Insertion and excision of the transposable element mariner in Drosophila.
The transposable element mariner is active in both germline and somatic cells of Drosophila mauritiana. Activity of the element is greatly enhanced in the presence of Mos1, a genetic factor identified as an autonomous copy of mariner. A strain of D. mauritiana containing Mos1 and other copies of mariner was used to initiate a screen for visible mutations. More than 20 mutations were obtained, including alleles of white, yellow and vermilion. Six alleles were characterized at the molecular level, and all were found to contain a mariner element inserted into the affected gene. Four insertions into the white locus were sequenced to determine the exact site of insertion of mariner. There appears to be little sequence specificity requirement for mariner insertion, other than an absolute requirement for the dinucleotide TA, which is duplicated upon insertion. Sequences of phenotypically wild-type germline and somatic revertants obtained from various white alleles, including the previously isolated wpch allele, were obtained using the polymerase chain reaction. Mariner excision is imprecise in both germline and soma, and the most frequent excision events are the same in the two tissues. Mutant derivatives of wpch were also studied, and were found to exhibit a wide range of molecular structures and phenotypes.


Intraocular HIV-1-specific IgG synthesis in a patient with CMV retinitis.
A sight-threatening cytomegalovirus (CMV) retinitis is often associated with acquired immune deficiency syndrome (AIDS). We report the detection and quantitation of antibodies specific for CMV and human immunodeficiency virus type 1 (HIV-1) in the vitreous fluid and serum of an AIDS patient suffering from bilateral CMV retinitis. The ratio of the concentrations of HIV-1-specific immunoglobulin G (IgG) to total IgG in the vitreous fluid was found to be higher than that of the peripheral blood, indicating a local production of HIV-1-specific IgG synthesis within the ocular compartment. In contrast, CMV-specific IgG levels in serum were found to be higher than that of vitreous fluid, indicating an intact blood-ocular barrier. CMV and HIV were also isolated from the ocular tissues of this patient. These findings are consistent with the hypothesis that CMV retinitis may be associated with HIV-1 infection of the ocular tissues, which evokes an HIV-1-specific humoral immune response locally within the ocular compartment.


Factors influencing nurses' decisions to activate medical emergency teams.
To evaluate the relationship between nurse demographics and correct identification of clinical situations warranting specific nursing actions, including activation of the medical emergency team. If abnormal physiology is left untreated, the patient may develop cardiac arrest. Nurses in general wards are those who perceive any clinical deterioration in patients. A descriptive, quantitative design was selected. An anonymous survey with 13 multiple choice questions was distributed to 150 randomly selected nurses working in general medical and surgical wards of a large tertiary hospital in Athens, Greece. After explanation of the purposes of the study, 94 nurses (response ratio: 62%) agreed to respond to the questionnaire. Categories with the greatest nursing concern were patients with heart rate<40/minute, an atypical thoracic pain, foreign body airway obstruction and bronchial secretions, respiratory rate<5/minute and heart rate=100/minute. However, almost 50% of nurses were able to accurately identify the critical nursing action for patients with respiratory rate<4/minute, 72% for patients with airway obstruction and 73% for patients with chest pain. Nurses who had graduated from a four-year educational programme identified clinical situations that necessitated medical emergency team activation in a significantly higher rate and also scored significantly higher in questions concerning clinical evaluation than nurses who had graduated from a two-year educational programme. Activation of the medical emergency team is influenced by factors such as level of education and cardiopulmonary resuscitation courses attendance. Graduating from a four-year educational programme helps nurses identify emergencies. However, irrespective of the educational programme they have followed, undertaking a basic life support or advanced life support provider course is critical as it helps them identify cardiac or respiratory emergencies.


The tobacco quitline setting as a teachable moment: The Educating Quitline Users About Lung (EQUAL) cancer screening randomized trial.
Although lung cancer screening (LCS) using low-dose CT is recommended for high-risk individuals, screening adherence remains low. We conducted a randomized trial to compare two methods of providing LCS education to Maryland Tobacco Quitline (MTQ) callers in order to assess whether this setting may serve as a teachable moment for LCS-eligible individuals. MTQ callers (50-80 years, 20+ pack-years, prior LCS ≥12 months) completed the baseline and were randomized to the Print- or Web-based version of ShouldIScreen.com. Participants completed 1- and 4-month follow-up assessments to evaluate intervention engagement and LCS-related outcomes. Participants (Print = 152, Web = 146) were 61.7 (SD = 6.3) years old and reported 63.5 pack-years (SD = 36.0). Most identified as Black (54.2%), female (66.1%), having internet access (78.9%), completing other recommended cancer screenings (86.3%), and that they would undergo LCS if recommended by their provider (91.3%). By 4 months, significantly more Print (75.0%) than Web (61.6%) participants had read the materials (P = .01). Most reported the interventions contained "the right amount" of information (92.6%) and prepared them to talk with their doctor (57.2%). Regarding screening-related outcomes, 42.8% (Print) and 43.8% (Web) had scheduled or completed a low-dose CT scan or a shared decision-making visit (P = .86). In a racially diverse sample of LCS-eligible quitline callers, offering LCS educational materials resulted in high intervention engagement and screening-related appointments. As >20% did not have internet access, providing participants' preferred modality (web/print) may improve intervention engagement and knowledge. Improving LCS awareness represents an important opportunity to increase screening among eligible but unscreened quitline callers.


[Soil seed bank and its correlations with aboveground vegetation and environmental factors in water level fluctuating zone of Danjiangkou Reservoir, Central China].
Taking the water level fluctuating zone of the Danjiangkou Reservoir as a case, and by the method of hierarchical cluster analysis, the soil seed banks at 37 sampling plots within the areas of 140-145 m elevation were divided into 6 groups, and the species composition, density, and diversity of the soil seed banks among the groups were compared. The differences between the soil seed banks and the aboveground vegetations were analyzed by S0rensen similarity coefficient, and the correlations among the soil seed banks, aboveground vegetations, and environmental factors were explored by principal component analysis (PCA) and multivariable regression analysis. At the same altitudes of the water level fluctuating zone, the species composition of the soil seed banks had obvious heterogeneity, and the density and diversity indices of the soil seed banks among different groups were great. The similarity coefficient between the soil seed banks and aboveground vegetations was low, and the species number in the soil seed banks was obviously lesser than that in the aboveground vegetations. The density of the soil seed banks was highly positively correlated with the aboveground vegetations coverage and species number and the soil texture, but highly negatively correlated with the soil water-holding capacity and soil porosity.


