socioeconomic context during the COVID-19 pandemic. The study was conducted among 2349 students (69% women) from May-July 2020. Data were collected by means of the Generalized Anxiety Disorder (GAD-7), Patient Health Questionnaire (PHQ-8), Perceived Stress Scale (PSS-10), Gender Inequality Index (GII), Standard & Poor's Global Ratings, the Oxford COVID-19 Government Response Tracker (OxCGRT), and a sociodemographic survey. Descriptive statistics and Bayesian multilevel skew-normal regression analyses were conducted. The prevalence of high stress, depression, and generalized anxiety symptoms in the total sample was 61.30%, 40.3%, and 30%, respectively. The multilevel Bayesian model showed that female sex was a credible predictor of PSS-10, GAD-7, and PHQ-8 scores. In addition, place of residence (town) and educational level (first-cycle studies) were risk factors for the PHQ-8. This study showed that mental health issues are alarming in the student population. Regular psychological support should be provided to students by universities.


Unmaking a murderer: behaviour sequence analysis of false confessions.
The causes of false confessions are an important issue in legal studies and forensics. Recent advances in DNA testing have increased the number of proven false confessions; however, there are probably many cases without scientific evidence to refute the guilty verdict. The current research provides a novel approach to understand the structure and process of police interrogation techniques, in the United States behaviour sequence analysis. This method allows complex interactions in interrogations to be broken down and the progression of techniques analysed clearly. A case study is provided of an individual who confessed to a series of very serious crimes. The results show that several psychological techniques, such as leading statements, pressure, empathy, and inducements, are used, in increasing frequency, which resulted in the suspect's confession. This research provides support for behaviour sequence analysis as a new method to understand the structure of police interrogations and how psychological techniques may be used to gain false confessions.


Intranasal immunization with C5a peptidase prevents nasopharyngeal colonization of mice by the group A Streptococcus.
Early inflammatory events are initiated by phased production of C5a and interleukin-8 in tissue. Most serotypes of group A streptococci express a surface-bound peptidase (SCPA) which specifically cleaves mouse and human C5a chemotaxins. This study investigates the impact of SCPA on colonization of the nasopharyngeal mucosa of mice and evaluates its potential to induce protective immunity. Two strains, serotypes M6 and M49, which contain insertion and deletion mutations in the SCPA gene (scpA) and represent the two major subdivisions of group A streptococci, were characterized and compared in a mouse intranasal infection model. In this model, SCPA mutants were more rapidly cleared from the nasopharynges of inoculated mice compared with wild-type strains. A 2,908-bp fragment of scpA49 gene, obtained by PCR, was ligated to the expression vector pGEX-4T-1 and expressed in Escherichia coli. The affinity-purified deltaSCPA49 protein proved to be highly immunogenic in mice and rabbits. Although the purified deltaSCPA49 immunogen lacked enzymatic activity, it induced high titers of rabbit antibodies which were able to neutralize peptidase activity associated with M1, M6, M12, and M49 streptococci in vitro. This result confirmed that antipeptidase antibodies lack serotype specificity. Intranasal immunization of mice with the deleted form of the SCPA49 protein stimulated significant levels of specific salivary secretory immunoglobulin A (IgA) and serum IgG antibodies and reduced the potential of wild-type M1, M2, M6, M11, and M49 streptococci to colonize. These experiments suggest a new approach to vaccine development for prevention of streptococcal pharyngitis.


Hand grip strength in the adult Malaysian population.
To measure the hand grip strength of Malaysians aged 18 to 65 years. Between January and April 2003, 412 subjects (200 women and 212 men) were recruited from staff, students, and visitors of the University of Malaya Medical Centre. Socioeconomic, general health, and lifestyle data were collected from each subject using a standard questionnaire. Weight and height were measured prior to testing. Standardised positioning and instructions based on several hand grip protocols were used. Data were collected using the LIDO kinetic work set. 93% of the subjects were right-hand dominant and 7% were left-hand dominant. Hand grip strength was significantly correlated with hand dominance, gender, occupation, height, and weight, but not body mass index. No significant differences in grip strength were noted with regard to race or level of income. Men were stronger than women in all age-groups, with a ratio of 1.75:1. In both right- and left-hand dominant groups, the dominant hand was consistently stronger than the non-dominant side, with a ratio of 1.12:1 in the right-hand dominant group and 1.05:1 in the left-hand dominant group. The strongest hand grip strength in the right-hand dominant group occurred in the age-group of 25 to 34 years; in the left-hand dominant group it was in the age-group of 18 to 24 years. In western populations, the mean grip strength can be as much as 1.5 times greater than in the Malaysian population. Data derived from western populations cannot be applied to a comparable Malaysian population. Gender, hand dominance, age, occupation, weight, and height must be considered when establishing normal values for grip strength.


