Molecular surveillance of antimalarial partner drug resistance in sub-Saharan Africa: a spatial-temporal evidence mapping study.
Molecular markers for antimalarial drug resistance can be used to rapidly monitor the emergence and spatial distribution of resistance to artemisinin-based combination therapies (ACTs). Little has been done to analyse molecular surveillance efforts or to assess surveillance coverage. This study aimed to develop an evidence map to characterise the spatial-temporal distribution and sampling methodologies of drug resistance surveillance in sub-Saharan Africa, specifically focusing on markers associated with ACT partner drugs. By use of a systematic search, we identified studies that reported data on the following mutations associated with ACT partner drug resistance: pfmdr1 Asn86Tyr, Tyr184Phe, Asp1246Tyr, and copy number variation and pfcrt Lys76Thr, with sample collection occurring in sub-Saharan Africa between Jan 1, 2004, and Dec 31, 2018, corresponding to the uptake of ACTs. For each identified study, we extracted information on its sampling and laboratory methods, author and publication affiliations, years of sampling and of publication, geographic coordinates of the study sites, and prevalence of the partner drug resistance-associated markers. We used linear models to test whether urbanicity, population density, and endemicity were predictors of drug resistance survey sites and linear regressions to identify associations between the number of resistance surveys within a given country and the at-risk malaria population in 2010, the per-capita GDP in 2010, and the mean amount of funding directed to malaria and to determine trends in marker prevalence over time. For country case studies with three or more datapoints, we assessed global spatial autocorrelation using Moran's I. Our search yielded 254 studies encompassing 492 year-specific and location-specific surveys from 35 malaria-endemic countries, the most complete set of molecular partner drug surveillance data to date. We observed a median time lag of 3·1 years (95% CI 1·0-7·7) from final sample acquisition to publication. 22 (49%) of the 44 countries in the study region conducted, on average, one or fewer studies every 3 years. The locations of surveillance sites were positively associated with urbanicity (p<0·0001), and the abundance of country-level data was associated with reported donor funding in 2004-18 (p=0·0011) and local government funding in 2004-09 (p=0·014). Nearly all molecular markers displayed significant regional trends over time and global spatial autocorrelation in space. For selected countries with more widespread coverage of surveillance data, some markers also displayed spatial heterogeneity. In most sub-Saharan countries, molecular data on antimalarial resistance might not be representative of the temporal and geographic heterogeneity of partner drug resistance, and likely do not represent the true spatially dependent distribution of partner drug resistance. Our results highlight several inefficiencies that can be improved upon to develop more accurate data landscapes, including the expansion of sentinel surveillance systems, syndemic usage of research samples, and increased participation in reporting published and unpublished data to centralised platforms.


Marginal Traction Alopecia: Hair Care Practices, Severity Score and Trichoscopic Features in Lagos, Nigeria.
Marginal traction alopecia (MTA) is commonly observed in adult females. Studies of prevalence, severity, trichoscopy and severity scoring are few. The aim of this study was to determine the community prevalence, pattern, severity score and trichoscopy features of MTA amongst adult Nigerian females. Patient and method: This was a cross-sectional descriptive study of a cohort of 207 adult females in a community in Lagos, Nigeria. The participants were clinically evaluated for MTA. Sociodemographic, clinical and trichoscopy data was documented. Severity score was documented using the MTA severity score. Data was analyzed using SPSS version 22. The mean age of participants was 43±12.8 years and the prevalence of MTA was 76.3%. Marginal traction alopecia was frontoccipital in 65.2%. The MTA score was 1-3 in 22.2%, 4-6 in 29.7% and ≥6 in 48.1%. Fringe sign was observed in 30.4%, folliculitis in 9.5% and 13.9% had tight weaves. Trichoscopic features included preserved honeycomb pigment, preserved white dot architecture, varying hair length and single hairs from hair follicles. Marginal traction alopecia is common in Nigerian women. It is mostly severe, severity is not significantly associated with haircare practices and it is represented by common trichoscopy features. Appropriate awareness and education of females on hair styles can mitigate this hair loss.


In vivo transfection of the mouse vas deferens.
To explore the possibility that specific characteristics of the epithelium of the male tract can be modified, transfections of the mouse vas deferens have been performed using in vivo injections of cationic DNA/liposome complexes. Gene transfer was done employing the reporter genes pEGFP-C1 encoding Green Fluorescent Protein (GFP) and pCMV-nls-beta encoding the nuclear beta-Galactosidase (beta-Gal). Foreign gene expression reached a maximum of 6.8% in the epithelial cells of the vas after treatment with the nuclear beta-Gal gene construction and of 13.3% after employing the GFP gene construction. Expression of the GFP gene appeared from one week up to three months following injection, and it appeared as patches of modified cells along the epithelium. Results from immunocytochemistry and Western Blotting support the conclusion that transfection of epithelial cells was achieved. We have also transfected the vas using gene constructions that express secretory proteins--specifically, the reporter system pSEAP-control that expresses a secretory form of human placental alkaline phosphatase, and the pGFP-Ctk-37 that expresses a secretion form of GFP. In both cases, the fluids expressed from the transfected vas showed a significant increase of alkaline phosphatase activity after pSEAP transfection and the presence of GFP protein when pGFP-Ctk-37 gene construction was employed. Our results indicate that the vas can be transfected in vivo using liposomes as vectors of foreign genes and that the vas fluid contents can be modified.


Torsades de pointes: unanswered questions.
Torsade de pointes (Tdp) is a polymorphic ventricular tachycardia (VT) in which the axis of the QRS complex changes direction after a certain number of complexes as if the complex rotated around the baseline. Tdp is usually associated with QT prolongation, and dispersion of ventricular repolarlization (DR). Experimental models of tdp are usually associated with induction of early after depolarizations (EADs). Several aspects of the pathogenesis of tdp are incompletely understood. The purpose of this article is to propose the directions in research that may increase our current understanding of the factors responsible for tdp. The most plausible hypotheses requiring further supporting evidence are: 1. The occurrence of tdp requires the presence of DR i.e. tdp does not occur in the absence of DR. 2. EADs appear to play an important role as a trigger to tdp in the animal models, but more evidence are needed at the clinical level. 3. EADs may be responsible for arrhythmias other than tdp. 4. The greater incidence of tdp in the females than in tha males may be attributed to differences in the duration of the QT interval and different morphology of the ST-segment and the T-wave. The above gender differences may be caused by the effects of gonadal hormones which modulate some of the membrane currents flowing during early ventricular repolarization.


Women with primary antibody deficiencies requiring IgG replacement therapy: their perception of prenatal care during pregnancy.
To investigate how a group of women with primary antibody deficiencies (PAD) and receiving replacement therapy with IgG experienced the care they received in their prenatal clinics in relation to PAD and IgG therapy. An exploratory study using a written questionnaire. The study originates from an immunodeficiency unit but evaluates care experienced at prenatal clinics. Nine women (25-43 years) attending an immunodeficiency unit and who fulfilled inclusion criteria for simultaneously having PAD, replacement IgG therapy, and full-term pregnancy (the latter within the past 5 years). Women's perception of the response of midwives and physicians at their prenatal clinics to their PAD and IgG therapy during pregnancy. Women perceived that the obstetricians and the midwives had insufficient knowledge about PAD and IgG replacement therapy. Two women reported that their IgG therapy during pregnancy had been questioned. All nine women felt marginalized and unheard by staff regarding their PAD and need for IgG therapy. However, the women were satisfied with the checkups regarding the pregnancy as such. This study is the first attempt to investigate the prenatal experience of women with PAD (Search of PubMed, 1980 to present, including search terms primary immunodeficiency, pregnancy, and prenatal care). This study demonstrates that increased knowledge about PAD and IgG replacement therapy among midwives and physicians working in prenatal care clinics is needed. This can prevent misleading advice that puts the health of the mother and her fetus at risk. Sensitizing staff about this special group of women can create conditions in which women feel respected, heard, and satisfied with their prenatal care.


Pulmonary arteriole remodeling in hypoxic broilers expressing different amounts of endothelial nitric oxide synthase.
Lung samples from 30 pulmonary hypertensive chickens expressing low nitric oxide (NO) synthase activity in endothelial arteriolar cells were compared with samples taken from 30 nonhypertensive animals expressing high activity to investigate a possible relationship between the just-mentioned expressions and pulmonary vascular remodeling. The external diameter and media muscular thickness in 20 arterioles of 50- to 200-microm external diameter were measured in lung samples from 17-, 30-, and 42-d-old broilers to calculate %T (medial wall width divided by external vessel diameter x 100). The %T was higher in hypertensive chickens than in nonhypertensive (P < 0.01) chickens and was inversely related to NO synthase enzyme (P < 0.01). It can be inferred from this study that NO is involved in the remodeling process in broilers with hypoxic pulmonary hypertension syndrome.


Transcription factor nuclear factor-kappa B is activated in neurons after focal cerebral ischemia.
Nuclear factor-kappa B (NF-kappaB) is a multisubunit transcription factor that when activated induces the expression of genes encoding acute-phase proteins, cell adhesion molecules, cell surface receptors, and cytokines. NF-kappaB is composed of a variety of protein subunits of which p50-and p65-kDa (RelA) are the most widely studied. Under resting conditions, these subunits reside in the cytoplasm as an inactive complex bound by inhibitor proteins, IkappaB alpha and IkappaB beta. On activation, IkappaB is phosphorylated by IkappaB kinase and ubiquitinated and degraded by the proteasome; simultaneously, the active heterodimer translocates to the nucleus where it can initiate gene transcription. In the periphery, NF-kappaB is involved in inflammation through stimulation of the production of inflammatory mediators. The role of NF-kappaB in the brain is unclear. In vitro, NF-kappaB activation can be either protective or deleterious. The role of NF-kappaB in ischemic neuronal cell death in vivo was investigated. Adult male rats were subjected to 2 hours of focal ischemia induced by middle cerebral artery occlusion (MCAO). At 2, 6, and 12 hours after reperfusion, the expression and transactivation of NF-kappaB in ischemic versus nonischemic cortex and striatum were determined by immunocytochemistry and by electrophoretic mobility gel-shift analysis. At all time points studied, p50 and p65 immunoreactivity was found exclusively in the nuclei of cortical and striatal neurons in the ischemic hemisphere. The contralateral nonischemic hemisphere showed no evidence of nuclear NF-kappaB immunoreactivity. Double immunofluorescence confirmed expression of p50 in nuclei of neurons. Increased NF-kappaB DNA-binding activity in nuclear extracts prepared from the ischemic hemisphere was further substantiated by electrophoretic mobility gel-shift analysis. Because the activation of NF-kappaB by many stimuli can be blocked by antioxidants in vitro, the effect of the antioxidant, LY341122, previously shown to be neuroprotective, on NF-kappaB activation in the MCAO model was evaluated. No significant activation of NF-kappaB was found by electrophoretic mobility gel-shift analysis in animals treated with LY341122. These results demonstrate that transient focal cerebral ischemia results in activation of NF-kappaB in neurons and supports previous observations that neuroprotective antioxidants may inhibit neuronal death by preventing the activation of NF-kappaB.