Reactive oxygen species and fatigue-induced prolonged low-frequency force depression in skeletal muscle fibres of rats, mice and SOD2 overexpressing mice.
Skeletal muscle often shows a delayed force recovery after fatiguing stimulation, especially at low stimulation frequencies. In this study we focus on the role of reactive oxygen species (ROS) in this fatigue-induced prolonged low-frequency force depression. Intact, single muscle fibres were dissected from flexor digitorum brevis (FDB) muscles of rats and wild-type and superoxide dismutase 2 (SOD2) overexpressing mice. Force and myoplasmic free [Ca(2+)] ([Ca(2+)](i)) were measured. Fibres were stimulated at different frequencies before and 30 min after fatigue induced by repeated tetani. The results show a marked force decrease at low stimulation frequencies 30 min after fatiguing stimulation in all fibres. This decrease was associated with reduced tetanic [Ca(2+)](i) in wild-type mouse fibres, whereas rat fibres and mouse SOD2 overexpressing fibres instead displayed a decreased myofibrillar Ca(2+) sensitivity. The SOD activity was approximately 50% lower in wild-type mouse than in rat FDB muscles. Myoplasmic ROS increased during repeated tetanic stimulation in rat fibres but not in wild-type mouse fibres. The decreased Ca(2+) sensitivity in rat fibres could be partially reversed by application of the reducing agent dithiothreitol, whereas the decrease in tetanic [Ca(2+)](i) in wild-type mouse fibres was not affected by dithiothreitol or the antioxidant N-acetylcysteine. In conclusion, we describe two different causes of fatigue-induced prolonged low-frequency force depression, which correlate to differences in SOD activity and ROS metabolism. These findings may have clinical implications since ROS-mediated impairments in myofibrillar function can be counteracted by reductants and antioxidants, whereas changes in SR Ca(2+) handling appear more resistant to interventions.


Quantitative genetic analysis of biomass and wood chemistry of Populus under different nitrogen levels.
The genetic control of carbon allocation and partitioning in woody perennial plants is poorly understood despite its importance for carbon sequestration, biofuels and other wood-based industries. It is also unclear how environmental cues, such as nitrogen availability, impact the genes that regulate growth, biomass allocation and wood composition in trees. We phenotyped 396 clonally replicated genotypes of an interspecific pseudo-backcross pedigree of Populus for wood composition and biomass traits in above- and below-ground organs. The loci that regulate growth, carbon allocation and partitioning under two nitrogen conditions were identified, defining the contribution of environmental cues to their genetic control. Sixty-three quantitative trait loci were identified for the 20 traits analyzed. The majority of quantitative trait loci are specific to one of the two nitrogen treatments, demonstrating significant nitrogen-dependent genetic control. A highly significant genetic correlation was observed between plant growth and lignin/cellulose composition, and quantitative trait loci co-localization identified the genomic position of potential pleiotropic regulators. Pleiotropic loci linking higher growth rates to wood with less lignin are excellent targets to engineer tree germplasm improved for pulp, paper and cellulosic ethanol production. The causative genes are being identified with a genetical genomics approach.


Plasma adrenomedullin peptides and precursor levels in pulmonary arterial hypertension disease severity and risk stratification.
Adrenomedullin is a potent vasodilatory peptide, linked to pulmonary arterial hypertension pathology. Proximity extension assays were utilized to study plasma biomarkers related to vasoregulation, with focus on adrenomedullin peptides and precursor levels, collectively referred to as ADM. ADM was measured in 48 treatment-naïve pulmonary arterial hypertension patients at diagnosis, and in 31 of them at an early treatment follow-up. Plasma ADM was additionally assessed in patients with chronic thromboembolic pulmonary hypertension (n = 20) and pulmonary hypertension due to heart failure with preserved (HFpEF(PH)) (n = 33) or reduced (HFrEF(PH)) (n = 36) ejection fraction, as well as healthy controls (n = 16). ADM was studied in relation to pulmonary arterial hypertension hemodynamics, risk assessment, prognosis, treatment response, and differentiation. Plasma ADM levels in pulmonary arterial hypertension patients at diagnosis were higher than in healthy controls (p < 0.001), similar as in chronic thromboembolic pulmonary hypertension patients (p = ns), but lower compared to HFpEF(PH) (p < 0.03) and HFrEF(PH) (p < 0.001). In pulmonary arterial hypertension, specifically, plasma ADM at diagnosis correlated mainly to mean right atrial pressure (r = 0.73, p < 0.001), N-terminal prohormone of brain natriuretic peptide (r = 0.75, p < 0.001), six-minute walking distance (r = -0.57, p < 0.001), and venous oxygen saturation (r = -0.57, p < 0.001). ADM also correlated to the ECS/ERS- (r = 0.74, p < 0.001) and REVEAL risk scores (r = 0.54, p < 0.001) at pulmonary arterial hypertension diagnosis. Plasma ADM in pulmonary arterial hypertension patients was unaltered at early treatment follow-up compared to baseline (p = ns). Pulmonary arterial hypertension patients with supra-median ADM at diagnosis showed worse overall survival than those with infra-median levels (median survival 34 versus 66 months, p = 0.0077). In conclusion, the present results suggest that baseline plasma ADM levels mirror disease severity, correlating to both ECS/ERS- and the REVEAL risk scores.


