Assessment of therapeutic drug monitoring of vancomycin in elderly patients according to new guidelines.
Concerns regarding increasing microbial resistance to vancomycin have resulted in recommendations for a higher trough serum vancomycin concentration. This study aimed to assess the dosage guidelines targeting vancomycin trough concentrations of 15-20 mg/L. About 216 adult patients (age, >60 yr) were treated with intravenous vancomycin. The patients were divided into 2 groups according to their target vancomycin trough concentrations: the previous guideline group (n=108) treated with targeted vancomycin trough concentrations of 5-15 mg/L from Jan 2009 through April 2011 and the new guideline group (n=108) treated with targeted concentrations of 15-20 mg/L from November 2011 through July 2012. The 2 groups were not significantly different with respect to age, weight, initial serum creatinine, initial creatinine clearance, predictive trough levels, doses, serum drug concentrations, and area under the curve/minimal inhibitory concentrations. Regarding the proportions of vancomycin trough concentrations, the target range was achieved in 50% in the previous guideline group and in 16% in the new guideline group. In the previous and new guideline groups, the trough concentrations of 10-20 mg/dL were observed in 32.4% and 52.8% patients, respectively, and those of <10 mg/L were observed in 45.4% and 29.6%, respectively. Compared to the previous guideline group, the new guideline group showed higher proportions in the therapeutic range of 10-20 mg/L and lower proportions in trough concentrations <10 mg/L. The strictly managed vancomycin therapeutic drug monitoring in the new guideline group was assessed as more effective.


Na+-activated K+ current contributes to postexcitatory hyperpolarization in neocortical intrinsically bursting neurons.
The ionic mechanisms underlying the termination of action-potential (AP) bursts and postburst afterhyperpolarization (AHP) in intrinsically bursting (IB) neocortical neurons were investigated by performing intracellular recordings in thin slices of rat sensorimotor cortex. The blockade of Ca(2+)-activated K(+) currents enhanced postburst depolarizing afterpotentials, but had inconsistent and minor effects on the amplitude and duration of AHPs. On the contrary, experimental conditions resulting in reduction of voltage-dependent Na(+) entry into the cells caused a significant decrease of AHP amplitude. Slice perfusion with a modified artificial cerebrospinal fluid in which LiCl (40 mM) partially replaced NaCl had negligible effects on the properties of individual APs, whereas it consistently increased burst length and led to an approximately 30% reduction in the amplitude of AHPs following individual bursts or short trains of stimulus-induced APs. Experiments performed by partially replacing Na(+) ions with choline revealed a comparable reduction in AHP amplitude associated with an inhibition of bursting activity. Moreover, in voltage-clamp experiments carried out in both in situ and acutely isolated neurons, partial substitution of extracellular NaCl with LiCl significantly and reversibly reduced the amplitude of K(+) currents evoked by depolarizing stimuli above-threshold for Na(+)-current activation. The above effect of Na(+)-to-Li(+) substitution was not seen when voltage-gated Na(+) currents were blocked with TTX, indicating the presence of a specific K(+)-current component activated by voltage-dependent Na(+) (but not Li(+)) influx. The above findings suggest that a Na(+)-activated K(+) current recruited by the Na(+) entry secondary to burst discharge significantly contributes to AHP generation and the maintenance of rhythmic burst recurrence during sustained depolarizations in neocortical IB neurons.


Silicon-mediated organic synthesis.
Organic chemists have used the chemical properties of tetracovalent silicon in a remarkable variety of new synthetic transformations. In carbon-functional silanes, exceptional stabilization is provided to a carbocation center in the beta position when the carbon-silicon bond lies in plane. This phenomenon directs electrophilic attack to the silicon-substituted carbon in aryl-, vinyl-, and alkynylsilanes and to carbon-3 in allylsilanes. For different reasons, silicon also stabilizes a carbon-metal bond in the alpha position. Consequently, access to many silicon-containing organometallics is readily available. The exceptional strength of silicon-oxygen and silicon-fluorine bonds is yet another factor that controls the chemical reactivity of silicon reagents. In recent developments, preparative chemists have taken advantage of these properties in imaginative and useful ways.


A low-cost microplasma generation unit allowing for the on-site processing of ZnO-based gas sensors.
In this study, a low-cost gas-sensing device that integrates a zinc-oxide (ZnO)-based gas sensor with a microplasma generation unit is presented. By the proper connection of different sets of electrodes to the desired power sources or the measurement circuit, the device can be operated in the microplasma mode (MPM) or the gas-sensing mode (GSM), respectively. Due to the device design, microplasma can be generated at the same location where the precursor for the sensing materials, namely Zn(NO3)2 solution, is coated. Such an arrangement allows for low temperature and the on-site conversion of the precursor materials to ZnO particles upon microplasma treatment under MPM. GSM can be used to detect ethanol vapor with a concentration between 100 and 18 000 ppm at room temperature. We observed that the sensor can be operated under a wide range of ethanol concentrations with stable sensing behavior for over 80 test cycles.


High resolution SNP array profiling identifies variability in retinoblastoma genome stability.
Both hereditary and nonhereditary retinoblastoma (Rb) are commonly initiated by loss of both copies of the retinoblastoma tumor suppressor gene (RB1), while additional genomic changes are required for tumor initiation and progression. Our aim was to determine whether there is genomic heterogeneity between different clinical Rb subtypes. Therefore, 21 Rb tumors from 11 hereditary patients and 10 nonhereditary Rb patients were analyzed using high-resolution single nucleotide polymorphism (SNP) arrays and gene losses and gains were validated with Multiplex Ligation-dependent Probe Amplification. In these tumors only a few focal aberrations were detected. The most frequent was a focal gain on chromosome 2p24.3, the minimal region of gain encompassing the oncogene MYCN. The genes BAZ1A, OTX2, FUT8, and AKT1 were detected in four focal regions on chromosome 14 in one nonhereditary Rb. There was a large difference in number of copy number aberrations between tumors. A subset of nonhereditary Rbs turned out to be the most genomic unstable, while especially very young patients with hereditary Rb display stable genomes. Established Rb copy number aberrations, including gain of chromosome arm 1q and loss of chromosome arm 16q, turned out to be preferentially associated with the nonhereditary Rbs with later age of diagnosis. In contrast, copy number neutral loss of heterozygosity was detected mainly on chromosome 13, where RB1 resides, irrespective of hereditary status or age. Focal amplifications and deletions and copy number neutral loss of heterozygosity besides chromosome 13 appear to be rare events in retinoblastoma.


MAK33 antibody light chain amyloid fibrils are similar to oligomeric precursors.
Little structural information is available so far on amyloid fibrils consisting of immunoglobulin light chains. It is not understood which features of the primary sequence of the protein result in fibril formation. We report here MAS solid-state NMR studies to identify the structured core of κ-type variable domain light chain fibrils. The core contains residues of the CDR2 and the β-strands D, E, F and G of the native immunoglobulin fold. The assigned core region of the fibril is distinct in comparison to the core identified in a previous solid-state NMR study on AL-09 by Piehl at. al, suggesting that VL fibrils can adopt different topologies. In addition, we investigated a soluble oligomeric intermediate state, previously termed the alternatively folded state (AFS), using NMR and FTIR spectroscopy. The NMR oligomer spectra display a high degree of similarity when compared to the fibril spectra, indicating a high structural similarity of the two aggregation states. Based on comparison to the native state NMR chemical shifts, we suggest that fibril formation via domain-swapping seems unlikely. Moreover, we used our results to test the quality of different amyloid prediction algorithms.


Dissolving sodium hydrosulfide in drinking water is not a good source of hydrogen sulfide for animal studies.
Hydrogen sulfide (H2S) has many physiological and pathological roles in the human body. Sodium hydrosulfide (NaHS) is widely used as a pharmacological tool for assessing H2S effects in biological experiments. Although H2S loss from NaHS solution is a matter of minutes, some animal studies use NaHS in solution as an H2S-donating compound in drinking water. This study addresses whether 30 μM NaHS in drinking water prepared in rat/mouse water bottles remains stable for at least 12-24 h, as presumed by some authors. NaHS solutions (30 μM) were prepared in drinking water and immediately transferred to rat/mice water bottles. Samples were obtained from the tip of water bottles and from inside of the bottles at 0, 1, 2, 3, 4, 5, 6, 12, and 24 h for sulfide measurement using the methylene blue method. Furthermore, NaHS (30 μM) was administered to male and female rats for two weeks, and serum sulfide concentrations were measured every other day in the first week and at the end of the second week. NaHS solution was unstable in the samples obtained from the tip of water bottles; it declined by 72% and 75% after 12 and 24 h, respectively. In the samples obtained from the inside of the water bottles, the decline in the NaHS was not significant until 2 h; however, it decreased by 47% and 72% after 12 and 24 h, respectively. NaHS administration did not affect serum sulfide levels in male and female rats. In conclusion, NaHS solution prepared in drinking water can not be used for H2S donation as the solution is unstable. This route of administration exposes animals to variable and lower-than-expected amounts of NaHS.


Inverse association of N-terminal pro-B-type natriuretic peptide with metabolic syndrome in patients with congestive heart failure.
Metabolic syndrome has been shown to be associated with lower levels of plasma N-terminal pro-B-type natriuretic peptide (Nt-proBNP) in the general population. We sought to elucidate the relationship between Nt-proBNP and components of metabolic syndrome in patients with congestive heart failure (CHF). Fasting blood samples were obtained from 93 patients in our institution. Plasma levels of Nt-proBNP and other biochemical data were measured. The New York Heart Association (NYHA) classification system (I-IV) was used to define the functional capacity of CHF. Metabolic syndrome and its components were defined using diagnostic criteria from the International Diabetes Federation. Forty-nine patients (52.7%) had CHF. There was a positive correlation between plasma Nt-proBNP levels and NYHA functional capacity in CHF patients. Plasma Nt-proBNP levels increased significantly with each increasing NYHA class of the disease. The prevalence of metabolic syndrome in CHF patients was higher than that in patients without CHF. Most importantly, we found that plasma Nt-proBNP levels were lower in CHF patients with metabolic syndrome attributable to inverse relationships between plasma Nt-proBNP and body mass index (β = -0.297), plasma triglyceride (β = -0.286) and homeostasis model assessment of insulin resistance (HOMA-IR; β = -0.346). Fasting glucose to insulin ratio (FGIR, an insulin sensitivity index) was positively associated with plasma Nt-proBNP levels (β = 0.491), and was the independent predictor of plasma Nt-proBNP levels in CHF patients. Plasma Nt-proBNP levels are inversely associated with metabolic syndrome in CHF patients. Reduced plasma Nt-proBNP levels in CHF patients may lead to impaired lipolysis and metabolic function, and may contribute to the development of metabolic syndrome in CHF patients.