Facile one-pot synthesis of Ge/TiO2 nanocomposite structures with improved electrochemical performance.
Germanium (Ge) as an alternative to graphite exhibits a fairly high theoretical energy density and improved Li+ ion diffusivity. However, the seriously deteriorated electrochemical performance of Ge during cycling and the difficulty in the preparation of Ge-based nanostructures can hinder the utilization of Ge as an anode. Thus, in this study, a nanocomposite structure with Ge and TiO2 (Ge/TiO2) was synthesized using a facile one-pot method with different ratios of a Ge source with a dominant GeO2 phase and titanium isopropoxide. From X-ray diffraction, electron microscopy, and X-ray photoelectron spectroscopy, the Ge/TiO2 nanocomposites were found to be spherical structures homogeneously consisting of the reduced Ge as an active material and amorphous TiO2 as a matrix. In particular, the Ge/TiO2 nanocomposite with an appropriate amount of TiO2 exhibited improved electrochemical properties, i.e., a coulombic efficiency of 97% and a retention of 61% for 100 cycles, compared to commercial Ge (a coulombic efficiency of 82% and a retention of 16%). This demonstrates that the amorphous TiO2 matrix could relieve a volumetric expansion of the Ge active material in the nanocomposite electrode generated during the cycling process.


Naturalistic Usability Testing of Inpatient Medication Reconciliation Software.
Medication history errors are common at admission, but can be mitigated through the implementation of medication reconciliation (MR). We designed multi-media software to assist clinicians with collection of an admission history. This manuscript describes a naturalistic usability study conducted on the hospital wards. Our goals were to 1) estimate the impact of our workflow upon departmental productivity and 2) determine the ability of our software to detect discrepancies. We furnished clinical pharmacists with our application on a tablet PC and asked them to collect a bedside history. We used 1) time-motion analysis to estimate cycle-time and 2) chart reviews to estimate error detection rates. Our intervention detected an average of 7.7 discrepancies per admission (11.7 per pharmacy-shift). A panel rated 67% of these discrepancies as 'high' or 'very high' risk. The cycle-time per admission was slightly longer than usual care processes (20.5 min vs. 17.9 min), but included a bedside interview. In general, pharmacists agreed that the technology improved the completeness and accuracy of a medication history. However, workflow leveling strategies are important to implementing a durable process. In conclusion, a pharmacist-mediated, patient-centered technology holds promise for improving the quality of MR and overall clinical performance.


Inhibition airway remodeling and transforming growth factor-β1/Smad signaling pathway by astragalus extract in asthmatic mice.
Airway remodeling is characterized by airway wall thickening, subepithelial fibrosis, increased smooth muscle mass, angiogenesis and increased mucous glands, which can lead to a chronic and obstinate asthma with pulmonary function depression. In the present study, we investigated whether the astragalus extract inhibits airway remodeling in a mouse asthma model and observed the effects of astragalus extract on the transforming growth factor-β1 (TGF-β1)/Smad signaling pathway in ovalbumin-sensitized mice. Mice were sensitized and challenged by ovalbumin to establish a model of asthma. Treatments included the astragalus extract and budesonide. Lung tissues were obtained for hematoxylin and eosin staining and Periodic acid-Schiff staining after the final ovalbumin challenge. Levels of TGF-β1 were assessed by immunohistology and ELISA, levels of TGF-β1 mRNA were measured by RT-PCR, and levels of P-Smad2/3 and T-Smad2/3 were assessed by western blotting. Astragalus extract and budesonide reduced allergen-induced increases in the thickness of bronchial airway and mucous gland hypertrophy, goblet cell hyperplasia and collagen deposition. Levels of lung TGF-β1, TGF-β1 mRNA and P-Smad2/3 were significantly reduced in mice treated with astragalus extract and budesonide. Astragalus extract improved asthma airway remodeling by inhibiting the expression of the TGF-β1/Smad signaling pathway, and may be a potential drug for the treatment of patients with a severe asthma airway.


Timing Is Everything: What Is the Optimal Duration After Chemoradiation for Surgery for Rectal Cancer?
The Oncology Grand Rounds series is designed to place original reports published in the Journal into clinical context. A case presentation is followed by a description of diagnostic and management challenges, a review of the relevant literature, and a summary of the authors' suggested management approaches. The goal of this series is to help readers better understand how to apply the results of key studies, including those published in Journal of Clinical Oncology , to patients seen in their own clinical practice. A 47-year-old woman was referred for management of a newly diagnosed rectal cancer. She presented with a 2-month history of rectal bleeding and change in bowel habits. She underwent a colonoscopy that demonstrated a 5-cm fungating, friable, and partially obstructing mass in the distal rectum, approximately 5 cm from the anal verge. The tumor was palpable on digital rectal examination on the anterior wall of rectum. The biopsy demonstrated a moderately differentiated invasive adenocarcinoma, microsatellite stable. A staging work-up, including a computed tomography scan of the chest, abdomen, and pelvis, demonstrated rectal wall thickening in the midrectum and small lymph nodes in the left perirectal fat. There was a nonspecific 3-mm right lower lobe pulmonary nodule. Rectal magnetic resonance imaging demonstrated a 3-cm mass arising from mid-distal rectum with minimal extension beyond muscularis propria into the mesorectal fat, but without invasion of mesorectal fascia ( Fig 1 ). There were at least three small mesorectal lymph nodes present; the largest rounded node measured up to 5 mm, and no additional pelvic lymphadenopathy was identified. Her carcinoembryonic antigen was 1.1, and all other laboratory studies were within normal limits. She was seen in the Colorectal Multidisciplinary Conference for a discussion of her treatment options.


Effects of stimulus salience on the magnitude of latent inhibition after compound conditioning.
The effect of stimulus salience on latent inhibition (the retardation of conditioning produced by prior exposure to the event to be used as the conditioned stimulus [CS]) was examined in an experiment using rats as subjects and the conditioned suppression procedure. The stimuli were a more salient light and a less salient tone-rats trained with light as the CS showed more suppression than rats trained with the tone as the CS; and rats tested with tone and light separately after conditioning with a CS consisting of a tone + light compound showed more suppression to the light than the tone. This pattern of results was reversed, however, in subjects given a series of nonreinforced presentations of the tone and the light separately prior to conditioning with the compound. We conclude that latent inhibition develops more readily for the more salient stimulus and that its effects can outweigh those that derive from the intrinsic salience of the stimulus. Theories of latent inhibition that predict, or can accommodate, this conclusion are considered.


A new method for studying cell cycle characteristics in ANLL using double-labeling with BrdU and 3HTdr.
Ten patients with acute nonlymphocytic leukemia (ANLL) received bromodeoxyuridine (BrdU) at 100 mg/M2 intravenously over 1 h. BrdU is incorporated into the DNA by S-phase cells and was detected by using a monoclonal anti-BrdU antibody in the bone marrow aspirate (BM) and biopsy specimens obtained at the end of the infusion. Additionally, BM was incubated in vitro with tritiated thymidine (3HTdr) and processed by our previously described double-label method. This allowed us to measure the duration of S-phase (Ts) and total cell cycle time (Tc) of myeloblasts. Data revealed a higher number of S-phase cells from biopsies (21%) than BM (5%). The Ts ranged from 9 to 35 h and Tc ranged between 36 and 152 h in different patients. Using this method, data are available within 48 h and if shown to be clinically relevant, may be useful for prospective planning of therapy in individual patients.


Understanding the Effects of Anesthesia on Cortical Electrophysiological Recordings: A Scoping Review.
General anesthesia in animal experiments is an ethical must and is required for all the procedures that are likely to cause more than slight or momentary pain. As anesthetics are known to deeply affect experimental findings, including electrophysiological recordings of brain activity, understanding their mechanism of action is of paramount importance. It is widely recognized that the depth and type of anesthesia introduce significant bias in electrophysiological measurements by affecting the shape of both spontaneous and evoked signals, e.g., modifying their latency and relative amplitude. Therefore, for a given experimental protocol, it is relevant to identify the appropriate anesthetic, to minimize the impact on neuronal circuits and related signals under investigation. This review focuses on the effect of different anesthetics on cortical electrical recordings, examining their molecular mechanisms of action, their influence on neuronal microcircuits and, consequently, their impact on cortical measurements.


Causal Association Between Periodontitis and Type 2 Diabetes: A Bidirectional Two-Sample Mendelian Randomization Analysis.
Background: Previous observational studies have reported a bidirectional association between periodontitis and type 2 diabetes, but the causality of these relationships remains unestablished. We clarified the bidirectional causal association through two-sample Mendelian randomization (MR). Methods: We obtained summary-level data for periodontitis and type 2 diabetes from several published large-scale genome-wide association studies (GWAS) of individuals of European ancestry. For the casual effect of periodontitis on type 2 diabetes, we used five independent single-nucleotide polymorphisms (SNPs) specific to periodontitis from three GWAS. The summary statistics for the associations of exposure-related SNPs with type 2 diabetes were drawn from the GWAS in the Diabetes Genetics Replication and Meta-analysis (DIAGRAM) consortium and the FinnGen consortium R5 release, respectively. For the reversed causal inference, 132 and 49 SNPs associated with type 2 diabetes from the DIAGRAM consortium and the FinnGen consortium R5 release were included, and the summary-level statistics were obtained from the Gene-Lifestyle Interactions in Dental Endpoints consortium. Multiple approaches of MR were carried out. Results: Periodontitis was not causally related with the risk of type 2 diabetes (all p > 0.05). No causal effect of type 2 diabetes on periodontitis was found (all p > 0.05). Estimates were consistent across multiple MR analyses. Conclusion: This study based on genetic data does not support a bidirectional causal association between periodontitis and type 2 diabetes.