A computational study on plasticity during theta cycles at Schaffer collateral synapses on CA1 pyramidal cells in the hippocampus.
Cellular activity in the CA1 area of the hippocampus waxes and wanes at theta frequency (4-8 Hz) during exploratory behavior of rats. Perisomatic inhibition onto pyramidal cells tends to be strongest out of phase with pyramidal cell activity, whereas dendritic inhibition is strongest in phase with pyramidal cell activity. Synaptic plasticity also varies across the theta cycle, from strong long-term potentiation (LTP) to long-term depression (LTD), putatively corresponding to encoding and retrieval phases for information patterns encoded by pyramidal cell activity (Hasselmo et al. (2002a) Neural Comput 14:793-817). The mechanisms underpinning the phasic changes in plasticity are not clear, but it is likely that inhibition plays a role by affecting levels of electrical activity and calcium concentration at synapses. We explore the properties of synaptic plasticity during theta at Schaffer collateral synapses on CA1 pyramidal neurons and the influence of spatially and temporally targeted inhibition using a detailed multicompartmental model of the CA1 pyramidal neuron microcircuit and a phenomenological model of synaptic plasticity. The results suggest CA3-CA1 synapses are potentiated on one phase of theta due to high calcium levels provided by paired weak CA3 and layer III entorhinal cortex (EC) inputs even when somatic spiking is inhibited by perisomatic interneuron activity. Weak CA3 inputs alone induce lower calcium transients and result in depression of the CA3-CA1 synapses. These synapses are depressed if activated in phase with dendritic inhibition as strong CA3 inputs alone are not able to cause high calcium in this theta phase even though the CA1 pyramidal neuron shows somatic spiking. Dendritic inhibition acts as a switch that prevents LTP and promotes LTD during the retrieval phases of the theta rhythm in CA1 pyramidal cell. This may be important for not overly reinforcing recalled memories and in forgetting no longer relevant memories.


Hyporesponsiveness to lipopolysaccharide alters the composition of NF-kappaB binding to the regulatory regions of inducible nitric oxide synthase gene.
Repeated exposure to bacterial endotoxin causes a diminished response by the host to further exposure. One important feature of this hyporesponsiveness is a reduced macrophage production of nitric oxide (NO) via the inducible nitric oxide synthase (iNOS) pathway. Using a murine macrophage model, we observed that hyporesponsiveness was accompanied by a decrease in the levels of NO release (measured as nitrite), iNOS protein and iNOS gene transcription. The expression of the putative lipopolysaccharide (LPS) receptor, CD14, was not altered. In vivo genomic footprinting showed that the same binding sites are occupied in the iNOS promoter and enhancer of desensitized macrophages and of LPS-responsive macrophages, yet the composition of NF-kappaB in the nuclei of these cells was found to be altered. The transcriptionally inactive homodimer p50-p50 represented the predominant binding activity in nuclei from LPS-pretreated cells before and after stimulation. Nuclei from cells which had not been pretreated but were stimulated contained more of the transcriptionally active p50-p65 heterodimer than their pretreated counterparts. Consistent with this, the cytosolic steady-state level of an inhibitor of NF-kappaB activity, I-kappaBalpha, was decreased in normal cells but not in pretreated cells. We propose that the presence of an overwhelming excess of transcriptionally inactive p50 homodimers on their kappaB sites in the iNOS control region in pretreated cells may block kappaB site binding by p50-p65, thereby reducing the activity of the protein complex governing iNOS transcription.


The use of silver nitrate in wound management.
The use of silver nitrate for chronic wounds such as leg ulcers, as well as burns and acute wounds has already been described in literature. However there is currently conflicting evidence in the literature to support the use of silver nitrate. We present our experience on two cases with extensive co-morbidities associated, where the use of silver nitrate application reduce dramatically the size of large wounds, which eventually healed, avoiding the patients to undergone surgery. Furthermore to reduce pain and distress associated with the application we used local anaesthetic prior to applying the silver nitrate.


Occult Nodal Metastasis in Oral Cavity Cancers.
In squamous cell carcinoma (SCC) of the oral cavity, there is always a risk of occult metastasis to neck nodes in the clinically and radiologically negative neck (N0). Therefore, elective neck dissection (END) has ever been under discussion since the beginning of their routine use for the management of neck for oral carcinomas. The purpose of the current study is to identify the percentage of occult nodal metastasis to neck levels I-V in the cases of oral carcinoma who were treated for the N0 with END. Patients who were treated between June 2005 and May 2010 with END from neck levels I to V for the management of N0 with oral SCC had been identified from the database of Aga Khan University Hospital. Those who met the inclusion and exclusion criteria were included in the study. Data were analyzed using SPSS 16 software. Using descriptive statistics, the mean was computed for the quantitative variable (age). Frequencies and percentages were calculated for gender, site, tumor grade, and lymph node involvement for each neck level. A total of 50 patients were included in the study. There were 38 males and 12 females. The mean age was 47 (range 25-72). The most common site of the tumor was buccal mucosa in 50% of the cases followed by tongue 20%, then floor of mouth 14%, dentoalveolar ridge 8%, retromolar area 4%, lip 2%, and hard palate 2%. Histopathological grading of tumors showed well-differentiated 28%, moderately differentiated 33%, and poorly differentiated 6%. Twenty-seven out of 50 patients were found positive for nodal metastasis on final postoperative histopathology. Neck node metastasis at level I was found in 22 patients, at level II in 16 patients, at level III in seven patients, and at level IV in two patients. The level V was found free of metastasis in all of the cases. The rate of occult metastatic disease to the neck nodes was similar to that found in the literature. Both early and advanced local disease is associated with a risk of occult metastasis. END for neck levels I-V is, therefore, recommended for the management of the N0 in all cases of oral SCCs. Spread to levels IV and V is rare and these levels should not be a part of routine END.


Endodontic access cavities: Fears and difficulties of Moroccan dental students related to teachers, self-confidence and technical protocol.
Fears and difficulties of Moroccan dental students regarding endodontic access cavity were evaluated among 3 items: (i) teachers, (ii) student confidence and (iii) access cavity protocol. A three Likert scale was used to rate 24 statements. Specific scores between 1 and 3 and mean scores between 0 and 1 were used to assess statements and items, respectively. Among 116 students, the mean score of difficulty related to teachers (0.48 ± 0.14) was significantly low (p < 0.001) compared to student confidence (0.69 ± 0.12) and cavity protocol (0.68 ± 0.08). The number of teachers was esteemed insufficient, additional training sessions were requested and maxillary teeth and molars were rated the most difficult with specific scores of 1.91 ± 0.80, 2.72 ± 0.61, 2.22 ± 0.67 and 2.47 ± 0.74, respectively. The highest specific score referred to the risk of making ledges (2.8 ± 0.56) or perforations (2.76 ± 0.59). Students expressed a lack of confidence, difficulty with upper molars and fears of making procedural errors.