The "ups and downs" of signaling cascades in addiction.
Drug addiction is a chronic disease characterized by compulsive drug use despite the severe negative consequences associated with it. Repeated exposure to drugs of abuse results in molecular adaptations in neuronal signaling pathways, which eventually manifest in the complex behavioral alterations that characterize addiction. These include tolerance, sensitization, dependence, drug craving, and relapse. In this Review, we focus on recent studies highlighting signaling cascades initiated by cocaine, as a representative of a drug of abuse with a defined site of action, and alcohol, as a drug with an undefined primary site of action. Specifically, we describe recent studies that emphasize the role of protein-protein interactions, phosphorylation, and compartmentalization in the molecular mechanisms that result in the cellular and behavioral adaptations that underlie addiction. Signaling cascades that contribute to addiction, as well as those that protect or delay the development of addiction, are presented.


Decrease in cell free DNA levels following participation in stress reduction techniques among women undergoing infertility treatment.
This pilot study examined differences in plasma cell free DNA (CFD) levels based on practice of stress reduction techniques among 14 women undergoing in vitro fertilization (IVF) and embryo transfer (ET) treatment who had higher than normal levels of plasma CFD before beginning IVF treatment. Wilcoxon nonparametric tests were used to examine the significance of the rate of decline in CFD levels between the time points in each of the groups. A paired sample t-test examined the changes in CFD levels among each participant in each of the groups separately. We found that women who engaged in these techniques had reduced plasma CFD, below what is considered elevated in comparison to those who did not practice. High plasma CFD levels have been associated with IVF failure. Stress reduction techniques may facilitate physiological changes that lead to the reduction of plasma CFD levels.


Isolation of a novel rat neural progenitor clone that expresses Dlx family transcription factors and gives rise to functional GABAergic neurons in culture.
Gamma-aminobutyric acid (GABA) ergic interneurons are lost in conditions including epilepsy and central nervous system injury, but there are few culture models available to study their function. Toward the goal of obtaining renewable sources of GABAergic neurons, we used the molecular profile of a functionally incomplete GABAergic precursor clone to screen 17 new clones isolated from GFP(+) rat E14.5 cortex and ganglionic eminence (GE) that were generated by viral introduction of v-myc. The clones grow as neurospheres in medium with FGF2, and after withdrawal of FGF2, they exhibit varying patterns of differentiation. Transcriptional profiling and quantitative reverse transcriptase polymerase chain reaction (RT-PCR) indicated that one clone (GE6) expresses high levels of mRNAs encoding Dlx1, 2, 5, and 6, glutamate decarboxylases, and presynaptic proteins including neuropeptide Y and somatostatin. Protein expression confirmed that GE6 is a progenitor with restricted differentiation giving rise mostly to neurons with GABAergic markers. In cocultures with hippocampal neurons, GE6 neurons became electrically excitable and received both inhibitory and excitatory synapses. After withdrawal of FGF2 in cultures of GE6 alone, neurons matured to express βIII-tubulin, and staining for synaptophysin and vesicular GABA transporter were robust after 1-2 weeks of differentiation. GE6 neurons also became electrically excitable and displayed synaptic activity, but synaptic currents were carried by chloride and were blocked by bicuculline. The results suggest that the GE6 clone, which is ventrally derived from the GE, resembles GABAergic interneuron progenitors that migrate into the developing forebrain. This is the first report of a relatively stable fetal clone that can be differentiated into GABAergic interneurons with functional synapses.