Nightmares: risk factors among the Finnish general adult population.
To identify risk factors for experiencing nightmares among the Finnish general adult population. The study aimed to both test whether previously reported correlates of frequent nightmares could be reproduced in a large population sample and to explore previously unreported associations. Two independent cross-sectional population surveys of the National FINRISK Study. Age- and sex-stratified random samples of the Finnish population in 2007 and 2012. A total of 13,922 participants (6,515 men and 7,407 women) aged 25-74 y. N/A. Nightmare frequency as well as several items related to socioeconomic status, sleep, mental well-being, life satisfaction, alcohol use, medication, and physical well-being were recorded with a questionnaire. In multinomial logistic regression analysis, a depression-related negative attitude toward the self (odds ratio [OR] 1.32 per 1-point increase), insomnia (OR 6.90), and exhaustion and fatigue (OR 6.86) were the strongest risk factors for experiencing frequent nightmares (P < 0.001 for all). Sex, age, a self-reported impaired ability to work, low life satisfaction, the use of antidepressants or hypnotics, and frequent heavy use of alcohol were also strongly associated with frequent nightmares (P < 0.001 for all). Symptoms of depression and insomnia were the strongest predictors of frequent nightmares in this dataset. Additionally, a wide variety of factors related to psychological and physical well-being were associated with nightmare frequency with modest effect sizes. Hence, nightmare frequency appears to have a strong connection with sleep and mood problems, but is also associated with a variety of measures of psychological and physical well-being.


Nitrate Nonutilizing Mutants and Vegetative Compatibility Groups in Fusarium poae
Nitrate nonutilizing (nit) mutants were recovered from 24 isolates of Fusarium poae and used to force heterokaryons between these isolates and to determine vegetative compatibility. Between 30 and 90% of the mycelial blocks, cultured on medium containing chlorate, produced nit mutants. The amount of chlorate in the medium altered the frequency and spectrum of nit mutants recovered. Most of the mutants (63%) had lesions at a nitrate reductase structural locus (nit1). Another 30% were mutants at one or more loci that control the production of a molybdenum-containing cofactor necessary for nitrate reductase activity (NitM). A few (6%) of the mutations occurred in a regulatory gene specific for the nitrate reduction pathway (nit3). Pairings between nit1 and NitM mutants were made on minimal medium containing nitrate as the sole nitrogen source. A mutant grows thinly unless it forms a complementary heterokaryon upon contact with another mutant. Heterokaryon formation was indicated by dense growth where the two mutant colonies touched. The 24 isolates could be divided into 13 nonoverlapping vegetative compatibility groups, suggesting that asexual exchange of genetic information within F. poae is subject to significant limitations.


[Japanese cases of hyperostosis frontalis interna].
Although hyperostosis frontalis interna is common in the western countries, it has been rarely reported in the literature in Japan. We had a chance to observe 5 cases diagnosed as hyperostosis frontalis interna. They were found among 10,902 patients who came to our hospital from August 1, 1993 to September 30, 1995. All the patients in these five cases are females aged 67 to 85 (mean = 74.2 years). Four of the 5 cases had been treated as hypertension, 2 as diabetes mellitus, and 1 as hyperlipoidemia. Two cases were accompanied by unruptured aneurysms. The pathology of one case accompanied by chronic subdural hematoma revealed no apparent development of Haversian systems of bone. It seems that the prevalence of this disease in Japan would increase from now on due to the fact that the life style and the diet among Japanese people has been getting westernized.


Acupuncture for Dyspnea in Lung Cancer: Results of a Feasibility Trial.
Dyspnea is a common and distressing symptom for patients with lung cancer (LC) because of disease burden, therapy toxicity, and comorbid illnesses. Acupuncture is a centuries-old therapy with biological plausibility for relief of dyspnea in this setting. This pilot study aimed to evaluate the feasibility and preliminary effectiveness of acupuncture for dyspnea among patients with LC. Eligible patients had a diagnosis of LC and clinically significant dyspnea without a clear organic cause. The treatment consisted of 10 weekly acupuncture sessions, with a follow-up visit 4 weeks after therapy. The primary outcome was dyspnea severity as measured using a validated Numerical Rating Scale (NRS) of 0 to 10 (10 being "most severe shortness of breath imaginable"). We enrolled 12 patients in the study. The median age was 64.5 years; 66.7% of the patients were female, and 66.7% were Caucasians. Among those enrolled, 10 (83.3%) were able to complete all 10 acupuncture sessions. Acupuncture was well tolerated; adverse events were mild and self-limited. Mean (SD) dyspnea scores on the NRS improved from 6.3 (1.7) at baseline to 3.6 (1.9; P = .003) at the end of treatment and 3.2 (2.3; P = .008) at follow-up. Fatigue and quality of life also improved significantly with acupuncture (P < .05). Among patients with LC, acupuncture was well tolerated and exhibited promising preliminary beneficial effects in the treatment of dyspnea, fatigue, and quality of life. Performing a trial in this population appears feasible.


The procoagulant status. Hypercoagulability as a risk factor of primary and secondary infertility.
Thrombophilia is a disorder that makes patients susceptible to intravascular thrombosis that may increase the risk of developing a pregnancy on a known pathology. The female patient diagnosed with hypoplastic uterus and hereditary thrombophilia had a favorable evolution under properly administered anticoagulant treatment. The homozygous status for the C677T mutation may lead to an increase in plasma homocysteine levels, especially in pregnant women, being an associated risk factor for thrombosis. The risk of developing intravascular thrombosis requires primary prevention measures by adding D-dimers in the early diagnostic algorithm, being the most accurate marker of hypercoagulability and endogenous fibrinolysis. The corroboration of the hypercoagulability status with the results of genotyping, the frequencies of the minor/major alleles studied, single mononucleotide polymorphisms (SNPs) and the establishment of preventive therapy, aims to prevent intravascular thrombosis and thromboembolic phenomena.


Binocular detection of masked patterns in young and old observers.
A visual pattern embedded in noise is detected appreciably better when the stimulus complex contains interocular cues (dichoptic condition) than when such cues are absent (binoptic condition). In a recent study (F. Speranza, G. Moraglia, & B. A. Schneider, 1995) the authors showed that the relative difference between binoptic and dichoptic thresholds does not change with age. However, older adults showed higher binoptic and dichoptic thresholds, thus suggesting an age-related difficulty with degraded stimulation. In this article the authors first replicated these findings and proceeded next to investigating whether age-related changes in processing efficiency, additive internal noise, and the spatial frequency bandwidth of the detecting filters could account, separately or concurrently, for the elevated thresholds in noise exhibited by the older adults. Results indicate that this increase is not attributable to age-related changes in filter bandwidth or internal noise. Rather, the findings can be explained in terms of a decrease in processing efficiency with age.


Measuring impacts of continuing professional development (CPD): The development of the CPD impacts survey (CPDIS).
Evaluating continuing professional development (CPD) programs is essential to demonstrate their value to participants and their institutions, and to inform the improvement and quality of CPD programs. Existing surveys measure a narrow range of impacts. This study developed a survey that can measure a broad range of impacts of CPD programs, not just those that are easily measured such as knowledge, skills, attitudes and confidence. The development of the CPD Impacts Survey (CPDIS) was informed by best practice guidelines. A systematic scoping review, qualitative interviews, and existing survey instruments were used to develop the initial survey items. Past participants from two international health professions education institutes completed the survey (n = 292). Principal component analysis (PCA) was used to refine the survey. The PCA provided a three-component solution. Component 1 (learnings and self-efficacy), Component 2 (networking and building community), and Component 3 (achievement and validation) comprising 47, 14, and 13 items, respectively. The three components had high internal consistency (α = .98, α = .95, α = .92, respectively). The CPDIS is the first survey to assess a broader range of impacts of CPD programs. Given the substantial financial and opportunity costs of participation in CPD, the CPDIS will allow more efficient and accurate evaluation of the utility and value of CPD programs.


Complete androgen insensitivity syndrome: effect on human meibomian gland secretions.
To determine whether androgen receptors affect the fatty acid profiles of neutral and polar lipids in human meibomian gland secretions. Meibomian gland secretion samples were obtained from both eyes of (1) women with complete androgen insensitivity syndrome, a condition characterized by dysfunctional androgen receptors, and (2) age-matched female and male controls. Samples were processed for high-performance liquid chromatography, mass spectrometry, or both and for analysis of the mass spectra of neutral and polar lipid fatty acid fragment ions by 3 different methods. Androgen receptor dysfunction is associated with significant alterations in the appearance of numerous molecular species in the neutral and polar lipid fractions of meibomian gland secretions. The ability to detect these differences, and to assess their nature and extent, was facilitated by the use of several analytic approaches. Sex-related differences exist in the expression of a variety of neutral and, especially, polar fatty acid products in meibomian gland secretions. Androgens exert a significant effect on neutral and polar lipids in human meibomian gland secretions, and these hormonal effects may be mediated through androgen receptors.


On the doublet states of the potassium trimer.
The potassium trimer is investigated in its lowest electronic doublet states, employing several high-level ab initio methods (coupled cluster with single, double, and noniterative triple excitations, multiconfiguration self-consistent field, and multireference Rayleigh-Schrodinger perturbation theory of second order). One-dimensional cuts through the lowest 12 electronic states at C(2v) symmetry give insight in the complex electronic structure of the trimer, showing several (pseudo-)Jahn-Teller distortions that involve two or three excited states. Contour plots of the involved molecular orbitals are shown to prove the validity of the shell model frequently used for a qualitative description of metallic clusters.


Autoantibody to an immunoregulatory inducer population in patients with juvenile rheumatoid arthritis.
The human inducer (T4(+)) and reciprocal cytotoxic/suppressor (T5(+)/T8(+)) subsets have been defined by monoclonal antibodies. In the present study, we examined the relationship of naturally occurring anti-T cell autoantibodies found in patients with active juvenile rheumatoid arthritis (JRA) to these subsets. In one approach, normal T cells were treated with anti-T4 or anti-T8 to eliminate the corresponding subset of cells and then analyzed for reactivity with JRA sera. It was found that JRA sera were reactive with only 15% of an enriched cytotoxic/suppressor population, whereas they reacted with 37% of an enriched inducer population. In reciprocal studies, JRA(+) T cells were eliminated with JRA sera and complement and the residual T cells (JRA(-)) reacted with monoclonal antibodies and indirect immunofluorescence on a fluorescence-activated cell sorter. As expected, the JRA sera and complement treatment of unfractionated T cells markedly diminished the T4(+) subset, whereas there was a concomitant increase in T cells reactive with anti-T5 and anti-T8. A similar diminution in T4(+) T cells was found in the circulating peripheral T cell compartment of patients with active JRA who possessed the JRA antibody. Functional studies demonstrated that removal of the JRA(+) population of T cells diminished phytohemagglutinin and soluble antigen proliferative responses, both of which were previously shown to be functions of T4(+) T cells. More importantly, in the absence of JRA(+) T cells, pokeweed mitogen-stimulated immunoglobulin production was markedly enhanced, despite the concomitant increase in T5(+)/T8(+) cytotoxic/suppressor cells. These results suggest that the JRA serum may define a Qal-like antigen found predominantly on the human inducer population which could activate suppressor and/or other feedback regulatory cells.