[Course of minimal oxygen consumption in the young rat during protein energy malnutrition followed by return to a balanced diet].
Growth and resting oxygen consumption (VO2 minimal) were studied on two strains of six post-weaning male rats during 60 days. The first strain was fed with a balanced diet (15% casein) for 60 days, the second received a 2% casein diet for 30 days, and then the balanced diet for the remaining 30 days. The 2% casein diet completely stopped the body weight and VO2 minimal increases. When return to a balanced diet, these parameters show an evolution similar to that of controls of the same body weight. The VO2 minimal had the same kind of evolution as body weight and was well correlated with growth speed. The protein restriction involves a strong and temporary reduction of the VO2 minimal, which will correspond to a nutritional stress.


Serum cobalt concentrations remain at low levels at a minimum of 20 years following metal-on-metal total hip arthroplasty.
Second-generation metal-on-metal (MoM) articulations in total hip arthroplasty (THA) were introduced in order to reduce wear-related complications. The current study reports on the serum cobalt levels and the clinical outcome at a minimum of 20 years following THA with a MoM (Metasul) or a ceramic-on-polyethylene (CoP) bearing. The present study provides an update of a previously published prospective randomized controlled study, evaluating the serum cobalt levels of a consecutive cohort of 100 patients following THA with a MoM or a CoP articulation. A total of 31 patients were available for clinical and radiological follow-up examination. After exclusion of 11 patients because of other cobalt-containing implants, 20 patients (MoM (n = 11); CoP (n = 9)) with a mean age of 69 years (42 to 97) were analyzed. Serum cobalt levels were compared to serum cobalt levels five years out of surgery. The median cobalt concentration in the MoM group was 1.04 μg/l (interquartile range (IQR) 0.64 to 1.70) at a mean of 21 years (20 to 24) postoperatively and these values were similar (p = 0.799) to cobalt levels at five years. In the CoP control group, the median cobalt levels were below the detection limit (< 0.3 μg/l; median 0.15 μg/l, IQR 0.15 to 0.75) at 20 years. The mean Harris Hip Score was 91.4 points (61 to 100) in the MoM group and 92.8 points (63 to 100) in the CoP group. This study represents the longest follow-up series evaluating the serum cobalt levels after 28 mm head MoM bearing THA and shows that serum cobalt concentrations remain at low levels at a mean of 21 years (20 to 24) after implantation.Cite this article: Bone Joint Res. 2020;9(3):145-150.


99mTc-Pertechnetate Scintigraphy Predicts Successful Postoperative Ablation in Differentiated Thyroid Carcinoma Patients Treated with Low Radioiodine Activities.
Postoperative routine radioiodine (RAI) treatment is currently debated for patients with low-risk differentiated thyroid carcinoma (DTC) patients. If performed, a low ¹³¹I activity (i.e., 1 to 2 GBq) is recommended with the aim to ablate thyroid remnant and facilitate subsequent follow-up by thyroglobulin measurement. The purpose of this study was to evaluate the relationship between postsurgical technetium-99m (99mTc)-pertechnetate scintigraphy and the rate of successful remnant ablation after low activity radioiodine ablation in patients with DTC. Enrolled were 193 patients with low risk DTC who underwent total thyroidectomy and RAI ablation with a fixed 1.1 GBq activity of ¹³¹I. 99mTc-pertechnetate scans were done and thyrotropin stimulated thyroglobulin (sTg) levels measured just before ablation. Ablation effectiveness was assessed 6 to 12 months later by sTg measurement, neck ultrasound and diagnostic whole body scan. A negative 99mTc-perthecnetate scans was the best predictor of successful ablation (P<0.001) followed by preablative sTg levels <0.8 ng/mL (P=0.008) and 99mTc-pertechnetate uptake rate values <0.9% (P=0.065). Neither sex nor age of the patient at the time of ablation or tumor histology and size showed a significant association with the rate of successful ablation. The 99mTc-pertechnetate scintigraphy is a simple and feasible tool to predict effectiveness of low activity ¹³¹I thyroid to ablate thyroid remnants in patients with DTC.


Stunting, underweight and wasting among Integrated Child Development Services (ICDS) scheme children aged 3-5 years of Chapra, Nadia District, West Bengal, India.
This study investigated age and sex variations in height and weight, levels of stunting, underweight and wasting among 533 (254 boys; 279 girls) 3- to 5-year-old rural children of Bengalee ethnicity at 11 Integrated Child Development Services centres of Nadia District, West Bengal, India. Height-for-age, weight-for-age and weight-for-height < -2 z-scores were used to evaluate stunting, underweight and wasting, respectively, following the National Center for Health Statistics (NCHS) Guidelines. Results revealed that boys were significantly heavier than girls at age 3 years. Significant age differences existed in mean height and weight in both sexes. Mean z-scores of height-for-age, weight-for-age and weight-for-height were lower than those of NCHS for both sexes at all ages. The overall (age and sex combined) rates of stunting, underweight and wasting were 23.9%, 31.0% and 9.4%, respectively. The rate of underweight and wasting was higher among girls (underweight = 35.1%, wasting = 12.2%) compared with boys (underweight = 26.5%, wasting = 6.3%). In general, the frequency of stunting increased with increasing age in both sexes. Based on the World Health Organization classification of severity of malnutrition, the overall prevalence of underweight was very high (>or=30%). The prevalence rates of stunting (20-29%) and wasting (5-9%) were medium. In conclusion, the nutritional status of the subjects is unsatisfactory. There is scope for improvement in the form of enhanced supplementary nutrition.