Decreased endothelial pump function with aging.
Endothelial function may be affected by the endothelial cell loss and increased variability in cell shape and size (polymegathism) that accompany normal aging. Endothelial function can be evaluated by monitoring corneal hydration recovery following hypoxic stress. The authors compared corneal recovery and endothelial morphology between a group of younger (mean = 26.7 yr) and older (means = 65.7 yr) subjects with normal corneas. Edema (60 microns) was induced with hydrogel lenses worn with the eyes closed. Following lens removal, the decrease in corneal thickness was monitored for 4 hr with one eye open while the contralateral eye remained closed. For both age groups, corneal recovery followed a non-linear time course. The open eye required 2.5 hr and 3.0 hr to return to baseline for the younger and older age groups, respectively. Recovery during eye closure took 3.5 hr to reach the normal closed eye level for the younger subjects and was not complete at 4 hr for the older subjects. Recovery rates were significantly slower for the older vs younger subjects during the first 2 hr of closed eye recovery, 10.5 vs 15.0 microns/hr, and for the initial 1 hr of open eye recovery, 26.5 vs 35.6 microns/hr. For both age groups combined, the rate of recovery was negatively correlated with the coefficient of variation in cell area, r = -0.62 and -0.69 (P less than 0.01), for both closed and open eye recovery, respectively. When each morphological characteristic was isolated, the only significant correlation found was between the coefficient of variation in cell area and the rate of recovery during eye closure, r = -0.66 (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Detection of peste des petits ruminants virus in formalin-fixed tissues.
Peste des petits ruminants virus that causes a highly infectious and often fatal disease of sheep and goats is confirmed by various diagnostic techniques among them being isolation of the virus from cell culture systems, viral ribonucleic acid (RNA) detection by molecular assays, and viral antigen detection by immunocapture enzyme-linked immunosorbent assay (IC ELISA), immunohistochemistry (IHC), and AGAR gel test. Whereas most of the confirmatory diagnostic procedures require pathological samples to be stored frozen to preserve integrity of the peste des petits ruminants (PPR) virus RNA, samples for IHC tests are preserved in 10% formalin. In this study, nine formalin-fixed pathological samples from three goats suspected of PPR were processed for extraction of PPR viral RNA and analyzed for detection with real-time reverse transcription-polymerase chain reaction (qRT-PCR) assay. The results showed that five out of the nine tested samples returned positive for presences PPR viral genome. This study has established that field pathological samples of PPR-suspected cases, collected and stored in 10% formalin for up 2 years, could be used for PPR virus RNA extraction for disease virus confirmation.


mdm-2 expression correlates with wild-type p53 status in esophageal adenocarcinoma.
Several immunohistochemical studies showed that p53 protein is expressed in 50 to 80% of esophageal adenocarcinomas (EAs). Mutations of this tumor suppressor gene are present in 40 to 70% of EAs, so it is possible that p53 expression might occur as a result of mechanisms other than gene mutation. The human homologue of the murine double minute-2 gene (mdm-2) is a known regulator of p53 activity, and its expression results in stabilization of the wild-type p53 protein and loss of its tumor suppressor function. In this study, we evaluated the frequency of mdm-2 amplification and expression in EA and investigated the relationship between mdm-2 expression and p53 mutation. Thirty-three resection specimens of EAs and associated Barrett's esophagus were evaluated by immunohistochemical methods for p53 and mdm-2 expression. Sixteen of these cases were also evaluated for p53 mutations with use of polymerase chain reaction, single-strand conformational polymorphism, and DNA sequencing and for mdm-2 amplification with a differential polymerase chain reaction-based amplification analysis. Overexpression of p53 was present in 23 EAs (70%), and 18 EAs (55%) overexpressed mdm-2. p53 mutation was observed in 7 (43%) of 16 cases, whereas mdm-2 gene amplification was not detected in any. To summarize, we found substantial discordance of p53 immunohistochemical features and mutation in EA. Significant expression of mdm-2 occurred only in cases with wild-type p53, whereas all of the cases with p53 mutation showed little if any expression of mdm-2. Also, mdm-2 expression in cases with p53 overexpression but without p53 mutation exceeded mdm-2 expression in cases with p53 overexpression and p53 gene mutation. In cases without p53 mutation, overexpression of mdm-2 occurred in 50% of cases and might be responsible for stabilization of p53 protein and possible loss of tumor suppressor function.


Structure-activity relationships in the cytoprotective effect of caffeic acid phenethyl ester (CAPE) and fluorinated derivatives: effects on heme oxygenase-1 induction and antioxidant activities.
To determine the relationship between catechol ring modifications and the activity of caffeic acid phenethyl ester (CAPE) as a cytoprotective agent, six catechol ring-fluorinated CAPE derivatives were evaluated for their cytoprotective abilities, as well as for their antioxidant and heme oxygenase-1 (HO-1) inducing capacity in a human umbilical vein endothelial cell (HUVEC) model of oxidant stress. To ascertain the involvement of HO-1 induction in the cytoprotective effects of CAPE analogues, their ability to induce HO-1 at 20microM was determined by reverse transcriptase polymerase chain reaction, western blotting and the use of HO-1 inhibitor tin protoporphyrin IX. There was significant induction of HO-1 by CAPE derivatives. Inhibition of HO-1 enzymatic activity resulted in reduced cytoprotection. Modification of the catechol ring of CAPE by introduction of fluorine at various positions resulted in dramatic changes in cytoprotective activity. The maintenance of at least one hydroxyl group on the CAPE catechol ring and the phenethyl ester portion was required for HO-1 induction. CAPE and its derivatives were screened for their ability to scavenge intracellular reactive oxygen species generated in HUVECs by measuring 5-(and-6)-chlormethyl-2', 7'-dichlorodihydrofluorescein diacetate oxidation. The maintenance of 3, 4-dihydroxyl groups on the catechol ring was required for antioxidant activity, but antioxidant activity did not guarantee cytoprotection. Methylation or replacement of one hydroxyl group on the catechol ring of CAPE, however, provided both pro-oxidant and cytoprotective activities. These results indicate that the induction of HO-1 plays a more important role in the cytoprotective activity of CAPE derivatives than their direct antioxidant activity.