Protein acetylation and spermatogenesis.
Spermatogenesis refers to the developmental process of male germ cell formation from the spermatogonial stem cell to mature spermatozoa. The progression of male germ cells through the different phases of development, along with changes in cellular size and morphology, involves a coordinated change in their gene expression program at both the transcript and protein levels. It is well known that the stability, biological activity and cellular localization of proteins are regulated by post-translational modifications. In this review, we provide a brief update of current knowledge about the role of protein acetylation in mammalian spermatogenesis. Based on recent findings specific examples were cited to illustrate how these modifications are involved in controlling the different events that are important to the proper development of male germ cells.


A comparison of long-term outcomes of portal versus systemic venous drainage in pancreatic transplantation: a systematic review and meta-analysis.
Pancreas transplantation venous effluent can be drained via the portal vein or the systemic circulation; however, no recommendation exists for the ideal technique. A systematic review of the literature from 1989 through 2014 using PubMed, CINHAL, and Cochrane Library for portal versus systemic venous drainage was undertaken. Only studies on humans and published in English were considered. Measures of glycemic control and total cholesterol were synthesized for meta-analysis utilizing random-effects models. Of 166 articles retrieved, 15 articles were included for meta-analysis. Patient and graft survival were comparable in a large database study as well as in the only randomized control study. No differences in complications were seen when exocrine drainage was enteric for the systemic venous group. Fasting insulin (-34.13 pmol/mL, p < 0.001) was significantly lower within the portal drained group; however, fasting blood glucose levels (-3.4 mg/dL, p = 0.32) and hemoglobin A1C levels (mean difference 0.124%, p = 0.25) were comparable. Total cholesterol levels (-3.62 mg/dL, p = 0.447), as well as other measures of lipids, showed no difference. Based on this systematic review and meta-analysis, there is no evidence of differences in outcomes or metabolic control in patients undergoing pancreatic transplant with portal venous drainage compared to the systemic venous drainage.


The enrichment of eggs with folic acid through supplementation of the laying hen diet.
In light of evidence supporting a need for humans to increase their dietary folate intakes, experiments were conducted to evaluate the extent to which egg folate levels could be increased. In Study 1, Hyline W36 hens (n = 6/diet) received a barley-based diet, containing 0 or 10 mg/kg of crystalline folic acid, to establish the potential for folate incorporation into table eggs. In Study 2, 70 hens were divided into seven treatment groups (n = 10 hens/diet) and received diets supplemented with 0, 1, 2, 4, 8, 16, or 32 mg folic acid/kg diet. In Study 3, 64 hens received the barley-based diet with or without 4 mg folic acid/kg diet. Eggs were collected and stored for 0, 7, 14, 21, or 28 d, prior to folate determinations. The folate content of eggs was determined by HPLC for 5-methyltetrahydrofolate (the sole form of folate in egg yolk). Results from Study 1 showed that a 10 mg/kg inclusion of folic acid increased folate incorporation into egg yolk (41.0 +/- 0.7 microg /egg) over that of an unsupplemented diet (17.5 +/- 0.7 microg /egg; P = 0.0001). In Study 2, the response of egg folate to dietary folic acid supplementation was saturable, with 90% of maximal egg folate levels established at approximately 4 mg folic acid/ kg diet. Results from Study 3 showed that folate levels are stable, in control and fortified eggs, during 28 d of storage at 4 C. In terms of its nutritional value, one large egg collected from a folic acid-supplemented hen provided approximately 12.5% of the recommended dietary allowance (RDA) for adult humans (RDA = 400 mg/d).


Review of road accident analysis using GIS technique.
The number of road crashes is significantly growing worldwide. In the transportation sector, accident outcomes are usually the loss of lives and injuries. To avoid further damages, a tool entitled geographical information system (GIS) could be helpful. GIS has the most demanding tools used to analyze road accidents and road design that can be noteworthy in traffic accident prevention. The purpose of this review is to propose the superlative approach of GIS applicable to accident analysis in different circumstances. The reviewed statistical results of accidents are performed by GIS but the numerical study is not consummate by GIS. Mainly, four essential GIS techniques are introduced and discussed in this review paper to simulate road accidents and suggest some prolific accident analysis tools for road safety.


Increased concentration of calcium channel and intracellular free calcium in the acute phase of deoxycorticosterone acetate salt hypertension.
Cardiac calcium channels and platelet Ca2+ concentrations were examined in deoxycorticosterone acetate (DOCA)-salt hypertension in rats to clarify the role of calcium channels in the regulation of intracellular Ca2+. Calcium channels were determined by the radioligand binding method and platelet Ca2+ concentrations were measured by a fluorescent Ca2+ indicator, Quin-2. The number of cardiac calcium channels was increased in DOCA-salt hypertension compared with controls 10 days after nephrectomy. Platelet Ca2+ concentrations were also increased in the acute phase of DOCA-salt hypertension. No significant changes were observed in the number of cardiac calcium channels and in platelet Ca2+ concentrations in the chronic phase of DOCA-salt hypertension. These results suggest that the increased platelet Ca2+ concentrations are induced by the increase in the number of calcium channels in the acute phase of hypertension.


[Electrocardiographic abnormalities and cardiovascular risk factors for ischemic heart disease in an adult population from São Paulo, Brazil].
Ischemic heart disease is associated with high rates of mortality and it is the most prevalent among cardiovascular diseases. It is a multifactorial disease, and its prevention depends on control of risk factors. Resting electrocardiogram can be used to detect manifestations of ischemic heart disease, since some electrocardiographic abnormalities are indicative of coronary disease in asymptomatic subjects. The aim of the study was to analyze the relationship between electrocardiographic abnormalities and risk factors for ischemic heart disease in an adult population from the metropolitan area of São Paulo. A cross-sectional study was carried out in an adult population (age 20 years and over) from Cotia, near São Paulo, southeastern of Brazil. The study population comprised 1067 subjects. The subjects were interviewed using a questionnaire. The variables studied were: gender, age, smoking behaviour, physical activity, body mass index, waist-to-hip ratio, blood pressure, diabetes mellitus, blood lipids (total cholesterol, LDL-cholesterol, HDL-cholesterol and triglycerides) and electrocardiographic abnormalities. Electrocardiographic abnormalities were grouped according to the classification: ST-T changes, left ventricular hypertrophy and abnormalities non related to ischemic myocardial disease or without abnormalities. The population was relatively -young (79% younger than 50 years, mean age 39.8 +/- 13.2 years old), and 59.3% were female. The resting electrocardiogram showed ST-T changes in 9.5% of the subjects and left ventricular hypertrophy in 3.3%. Female gender, age over 65 years for women and 55 years for men, diabetes, hypertension, total cholesterol >/= 200 mg/dl, LDL-cholesterol >/=130 mg/dl, Triglycerides >/=200 mg/dl, body mass index >/= 25 Kg/m2 and abnormal waist-to-hip ratio, were positively related to ST-T changes. Male gender, diabetes, hypertension and smoking behaviour were positively related to left ventricular hypertrophy. In the presence of ST-T changes on the ECG, we observed an odds ratio of 3.54 for the diagnosis of systolic hypertension an of 1.83 for total cholesterol >/= 200 mg/dl. In the presence of left ventricular hypertrophy on the ECG, we found an odds ratio of 5.92 for systolic hypertension diagnosis. Among the three most important risk factors correlated to electrocardiographic abnormalities indicative of myocardial ischemia, two can be controlled by nutritional intervention (hypertension and total cholesterol >/= 200 mg/dl). This findings suggest the importance of its early detection. Future studies should determine the predictive value of the electrocardiogram, a simple and inexpensive method, suggesting the presence of cardiovascular risk factors in epidemiological and nutritional studies.


Grunwald-Winstein analysis: isopropyl chloroformate solvolysis revisited.
Specific rates of solvolysis at 25 degrees C for isopropyl chloroformate (1) in 24 solvents of widely varying nucleophilicity and ionizing power, plus literature values for studies in water and formic acid, are reported. Previously published solvolytic rate constants at 40.0 degrees C are supplemented with two additional values in the highly ionizing fluoroalcohols. These rates are now are analyzed using the one and two-term Grunwald-Winstein Equations. In the more ionizing solvents including ten fluoroalcohols negligible sensitivities towards changes in solvent nucleophilicity (l) and very low sensitivities towards changes in solvent ionizing power (m) values are obtained, evocative to those previously observed for 1-adamantyl and 2-adamantyl chloroformates 2 and 3. These observations are rationalized in terms of a dominant solvolysis-decomposition with loss of the CO(2) molecule. In nine of the more nucleophilic pure alchohols and aqueous solutions an association-dissociation mechanism is believed to be operative. Deficiencies in the acid production indicate 2-33% isopropyl chloride formation, with the higher values in less nucleophilic solvents.


Short contact 70% glycolic acid peels as a treatment for photodamaged skin. A pilot study.
Glycolic acid has been one of the more commonly used alpha hydroxy acids for the treatment of photodamaged skin. Its value as a quick "skin refreshing" peeling agent has been widely touted. This type of peel differs from a conventional therapeutic peel (eg, phenol, trichloroacetic acid, or a longer time exposure alpha hydroxy acid peel) in that there is little skin reaction and patients can go about their daily routine without concern. To assess the potential value of glycolic acid-based refresher peels as a cosmetic procedure. Twelve healthy subjects with at least a moderate degree of photodamage were treated with monthly serial 70% glycolic acid peels over a period of 4 months. In addition to the "peels," six subjects were randomized to a 10% glycolic acid-based moisturizer twice daily. Patients were evaluated monthly and graded on a clinical scale using objective measures. No conclusive differences were noted on histologic evaluation. Ninety percent (9/10) of patients felt that overall they noticed significant improvement, however, there was no distinction between the two treatment options. The improvement in fine wrinkling and pigmentation was primarily seen in the patients who additionally received 10% glycolic emollient twice daily. In this limited pilot study, no specific benefit could be assigned to the concomitant use of monthly glycolic acid refresher "peels" in the treatment of photodamaged skin.