[The burden of neoplasm in Brazil: mortality and hospital morbidity from 2002 to 2004].
To describe mortality and the hospital morbidity by neoplasias in Brazil and regions according to gender. Data of deaths were obtained from the Mortality Information System and of hospital morbidity from the Hospital Information System. Deaths were categorized according to primary tumor sites, selected in accordance with the tenth revision of the International Classification of Diseases. The population data were drawn from the inter census estimates of the IBGE (Brazilian Institute of Geography and Statistics). The period of analysis was the triennial 2002-2004, with the most recent mortality data in Brazil. The average of this period was calculated to ensure greater stability of the rates. Between 2002 and 2004, 405,415 deaths from neoplasias occurred in Brazil. The highest rates of mortality were identified in the South and South-East regions. For men, cancer of the trachea, bronchi and lungs were the malignant neoplasias with the highest mortality rate while for women breast cancer was highest. Breast cancer and cancer of the uterine cervix are those requiring the largest number of in-hospital admissions. In internments, leukemia presented the highest average cost and total cost. The burden of neoplasms is extremely high in Brazil and public policies focused on the population must be given priority for an effective control of mortality and morbidity.


Therapeutic measures after hepatitis B virus infection: postexposure prophylaxis.
Hepatitis B postexposure prophylaxis is here reviewed. Every contact with hepatitis B virus can cause an infection which may be either acute, subclinical, or progressive, the latter potentially leading to chronic liver disease and hepatocellular carcinoma. Direct injection of large quantities of HBsAg-positive blood is almost invariably contagious. While casual person-to-person contact rarely causes disease, the multiplicity of exposure in the hospital environment or the home increases the risk for transmission. Several studies have shown that postexposure prophylaxis using passive immunization with specific hepatitis B antibodies is possible. Hepatitis B immune globulin (HBIG) with a minimum titre of 100 IU/ml should be used. In practice, most preparations contain 500 IU/ml. The use of HBIG post-exposure prophylaxis should be limited to needlestick injury, sexual exposure, and perinatal contact of neonates with HBsAg-positive mothers. Routine vaccination as an adjunct to HBIG administration is recommended. HBIG does not decrease the immunogenic properties of the vaccine provided that the injection is not made at the same site. With increasing use of hepatitis B vaccines, the need for therapeutic intervention will hopefully be considerably diminished. Prevention rather than therapy should be stressed.


Identification of a Tbx1/Tbx2/Tbx3 genetic pathway governing pharyngeal and arterial pole morphogenesis.
The 22q11.2 deletion syndrome (22q11.2DS) is the most common microdeletion disorder and is characterized by abnormal development of the pharyngeal apparatus and heart. Cardiovascular malformations (CVMs) affecting the outflow tract (OFT) are frequently observed in 22q11.2DS and are among the most commonly occurring heart defects. The gene encoding T-box transcription factor 1 (Tbx1) has been identified as a major candidate for 22q11.2DS. However, CVMs are generally considered to have a multigenic basis and single-gene mutations underlying these malformations are rare. The T-box family members Tbx2 and Tbx3 are individually required in regulating aspects of OFT and pharyngeal development. Here, using expression and three-dimensional reconstruction analysis, we show that Tbx1 and Tbx2/Tbx3 are largely uniquely expressed but overlap in the caudal pharyngeal mesoderm during OFT development, suggesting potential combinatorial requirements. Cross-regulation between Tbx1 and Tbx2/Tbx3 was analyzed using mouse genetics and revealed that Tbx1 deficiency affects Tbx2 and Tbx3 expression in neural crest-derived cells and pharyngeal mesoderm, whereas Tbx2 and Tbx3 function redundantly upstream of Tbx1 and Hh ligand expression in pharyngeal endoderm and bone morphogenetic protein- and fibroblast growth factor-signaling in cardiac progenitors. Moreover, in vivo, we show that loss of two of the three genes results in severe pharyngeal hypoplasia and heart tube extension defects. These findings reveal an indispensable T-box gene network governing pharyngeal and OFT development and identify TBX2 and TBX3 as potential modifier genes of the cardiopharyngeal phenotypes found in TBX1-haploinsufficient 22q11.2DS patients.


Analysis of Response Elements Involved in the Regulation of the Human Neonatal Fc Receptor Gene (FCGRT).
Human epithelial, endothelial and PMA-differentiated THP-1 cell lines were used as model systems to study the transcriptional regulation of the human FCGRT gene encoding the alpha chain of hFcRn. The data obtained from site-directed mutagenesis in transient transfection experiments indicate that the Sp1 sites at positions -641, -635, and -313, CF1/YY1 elements at positions -586 and -357, and the AP-1 motif at -276 within the-660/-233 fragment of the human FCGRT promoter (hFCGRT) participate in the regulation of human FCGRT in all selected cell lines. However, their individual contribution to promoter activity is not equivalent. The Sp1 binding site at -313 and the AP-1 site at -276 are critical for the activity of the hFCGRT promoter in epithelial and endothelial cells. Moreover, the CF1/YY1 site at -586 in differentiated THP-1 cells, plays an essential role in the transcriptional activity of the promoter. In addition, the C/EBPbeta binding site at -497 of the hFCGRT promoter in epithelial and endothelial cells, and the C/EBPbeta motif located at -497 and -233 within the hFCGRT promoter in differentiated THP-1 cells may function as positive regulatory sequences in response to LPS or PMA stimulation. EMSA and supershift analyses showed that the functionally identified binding motifs in the hFCGRT promoter were able to specifically interact with their corresponding (Sp1, Sp2, Sp3, c-Fos, c-Jun, YY1, and C/EBPbeta or C/EBPdelta) transcription factors (TFs), suggesting their possible involvement in the regulation of the human FCGRT gene expression.