Luteolin alleviates LPS-induced bronchopneumonia injury in vitro and in vivo by down-regulating microRNA-132 expression.
Bronchopneumonia is a common multiple infection disease under 2 years old. Luteolin is a natural flavonoid widely distributed in plants with anti-inflammatory effect. This study aimed to explore the effects of luteolin on lipopolysaccharide (LPS)-induced bronchopneumonia injury in vitro and in vivo. Firstly, the viability and apoptosis of human bronchial epithelial BEAS-2B cells after luteolin treatment were assessed. Then, cells were treated with 10 μM LPS to simulate inflammatory injury. The potential protective effects of luteolin on LPS-induced BEAS-2B cell inflammatory injury were detected. Moreover, after LPS and/or luteolin treatment, the expression of microRNA-132 (miR-132) in BEAS-2B cells was measured. The roles of miR-132 in protective activity of luteolin were investigated. Finally, the LPS-induced bronchopneumonia murine model was established and the anti-inflammatory effects of luteolin in vivo were analyzed. The results showed that LPS decreased BEAS-2B cell viability, increased cell apoptosis and enhanced inflammatory cytokines expression. Luteolin alleviated the LPS-induced viability loss, apoptosis and elevated expression of inflammatory cytokines in a dose-dependent manner. Moreover, luteolin alleviated the LPS-induced miR-132 expression increase in BEAS-2B cells. Overexpression of miR-132 reversed the protective effects of luteolin on LPS-induced inflammatory injury. Mechanistically, luteolin mitigated LPS-induced activation of NF-κB signaling pathway by down-regulation of miR-132. Furthermore, we also found that luteolin alleviated LPS-induced bronchopneumonia model in vivo. In conclusion, this study revealed that luteolin alleviated LPS-induced bronchopneumonia injury in vitro and in vivo through down-regulating miR-132. These findings provide theoretical basis for deeply exploring the treatment of bronchopneumonia in children by using luteolin.


Piecewise synonyms for enhanced UMLS source terminology integration.
The UMLS contains more than 100 source vocabularies and is growing via the integration of others. When integrating a new source, the source terms already in the UMLS must first be found. The easiest approach to this is simple string matching. However, string matching usually does not find all concepts that should be found. A new methodology, based on the notion of piecewise synonyms, for enhancing the process of concept discovery in the UMLS is presented. This methodology is supported by first creating a general synonym dictionary based on the UMLS. Each multi-word source term is decomposed into its component words, allowing for the generation of separate synonyms for each word from the general synonym dictionary. The recombination of these synonyms into new terms creates an expanded pool of matching candidates for terms from the source. The methodology is demonstrated with respect to an existing UMLS source. It shows a 34% improvement over simple string matching.


Modulation of antigen processing by bound antibodies can boost or suppress class II major histocompatibility complex presentation of different T cell determinants.
Bound antibodies can modulate antigen processing but it is not clear to what extent this affects antigen presentation. Here we show that presentation of T cell determinants in tetanus toxin can be either enhanced or suppressed as a direct consequence of antibody modulation of antigen processing in human B lymphoblastoid cells. Remarkably, a single bound antibody or its Fab fragment can simultaneously enhance the presentation of one T cell determinant by more than 10-fold while strongly suppressing the presentation of a different T cell determinant. Biochemical analysis demonstrates that both the suppressed and boosted determinants fall within an extended domain of antigen stabilized or "footprinted" by this antibody during proteolysis. These results demonstrate that bound antibodies can modulate the capture of peptides by class II major histocompatibility complex (MHC), thus manipulating the T cell response towards or away from particular determinants. Altered processing of protein-protein complexes leading to enhanced loading of class II MHC and substantially lowered threshold for T cell activation suggests a novel mechanism that might reveal "cryptic" self determinants.