Dietary fat, tamoxifen use and circulating sex hormones in postmenopausal breast cancer survivors.
Evidence is inconsistent regarding whether dietary fat influences sex hormone concentrations. This issue is important for breast cancer survivors since clinical recommendations suggest maintaining low hormone levels primarily via pharmacologic agents. This study examines associations between dietary fat and circulating sex hormones among participants in the Health, Eating, Activity and Lifestyle (HEAL) Study, a cohort of breast cancer survivors (N = 511). During a postdiagnosis interview, detailed data were collected on diet, physical activity, lifestyle habits, and medication use (including tamoxifen). Staff measured height and weight and collected fasting bloods. Multivariate linear regression modeled associations of dietary fat with serum sex hormones. Among women using tamoxifen, we observed modest inverse associations of dietary fat with estrone (P < 0.01), estradiol (P < 0.05), testosterone (P < 0.01), free testosterone (P < 0.01), and DHEA (P < 0.01) for higher vs. lower fat intake; but there was no evidence for a trend. Associations were consistent across measures (percent energy from fat, total, saturated, and polyunsaturated fat), and modest effect modification was observed between fat intake and tamoxifen in relation to hormones. Among women not using tamoxifen, fat intake was not associated with hormone concentrations. Further work is needed to confirm the findings and to understand the clinical implications of these observations.


Dentition of eight species of Mediterranean Sea Gobiidae: do dentition characters of gobies reflect phylogenetic relationships?
Oral and pharyngeal dentition was analysed in eight Mediterranean species of five different genera using scanning electron microscopy (SEM). Number, position, shape and size of teeth in the jaws and the pharyngeal tooth plates were used as a basis for comparison among taxa. Three different groups could be established based on the dental morphology among the species investigated and homoplasy due to feeding ecology cannot be considered the reason for similarity among them. The established groups are suggested to reflect phylogenetic relationships and correspond with the scarce published data on the topic.


The complexity of the treatment: the decision-making process among women with rheumatoid arthritis.
There are effective medications available for the treatment of rheumatoid arthritis (RA); yet, medication adherence remains a problem. In this study, grounded theory methodology was used to investigate the decision-making process used by 30 women with RA when deciding to participate in an evidence-based treatment regimen for this disease. From the study findings, a four-phase process was identified. Pain, life functioning, and exhaustion of health care resources are the components of the initial phase, decision initiation. During knowledge acquisition, the second phase, patients attain information about RA and medications used for its treatment from varying sources. The third phase, trusting the health care provider, is defined by a trusting relationship between patients and health care providers. Patients decide to take or not take medications for RA during the final phase, decision is made. The participating women with RA used a complex decision-making process when deciding to take medications for this disease.


Multiple somatotopic representations in the human cerebellum.
The classic view of representation in the cerebellum assumes two homunculi, one in the anterior lobe and one in the posterior lobe. Functional imaging has confirmed this somatotopy in the human anterior lobe but not, so far, in the posterior lobe. Using fMRI, we found separate peaks of activation for finger and toe in three ipsilateral cerebellar regions. In both the anterior and posterior lobe, the toe representation was semicircular around the finger area, with peaks of activation aligned in accord with the classic homunculi. Also, segregated peaks of activation were found in the pyramis vermis. These results confirm the existence of a second homunculus in the posterior lobe of the human cerebellum and suggest a third one.


Dual-layered hydrogels allow complete genome recovery with nucleic acid cytometry.
Targeting specific cells for sequencing is important for applications in cancer, microbiology, and infectious disease. Nucleic acid cytometry (NAC) is a powerful approach for accomplishing this because it allows specific cells to be isolated based on sequence biomarkers that are otherwise impossible to detect. However, existing methods require specialized microfluidic devices, limiting adoption. Here, a modified workflow is described that uses particle-templated emulsification (PTE) and flow cytometry to conduct the essential steps of cell detection and sorting normally accomplished by microfluidics. Our microfluidic-free workflow allows facile isolation and sequencing of cells, viruses, and nucleic acids and thus provides a powerful enrichment approach for targeted sequencing applications.


Serum thyroglobulin concentration in patients with diabetes mellitus.
The serum thyroglobulin (Tg) concentration was measured in 97 patients with diabetes mellitus (39 males, 58 females). Hyper Tg-nemia which exceeds the normal range (1.0-26.6 ng/ml) was observed in 10 patients (3 out of 21 cases treated with diet alone, 3 out of 50 cases treated with oral hypoglycemic agents, 4 out of 26 cases treated with insulin). There was no significant correlation between concentrations of serum Tg and triiodothyronine (T3), thyroxine (T4), fasting plasma glucose (FPG), and hemoglobin A1c (HbA1c). However, a positive correlation was observed between serum concentrations of Tg and thyroid stimulating hormone (TSH). Patients with diabetes were divided into three groups according to the mode of treatment (Group I; diet alone, n = 21, Group II; oral hypoglycemic agents, n = 50, Group III; insulin, n = 26). No significant difference in the serum Tg concentration was observed among the three groups. They were also divided into two groups; normal Tg-nemia (Group A, n = 87) and hyper Tg-nemia (Group B, n = 10). There was no difference between levels of T3, T4, FPG, and HbA1c in the two groups. The serum TSH concentration measured by double antibody RIA and two site immunoradiometric assay in Group B was significantly higher than that in Group A. These results suggest that hyper Tg-nemia in patients with diabetes could be due to the increased TSH concentration which reflects latent subclinical primary hypothyroidism in them.


PsoP1, a milk-clotting aspartic peptidase from the basidiomycete fungus Piptoporus soloniensis.
The first enzyme of the basidiomycete Piptoporus soloniensis, a peptidase (PsoP1), was characterized after isolation from submerged cultures, purification by fractional precipitation, and preparative native-polyarylamide gel electrophoresis (PAGE). The native molecular mass of PsoP1 was 38 kDa with an isoelectric point of 3.9. Similar to chymosin from milk calves, PsoP1 showed a maximum milk-clotting activity (MCA) at 35-40 °C and was most stable at pH 6 and below 40 °C. The complete inhibition by pepstatin A identified this enzyme as an aspartic peptidase. Electrospray ionization-tandem MS showed an amino acid partial sequence that was more homologous to mammalian milk clotting peptidases than to the chymosin substitute from a fungal species, such as the Zygomycete Mucor miehei. According to sodium dodecyl sulfate-PAGE patterns, the peptidase cleaved κ-casein in a way similar to chymosin and hydrolyzed β-casein slowly, as it would be expected from an efficient chymosin substitute.


Primary Failure to an Anti-TNF Agent in Inflammatory Bowel Disease: Switch (to a Second Anti-TNF Agent) or Swap (for Another Mechanism of Action)?
About a third of patients with inflammatory bowel disease do not respond to anti-tumour necrosis factor (anti-TNF) therapy, which is challenging. To review the current data on the two main strategies when facing primary non-response to an anti-TNF agent in inflammatory bowel disease: changing to a second anti-TNF (switching) or to a drug with another mechanisms of action (swapping). We performed a bibliographic search to identify studies reporting on efficacy of any biologic treatment after primary anti-TNF non-response. The efficacy of a second anti-TNF is lower when the reason to withdraw the first one is primary failure. Nevertheless, switching to another anti-TNF even after primary failure may still be effective in some patients. Both vedolizumab and ustekinumab have generally been shown to be less effective in anti-TNF exposed patients. However, despite primary anti-TNF failure, patients may respond to vedolizumab or ustekinumab in a limited but considerable number of cases. The cause for swapping (primary vs. secondary anti-TNF failure) seems to have limited effect on vedolizumab efficacy. Primary anti-TNF non-response seems to be a clearer predictor of treatment failure for ustekinumab. Unfortunately, the two main strategies to treat specifically a patient with primary non-response to an anti-TNF agent-switching to a second anti-TNF or swapping for vedolizumab/ustekinumab-have not been properly compared. The data reviewed in the present study clearly emphasise the imperative need to carry out head-to-head randomised trials in patients exposed to anti-TNF agents in general, and specifically in those with primary non-response to these agents.


Energy Supply for Stomatal Opening in Epidermal Strips of Commelina benghalensis.
The influence of light or darkness on stomatal opening in epidermal strips of Commelina benghalensis was evaluated in the presence or absence of O(2) and/or metabolic inhibitors. Opening was restricted in nitrogen and was promoted by NADH and acids of the tricarboxylic acid cycle (succinate and alpha-ketoglutarate) in CO(2)-free air in light as well as in darkness. The enhancement by light of stomatal opening was prevalent under nitrogen or in the presence of the respiratory inhibitors (sodium azide and oligomycin). Respiratory inhibitors decreased the opening in light or darkness under CO(2)-free air but exhibited no effect under nitrogen, whereas phosphorylation uncouplers were inhibitory in light or darkness under both CO(2)-free air and nitrogen. The results suggest that oxidative phosphorylation is a basic source of energy for stomatal opening, although photophosphorylation could be an energy source.


Improved cryopreservation of chrysanthemum (Chrysanthemum morifolium) using droplet-vitrification.
A droplet-vitrification protocol has been established for cryopreserving Chrysanthemum morifolium cv. Peak using axillary shoot tips and apical shoots of in vitro plants. In the optimized procedure, explants were submitted to a step-wise preculture in liquid sucrose-enriched medium (0.3, 0.5 and 0.7 M for 31,17 and 7 h, respectively). Precultured explants were treated for 40 min with C4 loading solution comprising (w/v) 17.5 percent glycerol + 17.5 percent sucrose, then dehydrated with PVS3 vitrification solution (w/v, 50 percent glycerol + 50 percent sucrose) for 60 min (axillary shoot tips) or 90 min (apical shoots). Explants were cryopreserved by direct immersion in liquid nitrogen in minute drops of PVS3 attached to aluminum foil strips. The optimal age of donor plants was 4-5.5 weeks for apical shoots and 7 weeks for axillary shoot tips, producing post-cryopreservation regeneration percentages of 81.9 percent and 84.9 percernt, respectively. Plants regenerated from cryopreserved samples showed no phenotypical abnormalities and similar profiles of relative DNA content were recorded for control and cryopreserved plants. Our results suggest that the modified droplet-vitrification protocol described in this paper is highly effective and may prove user-friendlier than the cryopreservation protocols already published for chrysanthemum.