Cisplatin induces autophagy to enhance hepatitis B virus replication via activation of ROS/JNK and inhibition of the Akt/mTOR pathway.
Chronic hepatitis B virus (HBV) infection remains a serious global health concern. Cisplatin is a chemotherapeutic agent commonly used to treat various cancers. However, HBV-infected patients receiving chemotherapy are at risk of HBV reactivation via unknown mechanisms, which we aimed to elucidate in this study. We found that autophagy plays a central role in cisplatin-induced HBV replication. Cisplatin treatment induced autophagy in both HBV-replicating cells and an HBV-transgenic mouse model as evident from marked upregulation of microtubule-associated protein 1 light chain 3 (LC3)-II and the accumulation of red fluorescent protein (RFP)-LC3 puncta. Cisplatin induced complete autophagic flux, which was detected via monitoring of p62 degradation and RFP-GFP-LC3 expression. Inhibition of autophagy by chloroquine, 3-methyladenine, or Atg5 knockdown significantly attenuated cisplatin-induced HBV replication. Additionally, cisplatin-induced autophagy could be significantly attenuated by using the ROS scavenger N-acetyl-l-cysteine. Mechanically, cisplatin promoted HBV replication and autophagy through ROS/JNK and AKT/mTOR signaling. Inhibition of JNK or activation of Akt/mTOR signaling reversed cisplatin-mediated autophagy and HBV replication promotion. In contrast, suppression of Akt/mTOR signaling further promoted cisplatin-induced HBV replication. Finally, pharmacotherapeutic inhibition of autophagy or ROS production impaired HBV production induced by cisplatin in vivo. Together, our results indicate that ROS/JNK and mTOR/AKT-mediated autophagy plays an important role in cisplatin-induced HBV reactivation.


Comparison of kidney function between donation after cardiac death and donation after brain death kidney transplantation.
Kidney graft survival is comparable between donation after cardiac death (DCD) and donation after brain death (DBD) kidney transplantation. However, data concerning kidney function after DCD kidney transplantation are lacking. We retrospectively compared kidney function between 64 DCD and 248 DBD kidney transplant recipients. Graft function was assessed using iothalamate glomerular filtration rate at 1, 4, and 12 months, then annually. The primary endpoint was the composite of death-censored graft loss or two consecutive iothalamate glomerular filtration rates less than 50 mL/min/1.73 m² occurring within 5 years from transplantation. Secondary endpoints included death and graft loss or death. Of the 312 patients, 102 (33%) experienced the primary endpoint, 78 (25%) experienced graft loss or death, and 44 (14%) died. In multivariable Cox regression analysis, there was no difference between DCD and DBD recipients regarding the primary endpoint (relative risk [RR], 1.16; P=0.59), death (RR, 0.97; P=0.94), or graft loss or death (RR, 1.09; P=0.79). In the subgroup of 64 DCD recipients, each 10-year increase in donor age was associated with increased risk of the primary endpoint (RR, 1.51; P=0.027) with the highest risk observed for donors older than 45 years (RR, 4.81; P=0.001). Delayed graft function affected 45% of the DCD recipients but had no impact on kidney function, graft survival, or patient survival. Posttransplantation kidney function is comparable between DCD and DBD kidney transplantations. In the subgroup of DCD recipients, kidneys from donors older than 45 years may be associated with a higher risk of poor kidney function; however, this finding requires validation in a larger patient group.


Phenological changes in the southern hemisphere.
Current evidence of phenological responses to recent climate change is substantially biased towards northern hemisphere temperate regions. Given regional differences in climate change, shifts in phenology will not be uniform across the globe, and conclusions drawn from temperate systems in the northern hemisphere might not be applicable to other regions on the planet. We conduct the largest meta-analysis to date of phenological drivers and trends among southern hemisphere species, assessing 1208 long-term datasets from 89 studies on 347 species. Data were mostly from Australasia (Australia and New Zealand), South America and the Antarctic/subantarctic, and focused primarily on plants and birds. This meta-analysis shows an advance in the timing of spring events (with a strong Australian data bias), although substantial differences in trends were apparent among taxonomic groups and regions. When only statistically significant trends were considered, 82% of terrestrial datasets and 42% of marine datasets demonstrated an advance in phenology. Temperature was most frequently identified as the primary driver of phenological changes; however, in many studies it was the only climate variable considered. When precipitation was examined, it often played a key role but, in contrast with temperature, the direction of phenological shifts in response to precipitation variation was difficult to predict a priori. We discuss how phenological information can inform the adaptive capacity of species, their resilience, and constraints on autonomous adaptation. We also highlight serious weaknesses in past and current data collection and analyses at large regional scales (with very few studies in the tropics or from Africa) and dramatic taxonomic biases. If accurate predictions regarding the general effects of climate change on the biology of organisms are to be made, data collection policies focussing on targeting data-deficient regions and taxa need to be financially and logistically supported.


The relationship between the plasma concentration of irbesartan and the antihypertensive response is disclosed by an angiotensin II type 1 receptor polymorphism: results from the Swedish Irbesartan Left Ventricular Hypertrophy Investigation vs. Atenolol (SILVHIA) Trial.
The aim of this study was to investigate the effect of the plasma concentration of irbesartan, a specific angiotensin II type 1 receptor (AT1R) antagonist, and the blood pressure response in relation to AT1R gene polymorphisms. Plasma irbesartan was analyzed in 42 patients with mild-to-moderate hypertension and left ventricular hypertrophy from the Swedish Irbesartan Left Ventricular Hypertrophy Investigation vs. Atenolol (SILVHIA) trial, who were treated with irbesartan as monotherapy for 12 weeks. Blood pressure and irbesartan concentration were measured at trough, i.e., 24 +/- 3 h after the last dose. Five AT1R gene polymorphisms were analyzed by minisequencing. Neither the plasma concentration of irbesartan, nor any of the AT1R polymorphisms were associated with the blood pressure response to irbesartan treatment. However, the interaction term between the plasma concentration of irbesartan and the AT1R C5245T polymorphism was related to the reduction in systolic blood pressure after 12 weeks of treatment (P = 0.025). Furthermore, the plasma concentration of irbesartan was related to the change in systolic blood pressure in individuals homozygous for the AT1R 5245 T allele (r = -0.56, P = 0.030), but not for other genotypes. There was an association between plasma concentrations of irbesartan and the blood pressure response for hypertensive patients with AT1R 5245 TT. Because of the small sample size, this study needs to be viewed as hypothesis generating. This is the first study, to our knowledge, indicating that the concentration-response relationship of an antihypertensive drug may be genotype dependent.


Improved KIR gene and HLA-C KIR ligand sequence-specific primer polymerase chain reaction genotyping using whole genome amplification.
Molecular analysis of genetic polymorphism for clinical or research purposes may be compromised by genomic DNA of limited quality and quantity. In this study, we have successfully tested the feasibility of using whole genome amplification (WGA) to allow genotyping for killer cell immunoglobulin-like receptor (KIR) genes and human leucocyte antigen (HLA)-C KIR ligand dimorphism on HLA-C. WGA was achieved by multiple displacement amplification (MDA) using bacteriophage phi29 polymerase. For KIR genotyping, a revised sequence-specific primer polymerase chain reaction protocol consisting of 23 primer pairs was used avoiding hitherto undetected cross-priming involving KIR2DL1, KIR2DS1, KIR3DL1 and KIR3DS1 alleles. Similarly, MDA-amplified genomic DNA was analyzed for the detection of the HLA-C KIR ligand groups C1 and C2, based on the amino acid K/N dimorphism in position 80.


Experiences of breastfeeding during COVID-19: Lessons for future practical and emotional support.
The COVID-19 pandemic and subsequent lockdown and social distancing led to changes to breastfeeding support available to women in the United Kingdom. Face-to-face professional support was reduced, and face-to-face peer support was cancelled. Anecdotal media accounts highlighted practices separating some mothers and babies in hospitals, alongside inaccurate stories of the safety of breastfeeding circulating. Meanwhile, new families were confined to their homes, separated from families and support networks. Given that we know breastfeeding is best supported by practices that keep mother and baby together, high-quality professional and peer-to-peer support, and positive maternal well-being, it is important to understand the impact of the pandemic upon the ability to breastfeed. To explore this, we conducted an online survey with 1219 breastfeeding mothers in the United Kingdom with a baby 0-12 months old to understand the impact of the pandemic upon breastfeeding duration, experiences and support. The results highlighted two very different experiences: 41.8% of mothers felt that breastfeeding was protected due to lockdown, but 27.0% of mothers struggled to get support and had numerous barriers stemming from lockdown with some stopped breastfeeding before they were ready. Mothers with a lower education, with more challenging living circumstances and from Black and minority ethnic backgrounds were more likely to find the impact of lockdown challenging and stop breastfeeding. The findings are vital in understanding how we now support those women who may be grieving their loss of breastfeeding and are affected by their negative experiences and how we can learn from those with a positive experience to make sure all breastfeeding women are better supported if similar future events arise.


In vitro anticariogenic effects of Drymocallis rupestris extracts and their quality evaluation by HPLC-DAD-MS³ analysis.
In this study, for the first time, we investigated in vitro inhibitory effects of Drymocallis rupestris extracts and their subfractions obtained with solvents of different polarity (aqueous, 50% ethanolic, diethyl ether, ethyl acetate and n-butanolic) against bacterial viability and caries virulence factors of Streptococcus spp. strains. The diethyl ether subfraction (PRU2) showed bacteriostatic and bactericidal activity against mutans streptococci, with minimum inhibitory concentrations (MICs) in the range of 0.75-1.5 mg/mL and minimum bactericidal concentrations (MBCs) in the range of 1.5-3 mg/mL. Furthermore, PRU2 inhibited biofilm formation by Streptococci in a dose-dependent manner. It was also found that all five D. rupestris preparations exhibited diverse inhibitory effects on de novo synthesis of water-insoluble and water-soluble α-D-glucans by glucosyltransferases of the mutans group streptococci. The phytochemical profile of investigated samples was determined by spectrophotometric and chromatographic (HPLC-DAD-MS³) methods. The high polyphenol (total phenol, phenolic acids, tannins, proantocyanidins, and flavonoids) contents were found which correlated with anticariogenic activity of the analyzed samples. The results demonstrate that D. rupestris extracts and their subfractions could become useful supplements for pharmaceutical products as a new anticariogenic agent in a wide range of oral care products. Further studies are necessary to clarify which phytoconstituents of D. rupestris are responsible for anticaries properties.