Human skeletal muscle cytosols are refractory to cytochrome c-dependent activation of type-II caspases and lack APAF-1.
Apoptotic regulatory mechanisms in skeletal muscle have not been revealed. This is despite indications that remnant apoptotic events are detected following exercise, muscle injury and the progression of dystrophinopathies. The recent elicitation of a cytochrome c-mediated induction of caspases has led to speculation regarding a cytochrome c mechanism in muscle. We demonstrate that cytosols from skeletal muscle biopsies from healthy human volunteers lack the ability to activate type-II caspases by a cytochrome c-mediated pathway despite the confirmed presence of both procaspase-3 and -9. This was not due to the presence of an endogenous inhibitor, as the muscle cytosols enhanced caspase activity when added to a control cytosol, subsequently activated by cytochrome c and dATP. In addition, we demonstrate that muscle cytosols lack the apoptosis protease activator protein-1 (APAF-1), both at the protein and mRNA levels. These data indicate that human skeletal muscle cells will be refractory to mitochondrial-mediated events leading to apoptosis and thus can escape a major pro-apoptotic regulatory mechanism. This may reflect an evolutionary adaptation of cell survival in the presence of the profusion of mitochondria required for energy generation in motility.


A fast, digitally controlled flow proportional gas injection system for studies in lung function.
The aim of this paper is to describe a device for flow proportional injection of tracer gas in the lungs of mechanically ventilated patients. This device may then be used for the study of the multiple breath indicator gas washout technique to determine the end-expiratory lung volume. Such a tracer gas injection device may also be used in the study of other techniques that rely on uptake and elimination of tracer gas by the lungs. In this paper, an injector is described which enables injection of indicator gas at a predetermined concentration in a breathing circuit independent of the type of breathing. The presented setup uses a control computer to produce steering signals to a multivalve array in proportion to the input breathing signals. The multivalve array consists of ten circular valves, each with a different diameter, which can be opened or closed individually according to the input signal of the array. By opening of a certain combination of valves an amount of sulphur hexafluoride gas proportional to the inspiratory breathing signal is released. The rate of transmission between the components of the injection system was 80 Hz. The injector has a full flow range between 0-10 L/min. The delay time between the breathing signal and the flow response was 70 ms. The aimed washin gas concentration of 1% SF6 was achieved after 0.5 s. The study describes the results of tests to determine valve-flow ratios, step response and dynamic response of the injector. The flow output response of the injector system was shown to increase in input frequencies above 3 Hz. The valve flow ratios showed the largest relative deviation in the two smallest valves of the 10 valve array, respectively 0.005 L/min (25%) and 0.002 L/min (20%). We conclude that the injector can achieve a stable concentration of indicator gas in a breathing system with an accuracy of 0.005 L/min to execute the multiple breath indicator washout test in human subjects. The results of the study indicate that the injector may be of use in other application fields in respiratory physiology in which breathing circuit injection of indicator gas is required.


Readmissions of medical patients: an external validation of two existing prediction scores.
Hospital readmissions are increasingly used as a quality indicator with a belief that they are a marker of poor care and have led to financial penalties in UK and USA. Risk scoring systems, such as LACE and HOSPITAL, have been proposed as tools for identifying patients at high risk of readmission but have not been validated in international populations. To perform an external independent validation of the HOSPITAL and LACE scores. An unplanned secondary cohort study. Patients admitted to the medical admission unit at the Hospital of South West Jutland (10/2008-2/2009; 2/2010-5/2010) and the Odense University Hospital (6/2009-8/2011) were analysed. Validation of the scores using 30 day readmissions as the endpoint was performed. A total of 19 277 patients fulfilled the inclusion criteria. Median age was 67 (range 18-107) years and 8977 (46.6%) were female. The LACE score had a discriminatory power of 0.648 with poor calibration and the HOSPITAL score had a discriminatory power of 0.661 with poor calibration. The HOSPITAL score was significantly better than the LACE score for identifying patients at risk of 30 day readmission (P < 0.001). The discriminatory power of both scores decreased with increasing age. Readmissions are a complex phenomenon with not only medical conditions contributing but also system, cultural and environmental factors exerting a significant influence. It is possible that the heterogeneity of the population and health care systems may prohibit the creation of a simple prediction tool that can be used internationally.