International Tuberculum Sellae Meningioma Study: Surgical Outcomes and Management Trends.
Tuberculum sellae meningiomas (TSMs) can be resected through transcranial (TCA) or expanded endonasal approach (EEA). The objective of this study was to report TSM management trends and outcomes in a large multicenter cohort. This is a 40-site retrospective study using standard statistical methods. In 947 cases, TCA was used 66.4% and EEA 33.6%. The median maximum diameter was 2.5 cm for TCA and 2.1 cm for EEA ( P < .0001). The median follow-up was 26 months. Gross total resection (GTR) was achieved in 70.2% and did not differ between EEA and TCA ( P = .5395). Vision was the same or better in 87.5%. Vision improved in 73.0% of EEA patients with preoperative visual deficits compared with 57.1% of TCA patients ( P < .0001). On multivariate analysis, a TCA (odds ratio [OR] 1.78, P = .0258) was associated with vision worsening, while GTR was protective (OR 0.37, P < .0001). GTR decreased with increased diameter (OR: 0.80 per cm, P = .0036) and preoperative visual deficits (OR 0.56, P = .0075). Mortality was 0.5%. Complications occurred in 23.9%. New unilateral or bilateral blindness occurred in 3.3% and 0.4%, respectively. The cerebrospinal fluid leak rate was 17.3% for EEA and 2.2% for TCA (OR 9.1, P < .0001). The recurrence rate was 10.9% (n = 103). Longer follow-up (OR 1.01 per month, P < .0001), World Health Organization II/III (OR 2.20, P = .0262), and GTR (OR: 0.33, P < .0001) were associated with recurrence. The recurrence rate after GTR was lower after EEA compared with TCA (OR 0.33, P = .0027). EEA for appropriately selected TSM may lead to better visual outcomes and decreased recurrence rates after GTR, but cerebrospinal fluid leak rates are high, and longer follow-up is needed. Tumors were smaller in the EEA group, and follow-up was shorter, reflecting selection, and observation bias. Nevertheless, EEA may be superior to TCA for appropriately selected TSM.


Molecular analysis of patterning of conduction tissues in the developing human heart.
Recent studies in experimental animals have revealed some molecular mechanisms underlying the differentiation of the myocardium making up the conduction system. To date, lack of gene expression data for the developing human conduction system has precluded valid extrapolations from experimental studies to the human situation. We performed immunohistochemical analyses of the expression of key transcription factors, such as ISL1, TBX3, TBX18, and NKX2-5, ion channel HCN4, and connexins in the human embryonic heart. We supplemented our molecular analyses with 3-dimensional reconstructions of myocardial TBX3 expression. TBX3 is expressed in the developing conduction system and in the right venous valve, atrioventricular ring bundles, and retro-aortic nodal region. TBX3-positive myocardium, with exception of the top of the ventricular septum, is devoid of fast-conducting connexin40 and connexin43 and hence identifies slowly conducting pathways. In the early embryonic heart, we found wide expression of the pacemaker channel HCN4 at the venous pole, including the atrial chambers. HCN4 expression becomes confined during later developmental stages to the components of the conduction system. Patterns of expression of transcription factors, known from experimental studies to regulate the development of the sinus node and atrioventricular conduction system, are similar in the human and mouse developing hearts. Our findings point to the comparability of mechanisms governing the development of the cardiac conduction patterning in human and mouse, which provide a molecular basis for understanding the functioning of the human developing heart before formation of a discrete conduction system.


Enhanced light trapping and high charge transmission capacities of novel structures for efficient photoelectrochemical water splitting.
Excellent PEC efficiency, good reusability and the super stability of trap-like SnS2/TiO2 nanotube arrays (NTs)-based photoanodes are reported. Specifically, the SnS2/TiO2-180 °C (ST-180) photoanode exhibited the highest photocurrent density (1.05 mA cm-2) and an optimal η (0.73%) at 0.5 V (vs. SCE) under simulated light irradiation (AM 1.5G), which are 4.6 and 3.8 times higher than those of pure TiO2 NTs (0.23 mA cm-2 and 0.19%). The IPCE values of ST-180 can reach 21.5% (365 nm) and 13.8% (420 nm), which are much higher than those of pure TiO2 NTs (10.6% at 365 nm and 0.8% at 420 nm). The APCE values of the pure TiO2 NTs photoelectrode are 12.8% (365 nm) and 1.1% (420 nm), while the ST-180 values are 22.3% and 14.2%, respectively. Furthermore, the generation rates of H2 and O2 for the ST-180 photoanode are 47.2 and 23.1 μmol cm-2 h-1 at 0.5 V under AM 1.5G, corresponding to faradaic efficiencies of around 80.1% and 78.3%, respectively. In short, the high-efficiency PEC water splitting performance of this SnS2/TiO2 photoanode results from the enhanced light harvesting ability of the trap-like SnS2 structure, accelerated carrier transportation properties of TiO2 NTs, and effective carrier separation of the type-II heterojunction structure. This work may offer a combinatorial strategy for the preparation of heterojunction structures with high PEC performance and can be a model structure for similar photoanode materials.


Bloating is associated with worse quality of life, treatment satisfaction, and treatment responsiveness among patients with constipation-predominant irritable bowel syndrome and functional constipation.
The management of bloating is unclear and its relationship with patients' well-being and treatment satisfaction independent of other abdominal symptoms is uncharacterized. We evaluated the association of bloating with patient-reported outcomes. Thirty-nine centers for functional gastrointestinal disorders joined the laxative inadequate relief survey. We enrolled 2203 consecutive outpatients with functional constipation (FC) or constipation-predominant irritable bowel syndrome (IBS-C) in two cross-sectional waves. Both wave 1 and 2 included the SF-12, the patient assessment of constipation-symptoms (PAC-SYM), and the treatment satisfaction questionnaire for medication (TSQM-2). Wave 2 only included a global rating of change (GRC) scale to assess patients' assessment of efficacy concerning treatment switches occurred in the 3 months prior to the interview. Bloating in the abdomen was defined on the basis of PAC-SYM item 3. The average age was 50.1 years (SD, 16.7) and 82.1% of patients were women. The prevalence of bloating was 91.6% (n = 1970). Bloating was associated with SF-12 Physical Composite Score (p < 0.01), SF-12 Mental Composite Score (p < 0.01), GRC (p < 0.01), Satisfaction with treatment effectiveness (p < 0.01), convenience of administration (p < 0.01), and side effects (p < 0.01) after adjustment for possible confounders. Our data suggest that patients regard bloating as a key element in assessing clinical changes and treatments' efficacy as this symptom exerts a strong influence on patient-reported outcomes independent of possible confounders and other symptoms of constipation. Our data provide the rationale to investigate the efficacy and tolerability of new treatments specifically addressing this important, yet disregarded, patients' complain.