Intergenerational risk and resilience pathways from discrimination and acculturative stress to infant mental health.
Preconception and prenatal stress impact fetal and infant development, and women of color are disproportionately exposed to sociocultural stressors like discrimination and acculturative stress. However, few studies examine links between mothers' exposure to these stressors and offspring mental health, or possible mitigating factors. Using linear regression, we tested associations between prenatally assessed maternal acculturative stress and discrimination on infant negative emotionality among 113 Latinx/Hispanic, Asian American, Black, and Multiethnic mothers and their children. Additionally, we tested interactions between stressors and potential pre- and postnatal resilience-promoting factors: community cohesion, social support, communalism, and parenting self-efficacy. Discrimination and acculturative stress were related to more infant negative emotionality at approximately 12 months old (M = 12.6, SD = .75). In contrast, maternal report of parenting self-efficacy when infants were 6 months old was related to lower levels of infant negative emotionality. Further, higher levels of parenting self-efficacy mitigated the relation between acculturative stress and negative emotionality. Preconception and prenatal exposure to sociocultural stress may be a risk factor for poor offspring mental health. Maternal and child health researchers, policymakers, and practitioners should prioritize further understanding these relations, reducing exposure to sociocultural stressors, and promoting resilience.


Preliminary evaluation of besipirdine for the treatment of Alzheimer's disease. Besipirdine Study Group.
Besipirdine hydrochloride (HP 749) is an indole-substituted analog of 4-aminopyridine. Besipirdine enhances both cholinergic and adrenergic neurotransmission in the central nervous system, and may have greater efficacy than purely cholinergic agents in treating dementia due to Alzheimer's disease. The present study examined the efficacy and tolerability of two doses of besipirdine (5 and 20 mg BID) in 275 patients with Alzheimer's disease during 3 months of treatment and during 3 months after withdrawal of treatment. Besipiridine was generally well tolerated. The level of performance on a cognitive test was sustained during 3 months of treatment with besipirdine, whereas the performance of patients treated with placebo deteriorated over the same time period. The results suggest a dose-response relationship, with greater efficacy after 3 months of treatment and longer persistence after treatment withdrawal for besipiridine 20 mg BID than for 5 mg BID. A clinical global rating did not detect a besipirdine treatment effect. The full efficacy after 3 months of treatment did not persist after withdrawal of treatment, suggesting that the benefit is primarily symptomatic. Treatment with higher doses and for longer periods may enhance efficacy on both cognitive and clinical global assessments.


Artificial neural network based approach to EEG signal simulation.
In this paper a new approach to the electroencephalogram (EEG) signal simulation based on the artificial neural networks (ANN) is proposed. The aim was to simulate the spontaneous human EEG background activity based solely on the experimentally acquired EEG data. Therefore, an EEG measurement campaign was conducted on a healthy awake adult in order to obtain an adequate ANN training data set. As demonstration of the performance of the ANN based approach, comparisons were made against autoregressive moving average (ARMA) filtering based method. Comprehensive quantitative and qualitative statistical analysis showed clearly that the EEG process obtained by the proposed method was in satisfactory agreement with the one obtained by measurements.


Continent jejunal reservoir dialysis for end-stage renal disease: is it possible?
With limited organs available for renal transplantation in comparison with the number of patients on the waiting list, and with the drawbacks of dialysis, other forms of treatment for end-stage renal disease (ESRD) need to be investigated. We propose that using a reconfigured segment of bowel as a reservoir in which dialysate of various compositions can be instilled to remove metabolic wastes usually handled by the kidney may augment or replace renal function in a uremic patient. We have chosen the jejunum and have documented our preliminary findings using hyperosmotic dialysate along with the unique characteristics of continent jejunal reservoir dialysis (CJRD). With further refinements, CJRD may eventually be offered as an alternative treatment for ESRD.


Effect of Transplanting Various Concentrations of a Composite of Human Umbilical Cord Blood-Derived Mesenchymal Stem Cells and Hyaluronic Acid Hydrogel on Articular Cartilage Repair in a Rabbit Model.
Mesenchymal stem cells (MSCs) are known to have therapeutic potential for cartilage repair. However, the optimal concentration of MSCs for cartilage repair remains unclear. Therefore, we aimed to explore the feasibility of cartilage repair by human umbilical cord blood-derived MSCs (hUCB-MSCs) and to determine the optimal concentrations of the MSCs in a rabbit model. Osteochondral defects were created in the trochlear groove of femur in 55 rabbits. Four experimental groups (11 rabbits/group) were treated by transplanting the composite of hUCB-MSCs and HA with various MSCs concentrations (0.1, 0.5, 1.0, and 1.5 x 107 cells/ml). One control group was left untreated. At 4, 8, and 16 weeks post-transplantation, the degree of cartilage repair was evaluated grossly and histologically. Overall, transplanting hUCB-MSCs and HA hydrogel resulted in cartilage repair tissue with better quality than the control without transplantation (P = 0.015 in 0.1, P = 0.004 in 0.5, P = 0.004 in 1.0, P = 0.132 in 1.5 x 107 cells/ml). Interestingly, high cell concentration of hUCB-MSCs (1.5×107 cells/ml) was inferior to low cell concentrations (0.1, 0.5, and 1.0 x 107 cells/ml) in cartilage repair (P = 0.394,P = 0.041, P = 0.699, respectively). The 0.5 x 107 cells/ml group showed the highest cartilage repair score at 4, 8 and 16 weeks post transplantation, and followed by 0.1x107 cells/ml group or 1.0 x 107 cell/ml group. The results of this study suggest that transplantation of the composite of hUCB-MSCs and HA is beneficial for cartilage repair. In addition, this study shows that optimal MSC concentration needs to be determined for better cartilage repair.


A method for the assessment of the quality of mitochondrial preparations.
A new experimental approach was applied to the quantitative assessment of the physiological properties of mitochondria. An analytical function (QUAL) was derived from a linear combination of parameters selected by factor correspondence analysis (FCA). The quality of a mitochondrial preparation can be expressed by two numerical values computed from the function QUAL: a mean value and its variance. These statistical parameters can be used for exploring data from planned experiments.


New factors in the antitrust regulation of hospital mergers.
This research considers four recent hospital merger attempts in smaller urban areas where a hospital merger clearly presents antitrust concerns, as determined by the Federal Trade Commission (FTC) or Department of Justice (DOJ). These cases provide an understanding of the more compelling strategies that hospitals are using to offset federal antitrust concerns and the increased number of factors that are being used by the FTC and DOJ to analyze a merger's effect on a market. In particular, the cases illustrate how federal agencies' likelihood of contesting a merger is now based on the merging hospitals' ability to provide strong evidence of substantial savings resulting from a merger. Further, evidence indicates that states are taking on increasing authority for the oversight of hospital mergers. This federalism trend suggests that consumer protection for hospital mergers may be enhanced through a combination of federal and state oversight measures.


Acupuncture: a useful tool for health care in an operational medicine environment.
Acupuncture is a form of medical care that originated in China; it has evolved and progressed over thousands of years to become one of the most commonly used forms of health care throughout the world. Allopathic (Western) medicine has begun to seriously investigate and to use this system only in the past three decades. Although acupuncture's mechanisms for healing are not fully understood, it helps many conditions. Using acupuncture reduces or eliminates the need for expensive medications and the potential risk of adverse events resulting from medications, with cost savings and health benefits to patients. During a deployment of naval combat engineers to Iraq in support of Operation Iraqi Freedom, acupuncture was used in the health care of sailors, Marines, and soldiers. It objectively and subjectively improved the health of troops in the field. Troops were able to function while being treated, reducing or avoiding sick in quarters or light limited duty status and saving operational man-days. Acupuncture in the right hands can serve as a health force multiplier (amplifying a provider's clinical impact) and can be integrated into routine health care, whether in garrison or in the field.


[Evaluation of loretadine therapy by rhino-manometry in patients with allergic rhinitis].
The authors evaluated the nasal flow and resistance by rhinomanometry in 21 children suffering from ragweed allergy during the season. After this process the loratadine was administered for 1 month. During the therapy the nasal parameters were checked at 2 and 4 weeks, again. The loratadine significantly decreased the nasal resistance in 18 children and improved the nasal airflow in 17 cases. In the second part of the examination 15 ragweed sensitized children were undergone nasal allergen-specific challenge after 40 minutes taken loratadine. The nasal parameters were checked after 30 minutes and 6-8 hours. The statistical analyzes proved that loratadine can prevent and inhibit the early and late-phase of Type I hypersensitivity in contrast of the 12 children who were challenged without loratadine administration. They showed nasal obstruction. Based on the results, loratadine can be considered very effective antihistamine-antiallergenic drug which improves the quality of life of the allergic patients.


[Pathogenicity of Ureaplasma urealyticum serotype 3 in genital tract of mice].
To study the pathogenicity of Ureaplasma urealyticum serotype 3 (UU3) with different concentration in the genital tract of the mice. A total of 156 Kunming mice were divided into 4 groups randomly, including group A, B, C (48 mice in every experimental group) and control group (12 mice in control group). UU 3 at concentration of 1x10(7) copy/g (group A), 1x10(6) copy/g (group B), 1x10(5) copy/g (group C) were inoculated into 48 mice in every experimental group intravaginally, in the mean time, culture medium of UU was given into 12 mice in control group. They were necropsied at 1, 3, 7, 14, 21, 35 days of postinoculation randomly, which included 8 mice of every experimental group and 2 mice of control group every time, and to detect UU3 expression from cervical secretions by FQ-PCR and observing the pathogenicity rate in tissues of cervix, endometrium, fallopian tube by light microscope and calculate the morbidity rate. (1) The total positive rates of UU3 were 63% (30/48) in group A, 50% (24/48) in group B, 17% (8/48) in group C, which showed a significant difference (P<0.01). And at 1, 3, 7, 14, 21, 35 days of postinoculation, the positive rates of group A were 8/8, 7/8, 6/8, 5/8, 4/8 and 0, group B were 7/8, 5/8, 5/8, 4/8, 3/8 and 0, group C were 3/8, 2/8, 2/8, 1/8, 0 and 0; all mice in control group were zero. At all time points, there were statistical difference in the positive rate among three experimental groups only at 1 day (P<0.05). (2) In the positive mice, their UU3 quantity concentration at 1, 3, 7, 14, 21 days were 1.70x10(7), 8.26x10(6), 4.04x10(6), 2.86x10(6), and 2.41x10(5) copy/g in group A; 3.75x10(6), 2.56x10(6), 1.37x10(6), 6.72x10(5), and 1.12x10(5) copy/g in group B, and 1.45x10(5), 1.07x10(5), 5.43x10(4), 4.68x10(3), and 0 copy/g in group C. There were statistical difference among experimental groups at all time points except 21 days (P<0.05). Comparing the concentration among all time points of every group, both group A and B showed a significant difference (P<0.05), group C didn't reach it (P>0.05). (3) The total pathogenicity rates of three groups were significant different at 7-35 days, which were 56% (18/32) in group A, 44% (14/32) in group B, 6% (2/32) in group C (P<0.01). And at 7, 14, 21, 35 days of postinoculation, the pathogenicity rates in group A were 5/8, 5/8, 4/8 and 4/8, group B were 4/8, 4/8, 3/8 and 3/8, group C were 1/8, 0, 1/8 and 0; all mice in control group were zero, which demonstrated significant difference only at 14 days (P<0.05), no other statistical difference were observed (P>0.05). The pathogenicity of UU3 varies with different concentration in genital tract of mice. When UU3 concentration is more than 1x10(6) copy/g, the susceptibility to infection was intensified significantly.