Influence of Mechanical and Chemical Degradation in the Surface Roughness, Gloss, and Color of Microhybrid Composites.
The aim of this study was to investigate the association of different degradations on the roughness, gloss, and color changes of microhybrid composites. Ten specimens were prepared for Charisma, Amelogen Plus, Point 4, and Opallis resins. Surfaces were polished and baseline measurements of roughness, gloss, and color were recorded. Specimens were then submitted to chemical and mechanical challenges, and the specimens were reevaluated. Roughness and gloss were analyzed by Kruskal -Wallis and Dunn's test (p < 0.05). Color change (ΔE) was analyzed by one-way analysis of variance and Tukey's tests (p < 0.05). The initial and final data were compared using the Wilcoxon test (p < 0.05). Spearman test checked the correlation between the roughness and gloss (p < 0.05). Regarding surface roughness and gloss, there was no difference between composites before challenges. However, all composites showed a significant increase of roughness after challenges, with highest values for Charisma. The gloss was influenced by challenges, evidencing the best gloss for Point 4. Charisma showed the highest value of color change. There was no correlation between surface roughness and gloss for the initial analysis, and after the challenges. Composites were influenced by association of challenges, and Charisma showed the highest changes for roughness, gloss, and color. The type of composite resin influenced the properties of materials, which are surface roughness, gloss, and color change. The dentist should be aware of the performance of different brands, to choose the correct required composite resin for each type of patient or region to be restored.


Leber's Hereditary Optic Neuropathy (LHON): Clinical Experience and Outcomes after Long-Term Idebenone Treatment.
Leber's hereditary optic neuropathy (LHON) is a rare disease. Large studies are difficult to conduct; therefore, case reports provide valuable data. Since 2015, patients have been treated with Idebenone. The aim of this paper is to share our experience with diagnosing and managing patients in different stages of LHON. We designed a case series study, including four patients undergoing genetic testing and ophthalmologic examination. Criteria for Idebenone administration and follow-up were presented. All patients had mutation 11778G>A in MT-ND4. The first patient, an 82-year-old man, with long history of vision loss, had no indication for Idebenone. Two additional cases emerged within the same family: a 40-year-old brother and a 31-year-old sister. Both received Idebenone, with good outcomes only for the female. After a one-year regimen, they were lost to follow-up. The fourth patient, a 46-year-old man, was diagnosed in the subacute stage. Idebenone administration was deferred, allowing progression of visual field defects. After 17 months of treatment, visual improvement appeared. The treatment was continued for 36 months, with short interruptions, resulting in good outcomes. Our study demonstrated positive results with long-term Idebenone use. Contrary to medical literature, our female patient had a favorable evolution, despite the delayed diagnosis.


Asking Children to "Be Helpers" Can Backfire After Setbacks.
Describing behaviors as reflecting categories (e.g., asking children to "be helpers") has been found to increase pro-social behavior. The present studies (N = 139, ages 4-5) tested whether such effects backfire if children experience setbacks while performing category-relevant actions. In Study 1, children were asked either to "be helpers" or "to help," and then pretended to complete a series of successful scenarios (e.g., pouring milk) and unsuccessful scenarios (e.g., spilling milk while trying to pour). After the unsuccessful trials, children asked to "be helpers" had more negative attitudes. In Study 2, asking children to "be helpers" impeded children's helping behavior after they experienced difficulties while trying to help. Implications for how category labels shape beliefs and behavior are discussed.


The Quinazoline Otaplimastat (SP-8203) Reduces the Hemorrhagic Transformation and Mortality Aggravated after Delayed rtPA-Induced Thrombolysis in Cerebral Ischemia.
Acute ischemic stroke is the leading cause of morbidity and mortality worldwide. Recombinant tissue plasminogen activator (rtPA) is the only agent clinically approved by FDA for patients with acute ischemic stroke. However, delayed treatment of rtPA (e.g., more than 3 h after stroke onset) exacerbates ischemic brain damage by causing intracerebral hemorrhage and increasing neurotoxicity. In the present study, we investigated whether the neuroprotant otaplimastat reduced delayed rtPA treatment-evoked neurotoxicity in male Sprague Dawley rats subjected to embolic middle cerebral artery occlusion (eMCAO). Otaplimastat reduced cerebral infarct size and edema and improved neurobehavioral deficits. In particular, otaplimastat markedly reduced intracerebral hemorrhagic transformation and mortality triggered by delayed rtPA treatment, consequently extending the therapeutic time window of rtPA. We further found that ischemia-evoked extracellular matrix metalloproteases (MMPs) expression was closely correlated with cerebral hemorrhagic transformation and brain damage. In ischemic conditions, delayed rtPA treatment further increased brain injury via synergistic expression of MMPs in vascular endothelial cells. In oxygen-glucose-deprived endothelial cells, otaplimastat suppressed the activity rather than protein expression of MMPs by restoring the level of tissue inhibitor of metalloproteinase (TIMP) suppressed in ischemia, and consequently reduced vascular permeation. This paper shows that otaplimastat under clinical trials is a new drug which can inhibit stroke on its own and extend the therapeutic time window of rtPA, especially when administered in combination with rtPA.


Objective measurement of compliance with nasal CPAP treatment for obstructive sleep apnoea syndrome.
Compliance with nasal continuous positive airway pressure (CPAP) has become a major concern, since this treatment is efficacious, but constraining. In 46 consecutive obstructive sleep apnoea (OSA) patients, we measured compliance with nasal CPAP by establishing a mean rate of use, with a built-in time counter read at three-month intervals, over a mean follow-up period of 232 +/- 27 days. The mean rate of use in the whole group was 5.14 +/- 0.31 hours per day. The acceptance rate was 90.9-93.2%, showing that patient acceptance is not a limitation in the use of nasal CPAP.


[Percutaneous closure of a femoral arteriovenous fistula with an endovascular stent graft: a case report].
A 25-year-old male patient underwent primary femoral artery repair following a gunshot injury. Postoperative examination showed a thrill and bruit in the right femoral region. Peripheral angiography performed under right femoral Doppler ultrasound guidance revealed an arteriovenous fistula between the right superficial femoral artery and femoral vein. Through selective angiography from the left femoral artery, an endovascular stent was implanted in the right femoral artery. Control contrast injection showed near-total closure of the fistula and Doppler ultrasonography on the following day showed total closure.


Challenges With Implementation of a Respiratory Therapist-Driven Protocol of Spontaneous Breathing Trials in the Pediatric ICU.
Timely ventilator liberation is crucial in the pediatric ICU. In many pediatric ICUs, the decision to initiate weaning is driven by the physician, which may lead to delays in ventilator liberation. The objectives of this quality improvement project were to develop and implement a respiratory therapist (RT)-led protocol for screening for spontaneous breathing trial (SBT) readiness, to test protocol feasibility, and to evaluate its impact on SBT timing. A retrospective chart review was performed on all intubated patients in the pediatric ICU for 18 months prior to protocol institution. An RT-driven protocol was developed and implemented, enabling consistent screening for SBT readiness. When criteria were met, an SBT was initiated after order placement. The difference in the timing of the first SBT between physician-directed screening and the RT-driven protocol was evaluated. A total of 219 subjects were included in this project (128 pre-intervention; 91 intervention). Baseline demographic data, including mortality risk and endotracheal tube size, were similar in both groups. The time of the first SBT (median [25th, 75th percentile]) was not significantly different between the intervention and preintervention groups (39.5 [25.3, 85.2] vs 42.6 [26.4, 81.3], respectively). There was no difference in mechanical ventilation duration, or length of hospital and ICU stay. The odds of being placed on noninvasive respiratory support were higher in the intervention group at 1 h (odds ratio [95% CI]: 2.29 [1.10, 4.78], P = .03) and 12 h (odds ratio 2.53 [1.23, 5.20], P = .01) postextubation, but the odds of re-intubation did not reach statistical significance (odds ratio 2.60 [0.73, 9.27], P = .14). RT adherence with patient screening was 56.4%. An RT-driven protocol was successfully introduced in an academic pediatric ICU. However, it did not impact time of SBT initiation, potentially due to the difficulty in maintaining adherence over time. RT-driven protocols require further study.


Implanted tissue-cages: a critical evaluation of their relevance in measuring tissue concentrations of antibiotics.
In several species of laboratory animals, the concurrent serum and tissue-cage fluid levels of several antibiotics after a single parenteral dose have markedly different profiles. New work has shown that these differences are mainly due to the relatively impermeable granulomatous sheath which develops around the implanted tissue chambers. The concentrations of an antibiotic in tissue-cage fluid are therefore not representative of its concentration in "interstitial fluid" or extravascular fluid in general. Tissue-cage levels are probably more representative of antibiotic levels to be found inside abscesses, in chronic disease situations, or relatively avascular sites in the body.


Muscle enzyme activity in humans: role of substrate availability and training.
To study the effect of nutrient intake (substrate flux) and training on muscle enzyme activities, 36 untrained healthy men adapted for 7 wk to a fat-rich or a carbohydrate-rich diet. Ten of the 18 subjects on each diet completed an endurance training program, and the remaining 8 served as controls. Maximal oxygen uptake was increased (11%) in the trained groups (P < 0.05). Irrespective of training, beta-hydroxyacyl-CoA-dehydrogenase activity in the vastus lateralis muscle was significantly increased by an average of 25% after adaptation to a fat-rich diet and was unchanged after adaptation to a carbohydrate-rich diet. In contrast, irrespective of diet, muscle citrate synthase activity and hexokinase activity were increased (P < 0.05) after adaptation to training by 17 and 18% in the group fed the carbohydrate, rich diet and by 17 and 12% in the group fed the fat-rich diet, respectively, and were unchanged in the two control groups. We suggest that diet can affect muscle enzymatic adaptation, presumably through an effect on the substrate flux.