Recent advances in drug repurposing using machine learning.
Drug repurposing aims to find new uses for already existing and approved drugs. We now provide a brief overview of recent developments in drug repurposing using machine learning alongside other computational approaches for comparison. We also highlight several applications for cancer using kinase inhibitors, Alzheimer's disease as well as COVID-19.


Atrial Anti-Arrhythmic Effects of Heptanol in Langendorff-Perfused Mouse Hearts.
Acute effects of heptanol (0.1 to 2 mM) on atrial electrophysiology were explored in Langendorff-perfused mouse hearts. Left atrial bipolar electrogram or monophasic action potential recordings were obtained during right atrial stimulation. Regular pacing at 8 Hz elicited atrial activity in 11 out of 11 hearts without inducing atrial arrhythmias. Programmed electrical stimulation using a S1S2 protocol provoked atrial tachy-arrhythmias in 9 of 17 hearts. In the initially arrhythmic group, 2 mM heptanol exerted anti-arrhythmic effects (Fisher's exact test, P < 0.05) and increased atrial effective refractory period (ERP) from 26.0 ± 1.9 to 57.1 ± 2.5 ms (ANOVA, P < 0.001) despite increasing activation latency from 18.7 ± 1.1 to 28.9 ± 2.1 ms (P < 0.001) and leaving action potential duration at 90% repolarization (APD90) unaltered (25.6 ± 1.2 vs. 27.2 ± 1.2 ms; P > 0.05), which led to increases in ERP/latency ratio from 1.4 ± 0.1 to 2.1 ± 0.2 and ERP/APD90 ratio from 1.0 ± 0.1 to 2.1 ± 0.2 (P < 0.001). In contrast, in the initially non-arrhythmic group, heptanol did not alter arrhythmogenicity, increased AERP from 47.3 ± 5.3 to 54.5 ± 3.1 ms (P < 0.05) and activation latency from 23.7 ± 2.2 to 31.3 ± 2.5 ms and did not alter APD90 (24.1 ± 1.2 vs. 25.0 ± 2.3 ms; P > 0.05), leaving both AERP/latency ratio (2.1 ± 0.3 vs. 1.9 ± 0.2; P > 0.05) and ERP/APD90 ratio (2.0 ± 0.2 vs. 2.1 ± 0.1; P > 0.05) unaltered. Lower heptanol concentrations (0.1, 0.5 and 1 mM) did not alter arrhythmogenicity or the above parameters. The present findings contrast with known ventricular pro-arrhythmic effects of heptanol associated with decreased ERP/latency ratio, despite increased ERP/APD ratio observed in both the atria and ventricles.


Encapsulating fly ash and acidic process waste water in brick structure.
Fly ash contains metals such as cadmium, iron, lead, aluminum and zinc in its structure in appreciable amounts. These metals can leach out into surface and ground waters if fly ash is not properly disposed of. A similar problem also exists for acidic process waste waters discharged by numerous industries. The purpose of this study was to utilize such wastes as additives in the production of construction quality bricks for the purpose of waste elimination. The bricks produced were subjected to flexural strength and water retention capacity tests along with heavy metal leaching experiments in order to determine the applicability of the procedure and the best possible recipes. This paper summarizes the results obtained in these tests along with the possible mechanisms involved in stabilizing the two wastes in the brick structure.


Heat shock factor 1 (HSF1) controls chemoresistance and autophagy through transcriptional regulation of autophagy-related protein 7 (ATG7).
Heat shock factor 1 (HSF1), a master regulator of heat shock responses, plays an important role in tumorigenesis. In this study we demonstrated that HSF1 is required for chemotherapeutic agent-induced cytoprotective autophagy through transcriptional up-regulation of autophagy-related gene ATG7. Interestingly, this is independent of the HSF1 heat shock response function. Treatment of cancer cells with the FDA-approved chemotherapeutic agent carboplatin induced autophagy and growth inhibition, which were significantly increased upon knockdown of HSF1. Mechanistic studies revealed that HSF1 regulates autophagy by directly binding to ATG7 promoter and transcriptionally up-regulating its expression. Significantly, breast cancer patient sample study revealed that a higher ATG7 expression level is associated with poor patient survival. This novel finding was further confirmed by analysis of two independent patient databases, demonstrating a prognostic value of ATG7. Furthermore, a strong positive correlation was observed between levels of HSF1 and ATG7 in triple-negative breast cancer patient samples, thus validating our in vitro findings. This is the first study identifying a critical role for HSF1 in controlling cytoprotective autophagy through regulation of ATG7, which is distinct from the HSF1 function in the heat shock response. This is also the first study demonstrating a prognostic value of ATG7 in breast cancer patients. These findings strongly argue that combining chemotherapeutic agents with autophagy inhibition by repressing HSF1/ATG7 axis represents a promising strategy for future cancer treatment.


Evolutionary dissection of mtDNA hg H: a susceptibility factor for hypertrophic cardiomyopathy.
Mitochondrial DNA (mtDNA) haplogroup (hg) H has been reported as a susceptibility factor for hypertrophic cardiomyopathy (HCM). This was established in genetic association studies, however, the SNP or SNP's that are associated with the increased risk have not been identified. Hg H is the most frequent European mtDNA hg with greater than 80 subhaplogroups (subhgs) each defined by specific SNPs. We tested the hypothesis that the distribution of H subhgs might differ between HCM patients and controls. The subhg H distribution in 55 HCM index cases was compared to that of two Danish mtDNA hg H control groups (n = 170 and n = 908, respectively). In the HCM group, H and 12 different H subhgs were found. All these, except subhgs H73, were also found in both control groups. The HCM group was also characterized by a higher proportion of H3 compared to H2. In the HCM group the H3/H2 proportion was 1.7, whereas it was 0.45 and 0.54 in the control groups. This tendency was replicated in an independent group of Hg H HCM index cases (n = 39) from Queensland, Australia, where the H3/H2 ratio was 1.5. In conclusion, the H subhgs distribution differs between HCM cases and controls, but the difference is subtle, and the understanding of the pathogenic significance is hampered by the lack of functional studies on the subhgs of H.