Dissociation vs Repression: A New Neuropsychoanalytic Model for Psychopathology.
Recent research on trauma, attachment and neuroscience point at a clear divide in psychopathology between disorders based on repression, (as in Freud's repression model) and psychopathologies structured on dissociative mechanisms, a response to severe interpersonal trauma. Pathologies based on repression are typical of a neurotic structure, (with better developmental outcome), while pathologies based on dissociation are of more severe, often borderline nature, as in Otto Kernberg's borderline organization (1975). Neurobiology of attachment and affect regulation theory (Allan Schore), developmental psychopathology (Giovanni Liotti) and contemporary relational psychoanalysis (Philip Bromberg), all provide clinical evidence that the most severe psychopathology is of dissociative structure. This paper clarifies the after-effects of first level of traumatization of human agency (i.e., lack of attunement) and of the second level as in cases with actual abuse, maltreatment or incest (Mucci, 2013), with the internalization of a dyad victim/persecutor within the self of the survivor, as seen in borderline psychopathology (Mucci, 2018).


[Menopause in 2004: "hormone replacement therapy" is not what it used to be anymore].
The data concerning post-menopausal hormone replacement therapy (HRT) were recently completely modified. The aim of this review is to present the last studies about post-menopausal HRT and to describe new alternatives to this treatment. In May 2002, the women's health initiative (WHI) trial of post-menopausal HRT was interrupted earlier than expected. The studied hormonal formulation in this arm of the WHI trial was the association of conjugated equine estrogens and medroxyprogesterone. The reason for termination was an increased risk of breast cancer and myocardial infarction in the hormone-therapy group. Later, reports confirmed that this type of HRT could not be used any more for the primary prevention of coronary heart disease even if the absolute risk remained low. There is an increased risk for venous thromboembolism with post-menopausal estroprogestative replacement. This risk does not seem to exist with transdermal estrogens. The other WHI findings concerned the lack of protection against dementia and cognitive decline. On the contrary, osteoporotic hip fractures and colorectal cancers were reduced in the treated group. In April 2004, the estrogen only arm of the same WHI study was also prematurely interrupted because of an increase in the incidence of stroke. The risk of breast cancer was on the contrary not increased after 6.8 years, raising the question of the eventual role of progestins. The impact of the WHI trial on clinical practice was very important since then. The "Agence Francaise de sécurité sanitaire des produits de santé" (AFSSAPS) edited in May 2004 a public recommendation limiting indication for HRT to patients with severe climacteric symptoms. The treatment must now be prescribed for the shortest time and at the minimal dose. The patient has to be precisely informed about the risks with HRT and the practitioner has to re-evaluate his prescription annually. Hormonal or non-hormonal alternatives have also to be considered as phytoestrogens and tibolone for hot flashes, and raloxifene and diphosphonates for osteoporosis prevention. In any case, a healthy diet, exercise and smoking cessation should be encouraged.


Efficient capture of ReO4- on magnetic amine-functionalized MIL-101(Cr): Revealing from selectivity to mechanism.
The efficient decontamination of pertechnetate (99TcO4-) is an essential task for managing radioactive 99Tc in nuclear wastes. Perrhenate, (ReO4-), as a nonradioactive analog, exhibits almost identical physicochemical properties as 99TcO4-. Herein, a novel magnetic amine-functionalized MIL-101(Cr) (NH2-MIL-101(Cr)@Fe3O4) was prepared and used to efficiently remove ReO4- from solution for the facile magnetic separation. A series of environmental parameters were considered to investigate the adsorption performance of NH2-MIL-101(Cr)@Fe3O4. Experimental results suggested that NH2-MIL-101(Cr)@Fe3O4 has reached a satisfied adsorption capacity (~401 mg/g) and a very fast adsorption kinetics at pH 7.0. The selectivity for ReO4- was maintained even in the presence of interfering anions with relatively high concentrations. ReO4- were mainly captured by N-donor sites of the surface-decorated amine via complexation and were trapped in the cavities of modified MIL-101(Cr). NH2-MIL-101(Cr)@Fe3O4 exhibits satisfactory adsorption performance for ReO4- and can be conveniently separated from wastewaters after adsorption.


[Prevention of colorectal cancer].
Colorectal cancer (CRC) is a public health problem. It is one of the most common cancers and mortality rate is around 50%. This article reports on the various methods of primary prevention testing for the population at average risk of developing CRC. Given its slow evolution through pre-cancerous lesions, it is appropiate to identify patients at medium risk and monitore those at high risk. Current screening methods show very different efficiencies. The most efficient are invasive and limit public support. New non-invasive tests based on fecal and blood biomarkers are being developed and will probably help to improve CRC screening in the future in an attempt to lower mortality rate.


Genome-scale CRISPR screens identify host factors that promote human coronavirus infection.
The COVID-19 pandemic has resulted in 275 million infections and 5.4 million deaths as of December 2021. While effective vaccines are being administered globally, there is still a great need for antiviral therapies as antigenically novel SARS-CoV-2 variants continue to emerge across the globe. Viruses require host factors at every step in their life cycle, representing a rich pool of candidate targets for antiviral drug design. To identify host factors that promote SARS-CoV-2 infection with potential for broad-spectrum activity across the coronavirus family, we performed genome-scale CRISPR knockout screens in two cell lines (Vero E6 and HEK293T ectopically expressing ACE2) with SARS-CoV-2 and the common cold-causing human coronavirus OC43. Gene knockdown, CRISPR knockout, and small molecule testing in Vero, HEK293, and human small airway epithelial cells were used to verify our findings. While we identified multiple genes and functional pathways that have been previously reported to promote human coronavirus replication, we also identified a substantial number of novel genes and pathways. The website https://sarscrisprscreens.epi.ufl.edu/ was created to allow visualization and comparison of SARS-CoV2 CRISPR screens in a uniformly analyzed way. Of note, host factors involved in cell cycle regulation were enriched in our screens as were several key components of the programmed mRNA decay pathway. The role of EDC4 and XRN1 in coronavirus replication in human small airway epithelial cells was verified. Finally, we identified novel candidate antiviral compounds targeting a number of factors revealed by our screens. Overall, our studies substantiate and expand the growing body of literature focused on understanding key human coronavirus-host cell interactions and exploit that knowledge for rational antiviral drug development.


Measurable residual disease in hematopoietic stem cell transplantation-eligible patients with acute myeloid leukemia: clinical significance and promising therapeutic strategies.
Allogeneic hematopoietic stem cell transplantation (alloHCT) results in improved outcomes for acute myeloid leukemia (AML) patients compared to consolidation chemotherapy in transplant-eligible patients with adverse-risk disease. Despite achieving a complete remission (CR) to initial therapy, defined as <5% bone marrow blasts with recovery of peripheral blood elements, a large number of patients suffer disease relapse following alloHCT. There is a growing focus on minimal or measurable residual disease (MRD) in patients with AML, variably defined using multiparametric flow cytometry and nucleic acid sequencing techniques, to detect levels of disease not included in current CR terminology. MRD has emerged as an important prognostic tool in AML and may improve risk stratification, further refine selection of post-remission treatment, and provide an opportunity for therapeutic intervention. Herein, we review MRD detection methods, timing of measurement, prognostic implications, and MRD-directed therapy in AML patients undergoing alloHCT.


Dual reward prediction components yield Pavlovian sign- and goal-tracking.
Reinforcement learning (RL) has become a dominant paradigm for understanding animal behaviors and neural correlates of decision-making, in part because of its ability to explain Pavlovian conditioned behaviors and the role of midbrain dopamine activity as reward prediction error (RPE). However, recent experimental findings indicate that dopamine activity, contrary to the RL hypothesis, may not signal RPE and differs based on the type of Pavlovian response (e.g. sign- and goal-tracking responses). In this study, we address this discrepancy by introducing a new neural correlate for learning reward predictions; the correlate is called "cue-evoked reward". It refers to a recall of reward evoked by the cue that is learned through simple cue-reward associations. We introduce a temporal difference learning model, in which neural correlates of the cue itself and cue-evoked reward underlie learning of reward predictions. The animal's reward prediction supported by these two correlates is divided into sign and goal components respectively. We relate the sign and goal components to approach responses towards the cue (i.e. sign-tracking) and the food-tray (i.e. goal-tracking) respectively. We found a number of correspondences between simulated models and the experimental findings (i.e. behavior and neural responses). First, the development of modeled responses is consistent with those observed in the experimental task. Second, the model's RPEs were similar to dopamine activity in respective response groups. Finally, goal-tracking, but not sign-tracking, responses rapidly emerged when RPE was restored in the simulated models, similar to experiments with recovery from dopamine-antagonist. These results suggest two complementary neural correlates, corresponding to the cue and its evoked reward, form the basis for learning reward predictions in the sign- and goal-tracking rats.


Transcriptome Analysis Identifies Novel Prognostic Genes in Osteosarcoma.
Osteosarcoma (OS), a malignant primary bone tumor often seen in young adults, is highly aggressive. The improvements in high-throughput technologies have accelerated the identification of various prognostic biomarkers for cancer survival prediction. However, only few studies focus on the prediction of prognosis in OS patients using gene expression data due to small sample size and the lack of public datasets. In the present study, the RNA-seq data of 82 OS samples, along with their clinical information, were collected from the TARGET database. To identify the prognostic genes for the OS survival prediction, we selected the top 50 genes of contribution as the initial candidate genes of the prognostic risk model, which were ranked by random forest model, and found that the prognostic model with five predictors including CD180, MYC, PROSER2, DNAI1, and FATE1 was the optimal multivariable Cox regression model. Moreover, based on a multivariable Cox regression model, we also developed a scoring method and stratified the OS patients into groups of different risks. The stratification for OS patients in the validation set further demonstrated that our model has a robust performance. In addition, we also investigated the biological function of differentially expressed genes between two risk groups and found that those genes were mainly involved with biological pathways and processes regarding immunity. In summary, the identification of novel prognostic biomarkers in OS would greatly assist the prediction of OS survival and development of molecularly targeted therapies, which in turn benefit patients' survival.