[Liver changes in workers at an oil refinery and in a reference population in the state of Bahia, Brazil].
To determine the prevalence of liver changes in workers at an oil refinery located in the state of Bahia, Brazil, as compared to a reference population with no occupational exposure to chemical products, and to describe the factors associated with the observed differences in prevalence. We studied workers at the refinery and at the company's central management office located in the city of Salvador, which is the state capital. Blood samples of 692 refinery workers and 377 workers from the reference population were analyzed. Cases were defined as individuals presenting high serum levels of both gammaglutamyltransferase (GGT) and alanine aminotransferase (ALT) (GGT > 50 U/L for males and > 32 U/L for females; ALT > 50 U/L). Data were collected on age, sex, weight, height, years working for the company, use of alcohol, smoking habits, physical exercise, occupational exposure to chemical products, use of personal safety equipment, and medical history of hepatitis, jaundice, and obesity. The prevalence of liver changes among refinery workers was 15.3% (95% CI: 12.5 to 18.1), vs. 3.8% (95% CI: 1,8 to 5,8) in the reference population. According to multiple logistic regression analysis, the prevalence of liver changes among refinery workers was 3.56 times greater (95% CI: 1,99 to 6,38) than in the reference population, regardless of the presence of other relevant co-variables, such as obesity, exercise, smoking, and alcohol consumption. These results suggest that occupational exposure may play a major role in the development of liver changes among workers at the oil refinery where the study was carried out.


High prevalence of tuberculosis and insufficient case detection in two communities in the Western Cape, South Africa.
In South Africa the estimated incidence of all forms of tuberculosis (TB) for 2008 was 960/100000. It was reported that all South Africans lived in districts with Directly Observed Therapy, Short-course. However, the 2011 WHO report indicated South Africa as the only country in the world where the TB incidence is still rising. To report the results of a TB prevalence survey and to determine the speed of TB case detection in the study communities. In 2005 a TB prevalence survey was done to inform the sample size calculation for the ZAMSTAR (Zambia South Africa TB and AIDS Reduction) trial. It was a cluster survey with clustering by enumeration area; all households were visited within enumeration areas and informed consent obtained from eligible adults. A questionnaire was completed and a sputum sample collected from each adult. Samples were inoculated on both liquid mycobacterium growth indicator tube (MGIT) and Löwenstein-Jensen media. A follow-up HIV prevalence survey was done in 2007. In Community A, the adjusted prevalence of culture positive TB was 32/1000 (95%CI 25-41/1000) and of smear positive TB 8/1000 (95%CI 5-13/1000). In Community B, the adjusted prevalence of culture positive TB was 24/1000 (95%CI 17-32/1000) and of smear positive TB 9/1000 (95%CI 6-15/1000). In Community A the patient diagnostic rate was 0.38/person-year while in community B it was 0.30/person-year. In both communities the adjusted HIV prevalence was 25% (19-30%). In both communities a higher TB prevalence than national estimates and a low patient diagnostic rate was calculated, suggesting that cases are not detected at a sufficient rate to interrupt transmission. These findings may contribute to the rising TB incidence in South Africa. The TB epidemic should therefore be addressed rapidly and effectively, especially in the presence of the concurrently high HIV prevalence.


Prior gold therapy does not influence the adverse effects of D-penicillamine in rheumatoid arthritis.
One hundred fourteen patients with definite or classic rheumatoid arthritis were followed prospectively between January 1976 and April 1981 to monitor their toxicity pattern to D-penicillamine. The influence of previous sodium aurothiomalate therapy on the toxicity pattern of D-penicillamine is described. There was no significant difference in overall outcome of the patients treated with D-penicillamine with respect to adverse effects, whether they had previous gold toxicity, previous gold therapy but no toxicity, or no previous gold therapy. The time from gold toxicity to the start of D-penicillamine therapy was greater in those who did not develop D-penicillamine toxicity compared with those who did. This difference just reached statistical significance. Total gold salt received had no effect on eventual outcome of D-penicillamine treatment, and the toxicity pattern of D-penicillamine in those patients who had previous gold therapy was similar to those patients who had never received gold therapy.


Comparative Efficacy of Injection Triamcinolone Acetonide given Intralesionally and through Microneedling in Alopecia areata.
Alopecia areata (AA) manifests as patchy hair loss and intralesional corticosteroid (ILCS) is usual therapeutic choice in limited disease. Microneedling is used for uniform delivery of topical agent to relatively larger areas may prove to be more efficacious than traditional ILCS. The present study prospectively compared microneedling to traditional intralesional delivery of triamcinolone acetonide (TA). Prospective randomized comparative study in 60 patients of AA restricted to scalp not requiring systemic treatment randomly divided into two equal groups. Group 1 patients underwent microneedling with local application of injectable TA and Group 2 patients were given injectable TA intradermally for a total of three sessions at 3 weeks interval. A mean regrowth of 66.36% in Group 1 and 69.75% in Group 2 at week 9 was seen which was comparative with no significant statistical difference between the two groups (P = 0.664). Thirteen patients achieved 100% regrowth at week 9 in Group 1 and 16 patients achieved 50%-99% regrowth in Group 2. ILCSs have been cornerstone in the treatment of limited AA, but depth of injecting drug cannot be controlled, microneedling whereas is an effective drug delivery system and also causes release of growth factors. In this study, injectable TA used intralesionally and topically with microneedling had nearly similar efficacy in causing regrowth of hair with microneedling resulting in a more uniform but less dense regrowth of hair with lesser adverse effects.