Physical aspects of a liquid-filled ionization chamber with pulsed polarizing voltage.
The charge collection in a liquid-filled matrix ionization chamber system has been investigated. This system, which is used for megavoltage radiography, is scanned electronically by switching the polarizing voltage. In such a system the charge collection cannot be described by the classical ionization chamber theory since this is only valid for a fixed polarizing voltage. A model is presented for an ideal plane-parallel ionization chamber with switched polarizing voltage that gives a good qualitative description of the physical phenomena in our nonideal system. The new model predicts the amount of charge collected and the signal-to-noise ratio as a function of several parameters such as electrode distance, polarizing voltage, radiation intensity, and some liquid characteristics. An important result is that the quantum noise contribution can be made quite small. This situation occurs in liquids with a low ion mobility and long ion lifetimes by a charge-integrating effect in the liquid. Experiments were performed to test various aspects of the model. A reasonable agreement is found between theoretical and experimental results. The possible use of ultrahigh-mobility liquids is also discussed.


Rift Valley fever virus NSs protein promotes post-transcriptional downregulation of protein kinase PKR and inhibits eIF2alpha phosphorylation.
Rift Valley fever virus (RVFV) (genus Phlebovirus, family Bunyaviridae) is a negative-stranded RNA virus with a tripartite genome. RVFV is transmitted by mosquitoes and causes fever and severe hemorrhagic illness among humans, and fever and high rates of abortions in livestock. A nonstructural RVFV NSs protein inhibits the transcription of host mRNAs, including interferon-beta mRNA, and is a major virulence factor. The present study explored a novel function of the RVFV NSs protein by testing the replication of RVFV lacking the NSs gene in the presence of actinomycin D (ActD) or alpha-amanitin, both of which served as a surrogate of the host mRNA synthesis suppression function of the NSs. In the presence of the host-transcriptional inhibitors, the replication of RVFV lacking the NSs protein, but not that carrying NSs, induced double-stranded RNA-dependent protein kinase (PKR)-mediated eukaryotic initiation factor (eIF)2alpha phosphorylation, leading to the suppression of host and viral protein translation. RVFV NSs promoted post-transcriptional downregulation of PKR early in the course of the infection and suppressed the phosphorylated eIF2alpha accumulation. These data suggested that a combination of RVFV replication and NSs-induced host transcriptional suppression induces PKR-mediated eIF2alpha phosphorylation, while the NSs facilitates efficient viral translation by downregulating PKR and inhibiting PKR-mediated eIF2alpha phosphorylation. Thus, the two distinct functions of the NSs, i.e., the suppression of host transcription, including that of type I interferon mRNAs, and the downregulation of PKR, work together to prevent host innate antiviral functions, allowing efficient replication and survival of RVFV in infected mammalian hosts.


[Evolutive aspects of post-transfusion hepatitis C. Review of 175 cases].
Hepatitis C virus is the main agent responsible for post-transfusion hepatitis. Progression to chronic hepatitis, cirrhosis and hepatocellular carcinoma is very common. The aim of this study was to evaluate the frequency, timing and factors related to progression of hepatitis C. One hundred seventy five patients with chronic post-transfusion hepatitis C were grouped in a cirrhosis group (n = 92) and a non-cirrhosis group (n = 83). The medium time of development to cirrhosis was 11 +/- 6 years. Patients with cirrhosis were older at the time they received transfusion, used more alcohol and had longer times of evolution. The prognosis was worse in the cirrhosis group with a mortality rate of 28.4% and 9.1% of evolution towards hepatocellular carcinoma, comparing with 5.5% and 0% in the non-cirrhosis group respectively. It is shown that post-transfusion hepatitis C is slowly developing progressive disease which progress is much more rapidly in elderly patients and patients with others factors of liver damage.


The pyruvate requirement of some members of the Mycobacterium tuberculosis complex is due to an inactive pyruvate kinase: implications for in vivo growth.
Through examination of one of the fundamental in vitro characteristics of Mycobacterium bovis--its requirement for pyruvate in glycerol medium--we have revealed a lesion in central metabolism that has profound implications for in vivo growth and nutrition. Not only is M. bovis unable to use glycerol as a sole carbon source but the lack of a functioning pyruvate kinase (PK) means that carbohydrates cannot be used to generate energy. This disruption in sugar catabolism is caused by a single nucleotide polymorphism in pykA, the gene which encodes PK, that substitutes glutamic acid residue 220 with an aspartic acid residue. Substitution of this highly conserved amino acid residue renders PK inactive and thus blocks the ATP generating roles of glycolysis and the pentose phosphate pathway. This mutation was found to occur in other members of the M. tuberculosis complex, namely M. microti and M. africanum. With carbohydrates unable to act as carbon sources, the importance of lipids and gluconeogenesis for growth in vivo becomes apparent. Complementation of M. bovis with the pykA gene from M. tuberculosis H37Rv restored growth on glycerol. Additionally, the presence of a functioning PK caused the colony morphology of the complemented strain to change from the characteristic dysgonic growth of M. bovis to eugonic growth, an appearance normally associated with M. tuberculosis. We also suggest that the glycerol-soaked potato slices used for the derivation of the M. bovis bacillus Calmette and Guérin (BCG) vaccine strain selected for an M. bovis PK+ mutant, a finding that explains the alteration in colony morphology noted during the derivation of BCG. In summary, the disruption of a key step in glycolysis divides the M. tuberculosis complex into two groups with distinct carbon source utilization.


Tendril-less regulates tendril formation in pea leaves.
Tendrils are contact-sensitive, filamentous organs that permit climbing plants to tether to their taller neighbors. Tendrilled legume species are grown as field crops, where the tendrils contribute to the physical support of the crop prior to harvest. The homeotic tendril-less (tl) mutation in garden pea (Pisum sativum), identified almost a century ago, transforms tendrils into leaflets. In this study, we used a systematic marker screen of fast neutron-generated tl deletion mutants to identify Tl as a Class I homeodomain leucine zipper (HDZIP) transcription factor. We confirmed the tendril-less phenotype as loss of function by targeting induced local lesions in genomes (TILLING) in garden pea and by analysis of the tendril-less phenotype of the t mutant in sweet pea (Lathyrus odoratus). The conversion of tendrils into leaflets in both mutants demonstrates that the pea tendril is a modified leaflet, inhibited from completing laminar development by Tl. We provide evidence to show that lamina inhibition requires Unifoliata/LEAFY-mediated Tl expression in organs emerging in the distal region of the leaf primordium. Phylogenetic analyses show that Tl is an unusual Class I HDZIP protein and that tendrils evolved either once or twice in Papilionoid legumes. We suggest that tendrils arose in the Fabeae clade of Papilionoid legumes through acquisition of the Tl gene.


Unexpected phenotypic effects of a transgene integration causing a knockout of the endogenous Contactin-5 gene in mice.
Contactins (Cntn1-6) are a family of neuronal membrane proteins expressed in the brain. They are required for establishing cell-to-cell contacts between neurons and for the growth and maturation of the axons. In humans, structural genomic variations in the Contactin genes are implicated in neurodevelopmental disorders. In addition, population genetic studies associate Contactins loci with obesity and hypertension. Cntn5 knockout mice were first described in 2003, but showed no gross physiological or behavioral abnormalities (just minor auditory defects). We report a novel Cntn5 knockout mouse line generated by a random transgene integration as an outcome of pronuclear microinjection. Investigation of the transgene integration site revealed that the 6Kbp transgene construct coding for the human granulocyte-macrophage colony-stimulating factor (hGMCSF) replaced 170 Kbp of the Cntn5 gene, including four exons. Reverse transcription PCR analysis of the Cntn5 transcripts in the wild-type and transgenic mouse lines showed that splicing of the transgene leads to a set of chimeric hGMCSF-Cntn5 transcript variants, none of which encode functional Cntn5 protein due to introduction of stop codons. Although Cntn5 knockout animals displayed no abnormalities in behavior, we noted that they were leaner, with less body mass and fat percentage than wild-type animals. Their cardiovascular parameters (heart rate, blood pressure and blood flow speed) were elevated compared to controls. These findings link Cntn5 deficiency to obesity and hypertension.


Understanding public perception of hydraulic fracturing: a case study in Spain.
Public acceptance is crucial for the implementation of energy technologies. Hydraulic fracturing is a technology widely used in the USA for natural gas production from shale formations, but currently finds strong public opposition worldwide, especially in Europe. Shale gas exploitation and exploration have the potential to significantly reduce import dependency in several countries, including Spain. To better understand public opinion on this issue, this article reports a survey targeting both the entire Spanish population and the inhabitants of the province of Burgos, the location where shale gas exploration permits have already been issued. Results demonstrate that half of the Spanish population opposes shale gas, and this opposition increases in autonomous communities that are closer to possible exploration sites. The results also show that socio-demographic aspects are not strong predictors of opposition. In addition, Burgos' population show different behaviours toward shale gas that demonstrates that proximity and prospect of shale gas development affects opinion. Finally, there is still a great level of unfamiliarity with high volume hydraulic fracturing and shale gas in both populations sampled.


Renal tubular secretion of the alkanesulfonate 2,3-dimercapto-1-propanesulfonate.
The in vivo tubular secretion and metabolism of 2,3-dimercapto-1-propanesulfonate (DMPS) was examined in the chicken by use of the Sperber technique. Infusion of DMPS into the renal portal circulation of the chicken at rates equal to or less than 7.5 mumol X min-1 X kg body wt-1, resulted in a tubular excretion ratio of 0.5 for DMPS with 90% of the infused DMPS excreted in the urine unchanged. Renal tubular secretion accounted for approximately 90% of the total DMPS excreted into the urine during the infusion of DMPS at a rate of 0.75 mumol X min-1 X kg-1. The secretion of DMPS was saturable and was inhibited by p-aminohippurate (PAH), probenecid, and heptanesulfonate. Taurine (2-aminoethanesulfonate), isethionate (2-hydroxyethanesulfonate), and 2-mercaptoethanesulfonate had no effect on the secretion of DMPS or PAH. Renal tubular secretion may explain several pharmacological characteristics previously reported for DMPS, including the selective removal of heavy metals from the kidney.