Japan's Slow Response to Improve Access to Inpatient Care for COVID-19 Patients.
The coronavirus disease 2019 (COVID-19) pandemic has exposed various weaknesses in national healthcare systems across the globe. In Japan, this includes the inability to promptly mobilize the resources needed to provide inpatient care in response to the rapidly increasing number of patients. Combined with unclear entry points to healthcare, particularly in emergency cases, this has led to a situation in which access to healthcare is rapidly deteriorating. This study examined problems in Japan's healthcare delivery system. While Japan's healthcare resources (e.g., hospital beds and medical personnel) are comparable to those found in other high-income countries, progress has been slow in securing beds for COVID-19 patients. In addition, the number of beds has only recently reached the levels seen in Western countries. Factors related to slow resource allocation include dispersed existing medical resources (mainly in the private sector), the lack of collaboration mechanisms among private-dominant healthcare providers and public health agencies, an inadequate legal framework for resource mobilization, the insufficient quantification of existing resources, and undesignated entry points to healthcare systems. To better prepare for future disasters, including the next wave of COVID-19, Japan urgently needs to restructure its legal framework to promptly mobilize resources, accurately quantify existing resources, introduce coordination mechanisms with functional differentiations among all community stakeholders, and clearly designate entry points to healthcare.


Acute cytotoxic effects of mercuric compounds in cultured astrocytes prepared from cerebral hemisphere and cerebellum of newborn rats.
We investigated acute cytotoxic effects and Hg accumulation after exposure to methylmercury (MeHg) or Hg(2+) in the presence or absence of serum in cultured astrocytes prepared from the cerebral hemisphere or cerebellum of newborn rats. Dose-related changes in viable cell numbers after exposure to mercuric compounds were not different between astrocytes from both regions under the specified conditions. Accumulation of each compound for 3 h was similar in both astrocytes but that for 24 h became different, especially that of Hg(2+). In both astrocytes, susceptibility to the respective compounds was higher in the order of those exposed immediately after, without, and 24 h after changing the serum-containing medium to a serum-free defined medium (SFDM). Accumulation for 3 h was higher in the respective astrocytes exposed to MeHg or Hg(2+) immediately after being maintained in SFDM than in those exposed 24 h after. These results suggest that accumulation of mercuric compounds up to 3 h strongly correlates with susceptibility, at least when maintained in SFDM. Astrocytic morphology changed to a satellite shape after the medium change to SFDM particularly in cerebellar astrocytes but only a few in cerebral hemisphere astrocytes, and it was reverted to a polygonal shape by MeHg but not Hg(2+) at 3 microM. The present results suggest that although some properties such as morphological changes and Hg accumulation are different between cerebral hemisphere and cerebellar astrocytes, these differences are not simply reflected by susceptibility to the acute cytotoxicity of mercuric compounds.


Control of activating transcription factor 4 (ATF4) persistence by multisite phosphorylation impacts cell cycle progression and neurogenesis.
Organogenesis is a highly integrated process with a fundamental requirement for precise cell cycle control. Mechanistically, the cell cycle is composed of transitions and thresholds that are controlled by coordinated post-translational modifications. In this study, we describe a novel mechanism controlling the persistence of the transcription factor ATF4 by multisite phosphorylation. Proline-directed phosphorylation acted additively to regulate multiple aspects of ATF4 degradation. Stabilized ATF4 mutants exhibit decreased β-TrCP degron phosphorylation, β-TrCP interaction, and ubiquitination, as well as elicit early G(1) arrest. Expression of stabilized ATF4 also had significant consequences in the developing neocortex. Mutant ATF4 expressing cells exhibited positioning and differentiation defects that were attributed to early G(1) arrest, suggesting that neurogenesis is sensitive to ATF4 dosage. We propose that precise regulation of the ATF4 dosage impacts cell cycle control and impinges on neurogenesis.


New frontiers in suicide vulnerability: immune system and sex hormones.
Suicide is one of the leading causes of death worldwide and men have a higher risk of attempting and completing suicide than women. Accumulating evidence leads to a possible key role of the immune system and sex hormones in psychiatric conditions associated with suicide vulnerability (e.g., major depressive disorder). Moreover, the literature highlights a dysregulation of the immune system and altered sex hormone levels in suicidal patients. Sex hormones and the immune system may have a role in suicide risk and sex differences in suicide vulnerability. This brief review emphasises a research area focused on a possible interplay between the immune system and sex hormones that may help develop a better understanding of suicide vulnerability in the perspective of sex-specific therapeutic approaches.


Are we Athens or Florence? COVID-19 in historical context.
We have been here before. In 430 BCE, a plague struck Athens, killing as much as 25% of the population. In 1347 CE, the bubonic plague afflicted western Europe for 4 years, killing as much as 50% of the population. The plague of Athens led to a collapse of their religion, cultural norms and democracy. In contrast, the bubonic plague led eventually to the Renaissance, a growth of art, science and humanism. As we contend with the COVID-19 global pandemic, will we become Athens or Florence?


Clinical and Public Health Implications of HIV- Genetic Diversity and Drug Resistance Mutations in Angola: A Systematic Review.
HIV-1 genetic diversity and drug resistance mutations (DRMs) remain a public health concern mainly in low- and middle-income countries. In this review, we estimated the HIV-1 molecular evolution over the past 40 years (1980-2019) in Angola to help guide affordable strategies for HIV-1 epidemic surveillance. We searched for studies written in English or Portuguese on HIV-1 diversity and DRMs carried out in Angola and published between 1980 and 2019. This review yielded eight studies describing a total of 493 samples. No HIV-1 Group N, O, and P were identified, whereas a ll non-B subtypes f rom Group M were identified. About 66% of HIV-1 subtypes were pure subtype and 34% recombinant strains. The frequency of recombinant strains increases from 1980 to 2019 (23.6%-41.4%, p<0.001). The subtypes C, F1, CRF02_AG, and the recombinant U/H were the most frequent. One DRM in the PIs was found (I54 M), 22 in the nucleoside reverse transcriptase inhibitors (NRTIs), and 18 in the non-nucleoside reverse transcriptase inhibitors (NNRTIs). The major DRM in the NRTIs was the M184V, whereas the G190A, K103N, and Y181C were the major DRMs in the NNRTIs. Over the past 40 years, the frequency of the DRM M184V (50-64.3%, p=0.363), G190A (17.2-46.2%, p=0.021), and K103N (34.5-42.3%, p=0.551) increased, while the frequency of Y181C (17.2-7.7%, p=0.289) decreased. The current review shows an increase in HIV-1 genetic complexity and DRMs in Angola. Our findings suggest the need to include PIs or integrase strand transfer inhibitors in the first-line antiretroviral therapy regimens in Angola.


Training Community-Based Clinicians in Screening and Brief Intervention for Substance Abuse Problems: Translating Evidence into Practice.
Screening and brief intervention in general health care settings are efficacious but have not been widely adopted. Our objective was to assess the effect of an educational intervention on clinicians' substance abuse-related clinical practices. The study was a telephone survey of practicing physicians, nurses, psychologists, physician's assistants, and social workers who attended a half-day continuing education course on one of four occasions. The course covered the stages of behavioral change and motivational counseling, using primarily role play with standardized patients. Of 87 course attendees, 70 (80%) completed the interview. Months to years after the course, most (91%) reported that the course made an impact on their practice. Most (78%) of respondents reported that they frequently or always asked new patients who drank alcohol a formal screening questionnaire such as the CAGE, and 94% frequently or always assessed their substance abusing patients' readiness to change. Most respondents reported that since taking the course they were more likely (1) to screen patients for alcohol or drug related problems (86%) and (2) to ask patients about their substance abuse on a follow-up visit (96%). After exposure to an active-learning half-day continuing education course, clinicians reported improvement with and high rates of desirable substance abuse-related clinical practices up to 5 years later. Continuing education efforts that incorporate active learning directed toward practicing clinicians show promise for improving rates of brief intervention for alcohol and other drug abuse.


Solid lipid nanoparticles for applications in gene therapy: a review of the state of the art.
Gene therapy represents a new paradigm in the prevention and treatment of many inherited and acquired diseases, including genetic disorders, such as cystic fibrosis, haemophilia and many somatic diseases, such as tumours, neurodegenerative diseases and viral infections, such as AIDS. Among a large array of non-viral transfection agents used for in-vitro applications, cationic SLNs are the topic of this review, being recently proposed as an alternative carrier for DNA delivery, due to many technological advantages such as large-scale production from substances generally recognized as safe, good storage stability and possibility of steam sterilization and lyophilisation. The authors give some information on the knowledge of intracellular trafficking and SLNs-DNA complex chemical-physical properties reported until now in the literature. The future success of cationic SLNs for administration of genetic material will depend on their ability to efficiently cross the physiological barriers, selectively targeting a specific cell type in vivo and expressing therapeutic genes.


Subproteomics: identification of plasma membrane proteins from the yeast Saccharomyces cerevisiae.
As a consequence of their poor solubility during isoelectric focusing, integral membrane proteins are generally absent from two-dimensional gel proteome maps. In order to analyze the yeast plasma membrane proteome, a plasma membrane purification protocol was optimized in order to reduce contaminating membranes and cytosolic proteins. Specifically, the new fractionation scheme largely depleted the plasma membrane fraction of cytosolic proteins by deoxycholate stripping and ribosomal proteins by sucrose gradient flotation. The plasma membrane complement was resolved by two-dimensional electrophoresis using the cationic detergent cetyl trimethyl ammonium bromide in the first, and sodium dodecyl sulfate in the second dimension, and fifty spots were identified by matrix-assisted laser desorption/ionization-time of flight mass spectometry. In spite of the presence of still contaminating ribosomal proteins, major proteins corresponded to known plasma membrane residents, the ABC transporters Pdr5p and Snq2p, the P-type H(+)-ATPase Pma1p, the glucose transporter Hxt7p, the seven transmembrane-span Mrh1p, the low affinity Fe(++) transporter Fet4p, the twelve-span Ptr2p, and the plasma membrane anchored casein kinase Yck2p. The four transmembrane-span proteins Sur7p and Nce102p were also present in the isolated plasma membranes, as well as the unknown protein Ygr266wp that probably contains a single transmembrane span. Thus, combining subcellular fractionation with adapted two-dimensional electrophoresis resulted in the identification of intrinsic plasma membrane proteins.