Social distancing enhanced automated optimal design of physical spaces in the wake of the COVID-19 pandemic.
As the COVID-19 pandemic unfolds, manually enhanced ad-hoc solutions have helped the physical space designers and decision makers to cope with the dynamic nature of space planning. Due to the unpredictable nature by which the pandemic is unfolding, the standard operating procedures also change, and the protocols for physical interaction require continuous reconsideration. Consequently, the development of an appropriate technological solution to address the current challenge of reconfiguring common physical environments with prescribed physical distancing measures is much needed. To do this, we propose a design optimization methodology which takes the dimensions, as well as the constraints and other necessary requirements of a given physical space to yield optimal redesign solutions on the go. The methodology we propose here utilizes the solution to the well-known mathematical circle packing problem, which we define as a constrained mathematical optimization problem. The resulting optimization problem is solved subject to a given set of parameters and constraints - corresponding to the requirements on the social distancing criteria between people and the imposed constraints on the physical spaces such as the position of doors, windows, walkways and the variables related to the indoor airflow pattern. Thus, given the dimensions of a physical space and other essential requirements, the solution resulting from the automated optimization algorithm can suggest an optimal set of redesign solutions from which a user can pick the most feasible option. We demonstrate our automated optimal design methodology by way of a number of practical examples, and we discuss how this framework can be further taken forward as a design platform that can be implemented practically.


Restoration of intracellular ATP production in banked red blood cells improves inducible ATP export and suppresses RBC-endothelial adhesion.
Transfusion of banked red blood cells (RBCs) has been associated with poor cardiovascular outcomes. Storage-induced alterations in RBC glycolytic flux, attenuated ATP export, and microvascular adhesion of transfused RBCs in vivo could contribute, but the underlying mechanisms have not been tested. We tested the novel hypothesis that improving deoxygenation-induced metabolic flux and the associated intracellular ATP generation in stored RBCs (sRBCs) results in an increased extracellular ATP export and suppresses microvascular adhesion of RBCs to endothelium in vivo following transfusion. We show deficient intracellular ATP production and ATP export by human sRBCs during deoxygenation (impairments ~42% and 49%, respectively). sRBC pretreatment with a solution containing glycolytic intermediate/purine/phosphate precursors (i.e., "PIPA") restored deoxygenation-induced intracellular ATP production and promoted extracellular ATP export (improvement ~120% and 50%, respectively). In a nude mouse model of transfusion, adhesion of human RBCs to the microvasculature in vivo was examined. Only 2% of fresh RBCs (fRBCs) transfused adhered to the vascular wall, compared with 16% of sRBCs transfused. PIPA pretreatment of sRBCs significantly reduced adhesion to just 5%. In hypoxia, adhesion of sRBCs transfused was significantly augmented (up to 21%), but not following transfusion of fRBCs or PIPA-treated sRBCs (3.5% or 6%). Enhancing the capacity for deoxygenation-induced glycolytic flux within sRBCs increases their ability to generate intracellular ATP, improves the inducible export of extracellular anti-adhesive ATP, and consequently suppresses adhesion of stored, transfused RBCs to the vascular wall in vivo.


Cardiovascular involvement in osteogenesis imperfecta.
While aortic root dilatation and valvular dysfunction have been well-documented in osteogenesis imperfecta (OI), the nature and extent of cardiovascular involvement in OI have not been clearly delineated. A clinical and echocardiographic survey involving 109 individuals with various nonlethal OI syndromes from 66 separate families was undertaken. Clinically discernible valvular dysfunction was encountered in only four of the 109 individuals (aortic regurgitation in two, aortic stenosis in one, and mitral valve prolapse in one), none of whom were related. Aortic root dilatation was recognized echocardiographically in eight (12.1%) of 66 individuals comprising a subset of the sample in which each family was represented by a single individual. The extent of the aortic root dilatation was mild (the largest dimension measuring 4.3 cm) and was unrelated to the age of the individual. Dilatation was seen in each of the different OI syndromes but was strikingly segregated within certain families (p less than .001). In the same subset of 66 individuals, mitral valve prolapse was encountered in two or 6.9% of the 29 individuals aged 15 years or greater in whom adequate studies were obtained. This observed frequency was not different from that seen in a normal adult population. Aortic root dilatation appears to represent a distinct phenotypic trait in patients with OI that is nonprogressive and occurs in about 12% of affected individuals. Whether mitral valve prolapse should be considered as a part of the cardiovascular phenotype in OI, or alternately segregates as an independent autosomal dominant trait has yet to be determined.