Resistance to oncolytic myxoma virus therapy in nf1(-/-)/trp53(-/-) syngeneic mouse glioma models is independent of anti-viral type-I interferon.
Despite promising preclinical studies, oncolytic viral therapy for malignant gliomas has resulted in variable, but underwhelming results in clinical evaluations. Of concern are the low levels of tumour infection and viral replication within the tumour. This discrepancy between the laboratory and the clinic could result from the disparity of xenograft versus syngeneic models in determining in vivo viral infection, replication and treatment efficacy. Here we describe a panel of primary mouse glioma lines derived from Nf1 (+/-) Trp53 (+/-) mice in the C57Bl/6J background for use in the preclinical testing of the oncolytic virus Myxoma (MYXV). These lines show a range of susceptibility to MYXV replication in vitro, but all succumb to viral-mediated cell death. Two of these lines orthotopically grafted produced aggressive gliomas. Intracranial injection of MYXV failed to result in sustained viral replication or treatment efficacy, with minimal tumour infection that was completely resolved by 7 days post-infection. We hypothesized that the stromal production of Type-I interferons (IFNα/β) could explain the resistance seen in these models; however, we found that neither the cell lines in vitro nor the tumours in vivo produce any IFNα/β in response to MYXV infection. To confirm IFNα/β did not play a role in this resistance, we ablated the ability of tumours to respond to IFNα/β via IRF9 knockdown, and generated identical results. Our studies demonstrate that these syngeneic cell lines are relevant preclinical models for testing experimental glioma treatments, and show that IFNα/β is not responsible for the MYXV treatment resistance seen in syngeneic glioma models.


The electrically stimulated gracilis neo-anal sphincter.
Many patients with incontinence gain no relief from their symptoms following traditional surgical treatment and by necessity resort to a stoma. A number of attempts have been made to replace damaged or excised sphincters using the gracilis muscle to fashion a neo-anal sphincter. The results of the published series using the technique of unstimulated graciloplasty for the treatment of anal incontinence and in total anorectal reconstruction are reviewed. The results of these series are conflicting. In an effort to improve on these results the electrically stimulated neo-anal sphincter has been developed. The rationale behind its development and its evolution are discussed. The efficacy of the procedure in treating patients with incontinence or as part of total anorectal reconstruction is assessed.


The structure of the iron-binding protein, FutA1, from Synechocystis 6803.
Cyanobacteria account for a significant percentage of aquatic primary productivity even in areas where the concentrations of essential micronutrients are extremely low. To better understand the mechanism of iron selectivity and transport, the structure of the solute binding domain of an ATP binding cassette iron transporter, FutA1, was determined in the presence and absence of iron. The iron ion is bound within the "C-clamp" structure via four tyrosine and one histidine residues. There are extensive interactions between these ligating residues and the rest of the protein such that the conformations of the side chains remain relatively unchanged as the iron is released by the opening of the metal binding cleft. This is in stark contrast to the zinc-binding protein, ZnuA, where the domains of the metal-binding protein remain relatively fixed, whereas the ligating residues rotate out of the binding pocket upon metal release. The rotation of the domains in FutA1 is facilitated by two flexible beta-strands running along the back of the protein that act like a hinge during domain motion. This motion may require relatively little energy since total contact area between the domains is the same whether the protein is in the open or closed conformation. Consistent with the pH dependence of iron binding, the main trigger for iron release is likely the histidine in the iron-binding site. Finally, neither FutA1 nor FutA2 binds iron as a siderophore complex or in the presence of anions, and both preferentially bind ferrous over ferric ions.


17β-estradiol regulates adenosine triphosphate-binding cassette transporters A1 expression via estrogen receptor A to increase macrophage cholesterol efflux.
The liver is the focus of research on the effects of estrogen on cholesterol metabolism. Few studies have investigated the effects of estrogen on macrophages despite the significance of cells in atherosclerosis. The purpose of this study is to examine the effect of estrogen on macrophage cholesterol efflux. Macrophage cholesterol efflux, oil red O staining, RT-qPCR, Western blotting analyses were used to determine cholesterol metabolize and the expressions of adenosine triphosphate (ATP)-binding cassette transporter G1 (ABCG1) and ATP-binding cassette transporter A1 (ABCA1) in J774A.1 cells, and the effect of these treatments was compared to without adding 17β-estradiol (E2). Gain and loss of estrogen receptor alpha (ERα), liver X receptor α (LXRα) were conducted to study interactions between E2, ERα, LXRα and ABCA. Finally, in mice, we validate the relationship between ERα and ABCA1. E2 increases cholesterol efflux from macrophages and decreases the formation of lipid droplets and positively regulates the expression of ABCA1. This suggests that estrogen receptors (ERs) directly regulate ABCA1 translation. We suppressed ERα, which decreased the mRNA and protein expression of ABCA1. At the mRNA level, E2 treatment could partially counteract these phenomena, but not at the protein level. ABCA1 expression decreased after LXRα was inhibited. This suggests that ABCA1 translation is directly regulated by ERα. In the ovariectomized mouse model of ABCA1 protein expression was significantly reduced in the peritoneal macrophages of the ovariectomy (OVX) group. ABCA1 protein expression was greater in the E2+OVX group than in the OVX group. E2 contributes to the positive regulation of ABCA1 expression and promotes cholesterol efflux in macrophages by binding to ERα. The effect is independent of ABCA1 transcription regulation by LXRα.


Cloning and differential expression of QM like protein homologue from tea [Camellia sinensis (L.) O. Kuntze].
The QM like protein gene encodes for ribosomal protein L10, which is implicated in tumor suppression, transcription factor regulation, and ribosome stability in yeast and mammals. Present study describes cloning of a full-length QM cDNA (CsQM) from tea leaves using differential display of mRNA followed by rapid amplification of cDNA ends. Expression of CsQM was studied in leaves of different stages of development and under various external cues. CsQM contained an open reading frame of 651 bases, encoding 216 amino acids. CsQM shared 71-87% and 85-91% identity at nucleotide and amino acid sequences, respectively with QM genes isolated from other plant species. During active-growth period of tea, higher expression was observed in apical buds that decreased gradually with increasing age of the leaf. During dormancy season, the expression of CsQM gene was severely down-regulated in all the leaves studied. CsQM transcript was found to be down regulated in response to drought stress and abscisic acid treatment but up-regulated by gibberellic acid treatment. A positive association of CsQM transcript abundance with active cellular growth suggested its role in plant growth and development.


Effectiveness of Interferon Beta 1a, compared to Interferon Beta 1b and the usual therapeutic regimen to treat adults with moderate to severe COVID-19: structured summary of a study protocol for a randomized controlled trial.
We will investigate the effectiveness of Interferon Beta 1a, compared to Interferon Beta 1b and the usual therapeutic regimen in COVID-19 in patients that have tested positive and are moderately to severely ill. This is a single center, open label, randomized, controlled, parallel group, clinical trial that will be conducted at Loghman Hakim Medical Education Center in conjunction with Shahid Beheshti University of Medical Sciences. Sixty COVID-19 confirmed cases (using the RT-PCR test) will be enrolled in the trial between April 9th to April 14th 2020. Patients will be randomly assigned to the intervention groups or the control group with the following eligibility criteria: ≥ 18 years of age AND (oxygen saturation (SPO2) ≤ 93% OR respiratory rate ≥ 24) AND at least one of the following: Contactless infrared forehead thermometer temperature of ≥37.8, cough, sputum production, nasal discharge, myalgia, headache or fatigue on admission, and time of onset of the symptoms should be acute (Days ≤ 14). Although Hydroxychloroquine will be administered in a single dose, patients with heart problems (prolonged QT or PR intervals, second- or third-degree heart block, and arrhythmias including torsade de pointes) will be excluded. Other exclusion criteria include using drugs with potential interaction with Hydroxychloroquine + Lopinavir/Ritonavir, Interferon-β 1a, Interferon-β 1b, pregnant or lactating women, history of alcohol or drug addiction in the past 5 years, blood ALT/AST levels > 5 times the upper limit of normal on laboratory results and refusal to participate. This study will be undertaken at the Loghman Hakim Hospital, Shahid Beheshti University of Medical Sciences and Health Services. COVID-19 confirmed patients will be randomly assigned to one of three groups, with 20 patients in each. The first group (Arm 1) will receive Hydroxychloroquine + Lopinavir / Ritonavir (Kaletra) + Interferon-β 1a (Recigen), the second group (Arm 2) will be administered Hydroxychloroquine + Lopinavir / Ritonavir (Kaletra) + Interferon-β 1b (Ziferon), and the control group (Arm 3) will be treated by Hydroxychloroquine + Lopinavir / Ritonavir (Kaletra). Time to clinical improvement is our primary outcome measure. This is an improvement of two points on a seven-category ordinal scale (recommended by the World Health Organization: Coronavirus disease (COVID-2019) R&D. Geneva: World Health Organization) or discharge from the hospital, whichever comes first. Secondary outcomes include mortality from the date of randomization until the last day of the study which will be the day all of the patients have had at least one of the following outcomes: 1) Improvement of two points on a seven-category ordinal scale. 2) Discharge from the hospital 3) Death. If any patient dies, we have reached an important secondary outcome. SpO2 Improvement between the last and first day of hospitalization, using pulse-oximetry. Duration of hospitalization from date of randomization until the date of hospital discharge or date of death from any cause, whichever comes first. Incidence of new mechanical ventilation uses from date of randomization until the last day of the study. Please note that we are trying to add further secondary outcomes and this section of the protocol is still evolving. Statistical analysis will be performed by R version 3.6.1 software. We will use Kaplan-Meier to analyze the time to clinical improvement (compared with a log-rank test). Hazard ratios with 95% confidence intervals will be calculated using the Cox proportional-hazards model in crude and adjusted analysis. Eligible patients will be randomly assigned in a 1:1:1 ratio to receive either Interferon Beta 1a, Interferon Beta 1b or standard care only. Patients will be randomly allocated to three therapeutic arms using permuted, block-randomization to balance the number of patients allocated to each group. The permuted block (three or six patients per block) randomization sequence will be generated, using Package 'randomizeR' in R software version 3.6.1. and placed in individual sealed and opaque envelopes by the statistician. The investigator will enroll the patients and only then open envelopes to assign patients to the different treatment groups. This method of allocation concealment will result in minimum selection and confounding biases. The present research is open-label (no masking) of patients and health care professionals who are undertaking outcome assessment of the primary outcome - time to clinical improvement. Of the 60 patients who underwent randomization, 20 patients were assigned to receive Interferon beta-1a, 20 patients were assigned to receive Interferon beta 1b plus standard care and the rest of patients were assigned to receive the standard care alone. Protocol version 1.2.1. Recruitment is finished, the start date of recruitment was on 9th April 2020 and the end date was on 14th April 2020. Last point of data collection will be the last day on which all of the 60 participants have had an outcome of clinical improvement or death, completing the study's follow-up time window. This study was registered with National Institutes of Health Clinical trials (www.clinicaltrials.gov; identification number NCT04343768, registered April 8, 2020 and first available online April 13, 2020). The full protocol is attached as an additional file, accessible from the Trials website (Additional file 1). In the interest in expediting dissemination of this material, the familiar formatting has been eliminated; this Letter serves as a summary of the key elements of the full protocol.


Evaluation of the genus Listonella and reassignment of Listonella damsela (Love et al.) MacDonell and Colwell to the genus Photobacterium as Photobacterium damsela comb. nov. with an emended description.
The genus Listonella, which was recently described on the basis of 5S rRNA sequence data, was found to be of dubious value on the basis of the results of a comparison of a number of taxonomic studies involving members of the Vibrionaceae. The available data suggest that 5S rRNA sequences may be of limited taxonomic use at the intra- and intergeneric levels, at least for apparently recently evolved groups, such as the Vibrionaceae. In this light, we assessed the generic assignment of the species Listonella damsela. Phenotypic characterization of 12 strains of bacteria assigned to L. damsela, including type strain ATCC 33539, revealed a strong resemblance to members of the genus Photobacterium. All of the strains conformed to major characteristics common to all known Photobacterium species. The characteristics of these organisms included the absence of a flagellar sheath and accumulation of poly-beta-hydroxybutyrate during growth on glucose coupled with the inability to utilize DL-beta-hydroxybutyrate as a sole carbon source. On the basis of the phenotypic data, we propose that L. damsela should be reassigned to the genus Photobacterium as Photobacterium damsela comb. nov.


Kinetic modelling and quantification bias in small animal PET studies with [18F]AB5186, a novel 18 kDa translocator protein radiotracer.
Positron Emission Tomography (PET) imaging with selective 18 kDa translocator protein (TSPO) radiotracers has contributed to our understanding on the role of inflammation in disease development and progression. With an increasing number of rodent models of human disease and expansion of the preclinical PET imaging base worldwide, accurate quantification of longitudinal rodent TSPO PET datasets is necessary. This is particularly relevant as TSPO PET quantification relies on invasive blood sampling due to lack of a suitable tissue reference region. Here we investigate the kinetics and quantification bias of a novel TSPO radiotracer [18F]AB5186 in rats using automatic, manual and image derived input functions. [18F]AB5186 was administered intravenously and dynamic PET imaging was acquired over 2 hours. Arterial blood was collected manually to derive a population based input function or using an automatic blood sampler to derive a plasma input function. Manually sampled blood was also used to analyze the [18F]AB5186 radiometabolite profile in plasma and applied to all groups as a population based dataset. Kinetic models were used to estimate distribution volumes (VT) and [18F]AB5186 outcome measure bias was determined. [18F]AB5186 distribution in rats was consistent with TSPO expression and at 2 h post-injection 50% of parent compound was still present in plasma. Population based manual sampling methods and image derived input function (IDIF) underestimated VT by ~50% and 88% compared with automatic blood sampling, respectively. The VT variability was lower when using IDIF versus arterial blood sampling methods and analysis of the Bland-Altman plots showed a good agreement between methods of analysis. Quantification of TSPO PET rodent data using image-derived methods, which are more amenable for longitudinal scanning of small animals, yields outcome measures with reduced variability and good agreement, albeit biased, compared with invasive blood sampling methods.


Germination variation facilitates the evolution of seed dormancy when coupled with seedling competition.
Fluctuating environmental conditions have consequences for the evolution of life histories because they cause fitness variance. This variance can favor risk-spreading strategies, often known as bet-hedging strategies, in which growth or reproduction is spread over time or space, with some costs, but greater certainty of success. An important example is seed dormancy in annual plants, in which some fraction of seed remains dormant at any given germination opportunity with the potential of germinating later when environmental conditions may differ. Previous theory shows that environmental variation is critical for the evolution of dormancy. However, these studies have focused on temporal variation in reproduction, ignoring the strong observed effects of environmental variation on the germination fraction, a major contributor to fitness variance. We ask what effects germination fluctuations have on selection for dormancy by adding germination fluctuations to existing density-independent (d.i.) and density-dependent (d.d.) models of annual plant dynamics, extending previous analyses by including temporally fluctuating germination. Specifically, we ask how germination variance affects selection on the temporal average germination fraction, here used to define dormancy. When present alone, or when independently varying with other fitness components, germination fluctuations do not affect selection for dormancy in the d.i. model, despite generating fitness variance because this variance contribution is not reduced by higher dormancy. Germination fluctuations have strong effects in the d.d. model, favoring dormancy when present either alone or coupled with variation affecting plant growth. This is because germination variation causes seedling density to vary, which causes variable reproduction through variable intraspecific competition. Dormancy is advantaged under variable reproduction because it creates a more convex relationship between population growth and reproduction leading to benefits from nonlinear averaging. Predictive germination, a positive statistical association between germination and growth, weakens selection for dormancy under strong competition and strengthens it when competition is weak. Our results suggest that variable germination is a potential explanation for high levels of dormancy observed in nature, with implications for life-history theory for fluctuating environments.


Hemerythrins in the microaerophilic bacterium Campylobacter jejuni help protect key iron-sulphur cluster enzymes from oxidative damage.
Microaerophilic bacteria are adapted to low oxygen environments, but the mechanisms by which their growth in air is inhibited are not well understood. The citric acid cycle in the microaerophilic pathogen Campylobacter jejuni is potentially vulnerable, as it employs pyruvate and 2-oxoglutarate:acceptor oxidoreductases (Por and Oor), which contain labile (4Fe-4S) centres. Here, we show that both enzymes are rapidly inactivated after exposure of cells to a fully aerobic environment. We investigated the mechanisms that might protect enzyme activity and identify a role for the hemerythrin HerA (Cj0241). A herA mutant exhibits an aerobic growth defect and reduced Por and Oor activities after exposure to 21% (v/v) oxygen. Slow anaerobic recovery of these activities after oxygen damage was observed, but at similar rates in both wild-type and herA strains, suggesting the role of HerA is to prevent Fe-S cluster damage, rather than promote repair. Another hemerythrin (HerB; Cj1224) also plays a protective role. Purified HerA and HerB exhibited optical absorption, ligand binding and resonance Raman spectra typical of μ-oxo-bridged di-iron containing hemerythrins. We conclude that oxygen lability and poor repair of Por and Oor are major contributors to microaerophily in C. jejuni; hemerythrins help prevent enzyme damage microaerobically or during oxygen transients.


[Effect of chronic alcoholic intoxication on the body of rats].
Experiments involving chronic introduction of ethanol (36 per cent of caloric value) as a part of a specially devised liquid ration demonstrated the trainability of rats to sharply decrease, their antistress resistance to significantly decline, especially in rats kept on a low-protein diet. A general tendency toward a reduced activity of acetylcholinesterase in different sectors of the brain was revealed both in animals receiving alcohol against the background of a normal protein allowance and in those kept on a low-protein diet (when compared with the enzyme activity in animals maintained on a ration with a normal of protein).


Tai Chi Chuan: an ancient wisdom on exercise and health promotion.
Tai Chi Chuan (TCC) is a Chinese conditioning exercise and is well known for its slow and graceful movements. Recent investigations have found that TCC is beneficial to cardiorespiratory function, strength, balance, flexibility, microcirculation and psychological profile. The long-term practice of TCC can attenuate the age decline in physical function, and consequently it is a suitable exercise for the middle-aged and elderly individuals. TCC can be prescribed as an alternative exercise programme for selected patients with cardiovascular, orthopaedic, or neurological diseases, and can reduce the risk of falls in elderly individuals. The exercise intensity of TCC depends on training style, posture and duration. Participants can choose to perform a complete set of TCC or selected movements according to their needs. In conclusion, TCC has potential benefits in health promotion, and is appropriate for implementation in the community.


Ant-plant mutualism: a dietary by-product of a tropical ant's macronutrient requirements.
Many arboreal ants depend on myrmecophytic plants for both food and shelter; in return, these ants defend their host plants against herbivores, which are often insects. Ant-plant and other mutualisms do not necessarily involve the exchange of costly rewards or services; they may instead result from by-product benefits, or positive outcomes that do not entail a cost for one or both partners. Here, we examined whether the plant-ant Allomerus octoarticulatus pays a short-term cost to defend their host plants against herbivores, or whether plant defense is a by-product benefit of ant foraging for insect prey. Because the food offered by ant-plants is usually nitrogen-poor, arboreal ants may balance their diets by consuming insect prey or associating with microbial symbionts to acquire nitrogen, potentially shifting the costs and benefits of plant defense for the ant partner. To determine the effect of ant diet on an ant-plant mutualism, we compared the behavior, morphology, fitness, stable isotope signatures, and gaster microbiomes of A. octoarticulatus ants nesting in Cordia nodosa trees maintained for nearly a year with or without insect herbivores. At the end of the experiment, ants from herbivore exclosures preferred protein-rich baits more than ants in the control (i.e., herbivores present) treatment. Furthermore, workers in the control treatment were heavier than in the herbivore-exclusion treatment, and worker mass predicted reproductive output, suggesting that foraging for insect prey directly increased ant colony fitness. The gaster microbiome of ants was not significantly affected by the herbivore exclusion treatment. We conclude that the defensive behavior of some phytoecious ants is a by-product of their need for external protein sources; thus, the consumption of insect herbivores by ants benefits both the ant colony and the host plant.


Culture, perception, and artistic visualization: a comparative study of children's drawings in three Siberian cultural groups.
In a study of three indigenous and non-indigenous cultural groups in northwestern and northeastern Siberia, framed line tests and a landscape drawing task were used to examine the hypotheses that test-based assessments of context sensitivity and independence are correlated with the amount of contextual information contained in drawings, and with the order in which the focal and background objects are drawn. The results supported these hypotheses, and inspection of the regression relationships suggested that the intergroup variations in test performance were likely to result from differences in the attention accorded to contextual information, as revealed by the drawings. Social and environmental explanations for the group differences in context sensitivity are also discussed. The conclusions support the argument that cultural differences in artistic styles and perceptual tests reflect the same underlying perceptual tendencies, and they are consistent with the argument that these tendencies reflect corresponding differences in patterns of social and environmental interaction.