Flexibility and posture assessment in relation to hamstring injury.
Posture and flexibility were assessed in 34 athletes. Subjects were divided into two groups: (1) a noninjured group that did not have a history of hamstring strain injury within the previous 12 months; (2) an injured group that had a history of hamstring strain within the previous 12 months. Ten postural components were assessed: head erectness; shoulder symmetry; spinal curvature; hip symmetry; foot and ankle alignment; knee hyperextension; upper back roundness; trunk erectness; abdomen protrusion; and lumbar lordosis. Hamstring flexibility was assessed in both legs. Results indicated no difference (P > 0.05) in flexibility between groups (mean(s.d.) of both legs was: noninjured = 77.1(9.3) degrees, injured = 77.8(9.2) degrees. Also no difference (P > 0.05) was observed between the injured limb and the noninjured limb for injured subjects (injured limb = 78.1(11.1) degrees, noninjured limb = 77.5(8.1) degrees. A significant difference (P < 0.01) between groups occurred in low back posture (lumbar lordosis). No other difference occurred in the remaining nine posture components between groups. Intercorrelation coefficients among posture components indicated that at best only 53% of common variance existed between any two components (head and shoulder components: r = 0.73, P < 0.01). All other correlations indicated less than 40% common variance between components. The results of the study indicate that while differences in hamstring flexibility are not evident between injured and noninjured groups poorer low back posture was found in the injured group. Regular monitoring of posture in athletes is recommended.


Effective date of requirement for premarket approval for cardiovascular permanent pacemaker electrode. Final rule.
The Food and Drug Administration (FDA) is issuing a final rule to require the filing of a premarket approval application (PMA) or a notice of completion of a product development protocol (PDP) for the cardiovascular permanent pacemaker electrode. The Agency has summarized its findings regarding the degree of risk of illness or injury designed to be eliminated or reduced by requiring the device to meet the statute's approval requirements and the benefits to the public from the use of the device. This action implements certain statutory requirements.


Pathogenesis of Lassa fever.
Lassa virus, an Old World arenavirus (family Arenaviridae), is the etiological agent of Lassa fever, a severe human disease that is reported in more than 100,000 patients annually in the endemic regions of West Africa with mortality rates for hospitalized patients varying between 5-10%. Currently, there are no approved vaccines against Lassa fever for use in humans. Here, we review the published literature on the life cycle of Lassa virus with the specific focus put on Lassa fever pathogenesis in humans and relevant animal models. Advancing knowledge significantly improves our understanding of Lassa virus biology, as well as of the mechanisms that allow the virus to evade the host's immune system. However, further investigations are required in order to design improved diagnostic tools, an effective vaccine, and therapeutic agents.


Dissection of the xeroderma pigmentosum group C protein function by site-directed mutagenesis.
Xeroderma pigmentosum group C (XPC) protein is a sensor of helix-distorting DNA lesions, the function of which is to trigger the global genome repair (GGR) pathway. Previous studies demonstrated that XPC protein operates by detecting the single-stranded character of non-hydrogen-bonded bases opposing lesion sites. This mode of action is supported by structural analyses of the yeast Rad4 homologue that identified critical side chains making close contacts with a pair of extrahelical nucleotides. Here, alanine substitutions of the respective conserved residues (N754, F756, F797, F799) in human XPC were tested for DNA-binding activity, accumulation in tracks and foci of DNA lesions, nuclear protein mobility, and the induction of downstream GGR reactions. This study discloses a dynamic interplay between XPC protein and DNA, whereby the association with one displaced nucleotide in the undamaged strand mediates the initial encounter with lesion sites. The additional flipping-out of an adjacent nucleotide is necessary to hand over the damaged site to the next GGR player. Surprisingly, this mutagenesis analysis also reveals that the rapid intranuclear trafficking of XPC protein depends on constitutive interactions with native DNA, implying that the search for base damage takes place in living cells by a facilitated diffusion process.