Challenges and opportunities to manage depression during the menopausal transition and beyond.
Women are at a higher risk than men of developing depression and anxiety and such increased risk might be particularly associated with reproductive cycle events. Recent evidence suggests that the transition to menopause may constitute a window of vulnerability for some women for the development of new onset and recurrent depression. Several biological and environmental factors seem to be independent predictors or modulating factors for the occurrence of depression in menopausal women; they include the presence and severity of hot flushes, sleep disturbances, history of severe premenstrual syndrome or postpartum blues, stressful life events, history of depression, socioeconomic status, and use of hormones and psychotropic agents. The regulation of monoaminergic systems by ovarian hormones might explain, at least in part, the emergence of depressive symptoms and/or anxiety in biologically predisposed subpopulations. The use of transdermal estradiol, as well as serotonergic and noradrenergic antidepressants, is an efficacious strategy in the treatment of depression and vasomotor symptoms in symptomatic women in midlife. In this review, the authors discuss the existing evidence of a greater risk for the development of depression during the menopausal transition and the putative underlying mechanisms contributing to this window of vulnerability. Hormonal and nonhormonal treatment strategies for depression and anxiety in this particular population are critically examined, although more tailored treatment options are still needed.


Climate cycles and forecasts of cutaneous leishmaniasis, a nonstationary vector-borne disease.
Cutaneous leishmaniasis (CL) is one of the main emergent diseases in the Americas. As in other vector-transmitted diseases, its transmission is sensitive to the physical environment, but no study has addressed the nonstationary nature of such relationships or the interannual patterns of cycling of the disease. We studied monthly data, spanning from 1991 to 2001, of CL incidence in Costa Rica using several approaches for nonstationary time series analysis in order to ensure robustness in the description of CL's cycles. Interannual cycles of the disease and the association of these cycles to climate variables were described using frequency and time-frequency techniques for time series analysis. We fitted linear models to the data using climatic predictors, and tested forecasting accuracy for several intervals of time. Forecasts were evaluated using "out of fit" data (i.e., data not used to fit the models). We showed that CL has cycles of approximately 3 y that are coherent with those of temperature and El Niño Southern Oscillation indices (Sea Surface Temperature 4 and Multivariate ENSO Index). Linear models using temperature and MEI can predict satisfactorily CL incidence dynamics up to 12 mo ahead, with an accuracy that varies from 72% to 77% depending on prediction time. They clearly outperform simpler models with no climate predictors, a finding that further supports a dynamical link between the disease and climate.


Stimulation of electron transport as potential novel therapy in Parkinson's disease with mitochondrial dysfunction.
Parkinson's disease (PD) is a neurodegenerative motor disorder characterized by the loss of dopaminergic neurons. This loss of dopaminergic neurons is the pathological hallmark of the disease that results in the characteristic motor syndrome. Restoration of dopamine levels is the basis of current therapy; however, this does not tackle the cause of the disease. While the aetiology of PD remains mostly elusive, mitochondrial dysfunction has been linked to (at least) part of the PD cases. In this review we discuss recent findings in Drosophila melanogaster showing that stimulation of the electron transport chain is beneficial for PD fly models showing Complex I defects and discuss the possible clinical applications of these findings.


Streptococcus pneumoniae clonal complex 199: genetic diversity and tissue-specific virulence.
Streptococcus pneumoniae is an important cause of otitis media and invasive disease. Since introduction of the heptavalent pneumococcal conjugate vaccine, there has been an increase in replacement disease due to serotype 19A clonal complex (CC)199 isolates. The goals of this study were to 1) describe genetic diversity among nineteen CC199 isolates from carriage, middle ear, blood, and cerebrospinal fluid, 2) compare CC199 19A (n = 3) and 15B/C (n = 2) isolates in the chinchilla model for pneumococcal disease, and 3) identify accessory genes associated with tissue-specific disease among a larger collection of S. pneumoniae isolates. CC199 isolates were analyzed by comparative genome hybridization. One hundred and twenty-seven genes were variably present. The CC199 phylogeny split into two main clades, one comprised predominantly of carriage isolates and another of disease isolates. Ability to colonize and cause disease did not differ by serotype in the chinchilla model. However, isolates from the disease clade were associated with faster time to bacteremia compared to carriage clade isolates. One 19A isolate exhibited hypervirulence. Twelve tissue-specific genes/regions were identified by correspondence analysis. After screening a diverse collection of 326 isolates, spr0282 was associated with carriage. Four genes/regions, SP0163, SP0463, SPN05002 and RD8a were associated with middle ear isolates. SPN05002 also associated with blood and CSF, while RD8a associated with blood isolates. The hypervirulent isolate's genome was sequenced using the Solexa paired-end sequencing platform and compared to that of a reference serotype 19A isolate, revealing the presence of a novel 20 kb region with sequence similarity to bacteriophage genes. Genetic factors other than serotype may modulate virulence potential in CC199. These studies have implications for the long-term effectiveness of conjugate vaccines. Ideally, future vaccines would target common proteins to effectively reduce carriage and disease in the vaccinated population.


Bovine herpesvirus-1 gE protein inhibits IFN-β production to enhance replication by promoting MAVS ubiquitination and interfering with the interaction between IRF3 and CBP/p300.
Bovine herpesvirus-1 (BoHV-1) can infect all breeds of cattle and cause respiratory and genital tract diseases. In the process of viral infection, viruses can use their own proteins to suppress the innate immunity of the host and promote its replication; however, the mechanism by which BoHV-1 evades the innate immune response is not fully understood. In this study, we found that rabbits inoculated with the live gene deletion vaccine BoHV-1-△gI/gE/TK generated higher interferon-β (IFN-β) production in the serum, liver, lung and kidney than rabbits inoculated with wt BoHV-1, which led to milder lesions in the lung and kidney. We performed gene deletion and ectopic expression experiments on viral proteins and found that gE was the major protein that inhibited IFN-β expression. Further studies showed that MAVS and IRF3 were the targets of gE, and the specific mechanism was that gE inhibited IFN-β production by promoting MAVS ubiquitination and interfering with the interaction between IRF3 and CBP/p300. These results suggest a new way of BoHV-1 inhibition of IFN-β production to evade the host innate immunity.


[The physician manager--building a clinical leadership in community medicine--the Maccabi Healthcare Services (MHS) model].
Community medicine around the world is facing constant trends of changes. The need to overcome the huge burden of chronic diseases, the need to prioritize and adapt new technologies, and above all, the fact that all these must be done within a given, restricted budget, calls for advanced medical management. In this review we focused on the development of the role of the physician manager in Maccabi Healthcare Services (MHS) over the last 60 years. From what was once a reactive, utilization control-oriented administrative physician role, there has emerged a proactive, formally educated, health quality leader who is expected to lead his clinical colleagues towards achieving the organization's goals. Every organization should answer 4 basic questions in order to encourage/develop the new generation of physician managers. 1. Who am I?--What is the profile and what are the tasks of the physician manager's role? 2. What is the time allocation allotted to the physician manager by the organization to enable him to do his job? 3. What are the educational and managerial tools provided for the 'new" physician manager? 4. What are the rewards that the organization grants to its best people? By addressing the above questions MHS has successfully developed new generations of young clinical leaders who can help MHS management conduct a real dialogue with its clinical physicians in order to maximize the services that our beneficiaries are receiving from the HMO. Our conclusion is that choosing the right people, providing them with the right tools and positioning the physician manager appropriately in the organization's hierarchy will enable the medical care delivery system in Israel to achieve the level of clinical leadership that can lead us towards a better future.


Chromatographic, Chemometric and Antioxidant Assessment of the Equivalence of Granules and Herbal Materials of Angelicae sinensis Radix.
Background: Granules are a popular way of administrating herbal decoctions. However, there are no standardised quality control methods for granules, with few studies comparing the granules to traditional herbal decoctions. This study developed a multi-analytical platform to compare the quality of granule products to herb/decoction pieces of Angelicae Sinensis Radix (Danggui). Methods: A validated ultra-performance liquid chromatography coupled with photodiode array detector (UPLC-PDA) method quantitatively compared the aqueous extracts. Hierarchical agglomerative clustering analysis (HCA) and principal component analysis (PCA) clustered the samples according to three chemical compounds: ferulic acid, caffeic acid and Z-ligustilide. Ferric ion-reducing antioxidant power (FRAP) and 2,2-Diphenyl-1-picrylhydrazyl radical scavenging capacity (DPPH) assessed the antioxidant activity of the samples. Results: HCA and PCA allocated the samples into two main groups: granule products and herb/decoction pieces. Greater differentiation between the samples was obtained with three chemical markers compared to using one marker. The herb/decoction pieces group showed comparatively higher extraction yields and significantly higher DPPH and FRAP (p < 0.05), which was positively correlated to caffeic acid and ferulic acid, respectively. Conclusions: The results confirm the need for the quality assessment of granule products using more than one chemical marker for widespread practitioner and consumer use.


Trifluoromethylation of Electron-Rich Alkenyl Iodides with Fluoroform-Derived "Ligandless" CuCF3.
We herein present a flexible approach for the incorporation of CF3 units into a predefined site of electron-rich alkenes that exploits the regiocontrolled introduction of an iodine handle and subsequent trifluoromethylation of the C(sp2)-I bond using fluoroform-derived "ligandless" CuCF3. The broad substrate scope and functional group tolerance together with the scalability and purity of the resulting products enabled the controlled, late-stage synthesis of single regioisomers of complex CF3-scaffolds, such as sugars, nucleosides (antivirals), and heterocycles (indoles and chromones), with potential for academic and industrial applications.


Toward the virtual cell: automated approaches to building models of subcellular organization "learned" from microscopy images.
We review state-of-the-art computational methods for constructing, from image data, generative statistical models of cellular and nuclear shapes and the arrangement of subcellular structures and proteins within them. These automated approaches allow consistent analysis of images of cells for the purposes of learning the range of possible phenotypes, discriminating between them, and informing further investigation. Such models can also provide realistic geometry and initial protein locations to simulations in order to better understand cellular and subcellular processes. To determine the structures of cellular components and how proteins and other molecules are distributed among them, the generative modeling approach described here can be coupled with high throughput imaging technology to infer and represent subcellular organization from data with few a priori assumptions. We also discuss potential improvements to these methods and future directions for research.


Localization of DNA methyltransferase-1 during oocyte differentiation, in vitro maturation and early embryonic development in cow.
DNA methyltransferase-1 (Dnmt1) is involved in the maintenance of DNA methylation patterns and is crucial for normal mammalian development. The aim of the present study was to assess the localization of Dnmt1 in cow, during the latest phases of oocyte differentiation and during the early stages of segmentation. Dnmt1 expression and localization were assessed in oocytes according to the chromatin configuration, which in turn provides an important epigenetic mechanism for the control of global gene expression and represents a morphological marker of oocyte differentiation. We found that the initial chromatin condensation was accompanied by a slight increase in the level of global DNA methylation, as assessed by 5-methyl-cytosine immunostaining followed by laser scanning confocal microscopy analysis (LSCM). RT-PCR confirmed the presence of Dnmt1 transcripts throughout this phase of oocyte differentiation. Analogously, Dnmt1 immunodetection and LSCM indicated that the protein was always present and localized in the cytoplasm, regardless the chromatin configuration and the level of global DNA methylation. Moreover, our data indicate that while Dnmt1 is retained in the cytoplasm in metaphase II stage oocytes and zygotes, it enters the nuclei of 8-16 cell stage embryos. As suggested in mouse, the functional meaning of the presence of Dnmt1 in the bovine embryo nuclei could be the maintainement of the methylation pattern of imprinted genes. In conclusion, the present work provides useful elements for the study of Dnmt1 function during the late stage of oocyte differentiation, maturation and early embryonic development in mammals.


Regulation of bcl-2 family in hydrogen peroxide-induced apoptosis in human leukemia HL-60 cells.
Numerous types of cells have been shown to undergo apoptosis when exposed to oxidant agent such as hydrogen peroxide. In order to understand the functional relationship between the anti- and pro-apoptotic regulatory proteins in the cells under oxidant stress, we have studied the level of expression of apoptosis regulatory proteins, bcl-2 and bax, in human leukemia HL-60 cells. The exposure of HL-60 cells to different concentrations of H2O2 for 6 h resulted in a typical apoptosis of the cells as characterized by flow cytometry, cell cycle analysis, and DNA fragmantation. There was a block in G1 to S transition and apoptotic cells were mainly derived from S and G2 cells. Kinetic study demonstrated that the levels of both bcl-2-mRNA and -protein expression were decreased with the progression of cellular apoptosis whereas the level of bax-mRNA was unchanged but the expressed bax-protein was not detectable. Cycloheximide, a nonspecific translation inhibitor, did not prevent the hydrogen peroxide-mediated apoptosis in HL-60 cells. These results suggest that the regulation of bcl-2, but not of bax are important factor in the oxidative stress-induced apoptosis in HL-60 cells.


Experiences of sexual relationships of young black women in an atmosphere of coercion.
Negotiations surrounding sexual activity are characterised by multiple power disparities that include race, social status and age, with gender being the most dominant differential in heterosexual interactions. Research has shown that women are physiologically more at risk of contracting HIV than men, as indicated by the higher infection rates of the former. Many African societies operate via a hegemonic masculinity, with patriarchal governance and female subordination being the norm, placing women at even greater risk of HIV infection. In this qualitative phenomenological study, four black school-going adolescent women living in Grahamstown were interviewed using a semi-structured interview to gather data. An interpretative phenomenological analysis was conducted on the data to provide subjective insights of the experiences of the participants with regard to their interactions with men. From the findings, it became apparent that the participants felt pressured, coerced or manipulated by male counterparts. This pressure and coercion was not just felt in their interactions with older men, but also in their romantic partnerships. Three of the participants experienced pressure to engage in sexual intercourse with their boyfriends when they were unwilling or unready, and they reported being faced with additional pressure to engage in unprotected sex. Furthermore, it became apparent that each participant had an underlying fear of being raped and considered this as a genuine threat to her safety and sexual health. The atmosphere within which these participants negotiate their sexual agency is thus heavily informed by male control, coercion and the threat of violence or rape.


Reliability of isokinetic and isometric knee-extensor force in older women.
Because of the need for efficient, consistent strength measurements, the test-retest reliability of concentric, isometric, and eccentric strength; concentric work; and concentric power was determined in older women without a familiarization session. The reliability of measures derived from a single peak score were compared with those derived from an averaged score. On 2 occasions 25 older women with a mean age of 72 +/- 6 years performed 3 submaximal knee extensions and 5 maximal contractions on an isokinetic dynamometer at 90 degrees/s (CON), 0 degrees/s, and -90 degrees/s on both lower limbs. Statistical analyses for peak and averaged values (best 3 contractions of 5) exhibited good relative reliability (ICCs > .88), except for CON power. Typical error as a coefficient of variation and ratio limits of agreement for peak and averaged score values were larger than desired, with CON power scores demonstrating unacceptable error ranges. Although relative reliability of this 1-session assessment protocol was acceptable, further research is needed to determine whether additional practice trials could enhance absolute reliability.


Self-Perceived Confidence of Medical Students Communicating with Pediatric Patients in a 7-Week Pediatric Placement: A Pilot Survey.
Pediatrics is a specialty reserved until later stages of the medical curriculum, with many students receiving early exposure via volunteering opportunities. Self-perceived confidence across the pediatric curriculum is crucial, due to limited clinical exposure before qualification. We aimed to assess the impact of a 7-week pediatric placement on medical students' self-perceived confidence and factors that influenced self-perceived confidence. We conducted a prospective pilot survey on three cohorts of fifth-year students undertaking pediatric placements in 2018. A two-part questionnaire was distributed before and after the placement, evaluating the level of self-confidence in clinical skills using a 10-point scale. Of 103 students, 62 (60%) students completed both questionnaires. Of these, 34 (55%) students reported previous professional experiences with children. There was a significant increase in self-reported confidence scores across ten questions before (mean 5.4 [IQR 4.1-6.1]) and after the placement (7.6 [6.6-8.5], p<0.0001). Subgroup analyses between students with prior professional experience with children and those without revealed a significant difference in preplacement confidence in four pediatric practices: verbal communication, physical engagement, asking sensitive or probing questions, and explaining medical management (p<0.05). There was no significant difference in postplacement confidence between these two groups. Medical students with prior professional experience with children reported higher self-confidence in interacting with pediatric patients prior to placement. However, a large and consistent increase in confidence across the cohort was such that there were no measurable differences at exit. This study supports the value of undergraduate pediatric training in promoting student self-confidence in managing pediatric clinical issues.


Global shifts towards positive species interactions with increasing environmental stress.
The study of positive species interactions is a rapidly evolving field in ecology. Despite decades of research, controversy has emerged as to whether positive and negative interactions predictably shift with increasing environmental stress as hypothesised by the stress-gradient hypothesis (SGH). Here, we provide a synthesis of 727 tests of the SGH in plant communities across the globe to examine its generality across a variety of ecological factors. Our results show that plant interactions change with stress through an outright shift to facilitation (survival) or a reduction in competition (growth and reproduction). In a limited number of cases, plant interactions do not respond to stress, but they never shift towards competition with stress. These findings are consistent across stress types, plant growth forms, life histories, origins (invasive vs. native), climates, ecosystems and methodologies, though the magnitude of the shifts towards facilitation with stress is dependent on these factors. We suggest that future studies should employ standardised definitions and protocols to test the SGH, take a multi-factorial approach that considers variables such as plant traits in addition to stress, and apply the SGH to better understand how species and communities will respond to environmental change.


Modified robust variance estimator for generalized estimating equations with improved small-sample performance.
Generalized estimating equations (GEE (Biometrika 1986; 73(1):13-22) is a general statistical method to fit marginal models for correlated or clustered responses, and it uses a robust sandwich estimator to estimate the variance-covariance matrix of the regression coefficient estimates. While this sandwich estimator is robust to the misspecification of the correlation structure of the responses, its finite sample performance deteriorates as the number of clusters or observations per cluster decreases. To address this limitation, Pan (Biometrika 2001; 88(3):901-906) and Mancl and DeRouen (Biometrics 2001; 57(1):126-134) investigated two modifications to the original sandwich variance estimator. Motivated by the ideas underlying these two modifications, we propose a novel robust variance estimator that combines the strengths of these estimators. Our theoretical and numerical results show that the proposed estimator attains better efficiency and achieves better finite sample performance compared with existing estimators. In particular, when the sample size or cluster size is small, our proposed estimator exhibits lower bias and the resulting confidence intervals for GEE estimates achieve better coverage rates performance. We illustrate the proposed method using data from a dental study.


Functional and evolutionary analysis of host Synaptogyrin-2 in porcine circovirus type 2 susceptibility.
Mammalian evolution has been influenced by viruses for millions of years, leaving signatures of adaptive evolution within genes encoding for viral interacting proteins. Synaptogyrin-2 (SYNGR2) is a transmembrane protein implicated in promoting bacterial and viral infections. A genome-wide association study of pigs experimentally infected with porcine circovirus type 2b (PCV2b) uncovered a missense mutation (SYNGR2 p.Arg63Cys) associated with viral load. In this study, CRISPR/Cas9-mediated gene editing of the porcine kidney 15 (PK15, wtSYNGR2+p.63Arg) cell line generated clones homozygous for the favorable SYNGR2 p.63Cys allele (emSYNGR2+p.63Cys). Infection of edited clones resulted in decreased PCV2 replication compared to wildtype PK15 (P<0.05), with consistent effects across genetically distinct PCV2b and PCV2d isolates. Sequence analyses of wild and domestic pigs (n>700) revealed the favorable SYNGR2 p.63Cys allele is unique to domestic pigs and more predominant in European than Asian breeds. A haplotype defined by the SYNGR2 p.63Cys allele was likely derived from an ancestral haplotype nearly fixed within European (0.977) but absent from Asian wild boar. We hypothesize that the SYNGR2 p.63Cys allele arose post-domestication in ancestral European swine. Decreased genetic diversity in homozygotes for the SYNGR2 p.63Cys allele compared to SYNGR2 p.63Arg, corroborates a rapid increase in frequency of SYGNR2 p.63Cys via positive selection. Signatures of adaptive evolution across mammalian species were also identified within SYNGR2 intraluminal loop domains, coinciding with the location of SYNGR2 p.Arg63Cys. Therefore, SYNGR2 may reflect a novel component of the host-virus evolutionary arms race across mammals with SYNGR2 p.Arg63Cys representing a species-specific example of putative adaptive evolution.