An outcome measure for patients with cervical myelopathy: Japanese Orthopaedic Association Cervical Myelopathy Evaluation Questionnaire (JOACMEQ): Part 1.
An outcome measure to evaluate the neurological function of cervical myelopathy was proposed by the Japanese Orthopaedic Association in 1975 (JOA score), and has been widely used in Japan. However, the JOA score does not include patients' satisfaction, disability, handicaps, or general health, which can be affected by cervical myelopathy. The purpose of this study was to develop a new outcome measure for patients with cervical myelopathy. This study was conducted in eight university hospitals and their affiliated hospitals from February to May 2002. The questionnaire included 77 items. Forty-one questions, which were originally listed by the authors, were for evaluation of the physical function of the cervical spine and spinal cord. The Medical Outcome Study Short-Form 36-Item Health Survey (SF-36) was used to examine health-related quality of life (QOL). Patients with cervical myelopathy and healthy volunteers were recruited at each institution. After analysis of the answers from patients and volunteers, irrelevant questions using the following criteria were excluded: (1) a question 80% of answers for which were concentrated on one choice, (2) a question whose answer was highly correlated with that of other questions, (3) a question that could be explained by other questions, and (4) a question for which the distribution of the answers obtained from the patients was not different from that obtained from the normal volunteers. The patients comprised 164 men and 86 women, and the healthy volunteers 96 men and 120 women. Thirteen items from the questions about the physical functions of the cervical spine and the spinal cord and 11 items from SF-36 remained as candidates that should be included in the final outcome measure questionnaire. Twenty-four questions remained as candidates for the final questionnaire. This new self-administered questionnaire might be used to evaluate the outcomes in patients with cervical myelopathy more efficiently.


Surface tension propulsion of fungal spores.
Most basidiomycete fungi actively eject their spores. The process begins with the condensation of a water droplet at the base of the spore. The fusion of the droplet onto the spore creates a momentum that propels the spore forward. The use of surface tension for spore ejection offers a new paradigm to perform work at small length scales. However, this mechanism of force generation remains poorly understood. To elucidate how fungal spores make effective use of surface tension, we performed a detailed mechanical analysis of the three stages of spore ejection: the transfer of energy from the drop to the spore, the work of fracture required to release the spore from its supporting structure and the kinetic energy of the spore after ejection. High-speed video imaging of spore ejection in Auricularia auricula and Sporobolomyces yeasts revealed that drop coalescence takes place over a short distance ( approximately 5 microm) and energy transfer is completed in less than 4 mus. Based on these observations, we developed an explicit relation for the conversion of surface energy into kinetic energy during the coalescence process. The relation was validated with a simple artificial system and shown to predict the initial spore velocity accurately (predicted velocity: 1.2 m s(-1); observed velocity: 0.8 m s(-1) for A. auricula). Using calibrated microcantilevers, we also demonstrate that the work required to detach the spore from the supporting sterigma represents only a small fraction of the total energy available for spore ejection. Finally, our observations of this unique discharge mechanism reveal a surprising similarity with the mechanics of jumping in animals.


Genetic diversity in clinical isolates of the dimorphic fungus Blastomyces dermatitidis detected by a PCR-based random amplified polymorphic DNA assay.
Blastomyces dermatitidis is a dimorphic fungus causing localized or systemic infection in areas where the organism is endemic in the central and southeastern United States. In this study, 19 independent isolates of B. dermatitidis from Little Rock, Ark., were grouped into three classes based on restriction fragment length polymorphism patterns in mitochondrial DNA with a heterologous probe from Histoplasma capsulatum. One large class of 15 isolates and two smaller classes (classes 2 and 3), each consisting of two isolates, were observed in BglII digests. Strain-specific arrays of PCR-amplified DNA products were obtained with arbitrarily selected primers (18 to 29 nucleotides long; G+C contents, 33 to 56%). In the large class 1 group, 13 isolates could be differentiated by the random amplified polymorphic DNA (RAPD) method with various primers. The two remaining class 1 isolates were obtained from the same patients and produced identical RAPD arrays. Dissimilar RAPD patterns were obtained from the smaller class 2 group but not from the class 3 isolates. Significant genetic diversity in clinical isolates of B. dermatitidis was observed; this may underscore a similar environmental diversification. Further application of the typing techniques may provide significant insight into the epidemiology of blastomycosis and aid in the assessment of specific virulence phenotypes.


Dispensing errors and counseling quality in 100 pharmacies.
To evaluate the dispensing accuracy and counseling provided in community chain pharmacies. Cross-sectional study. Community chain pharmacies in large metropolitan areas of Florida, Georgia, New Jersey, and New York. Community chain pharmacies and trained shoppers. Trained shoppers presented a new prescription order for one of five study drugs to each randomly selected pharmacy, and all encounters with pharmacy staff were recorded on video by ABC News 20/20 staff using hidden cameras. Dispensing errors on prescriptions for selected medications were the indicator of prescription dispensing accuracy. Frequency of verbal counseling and information categories discussed or included in written information were used to assess the quality of counseling. Of 100 prescriptions dispensed, 22 had one or more deviation from the physician's written order, for a 22% dispensing error rate. Three of the errors were judged to be potentially harmful when dispensed to a typical patient requiring these therapies. A total of 43 shoppers (43%) received verbal counseling, including 16 cases in which the shopper prompted counseling. All shoppers received written information with their prescription, covering an average of 90% of the required topics. Some 68% of the warfarin shoppers purchased aspirin without the pharmacist verbally warning about taking the drugs simultaneously. The dispensing error rate of more than one in five prescriptions is similar to the rate found in a similar study conducted 14 years ago, but counseling frequency has decreased significantly during the period.


Peripheral nerve and neuromuscular junction pathology in Pompe disease.
Pompe disease is a systemic metabolic disorder characterized by lack of acid-alpha glucosidase (GAA) resulting in ubiquitous lysosomal glycogen accumulation. Respiratory and ambulatory dysfunction are prominent features in patients with Pompe yet the mechanism defining the development of muscle weakness is currently unclear. Transgenic animal models of Pompe disease mirroring the patient phenotype have been invaluable in mechanistic and therapeutic study. Here, we demonstrate significant pathological alterations at neuromuscular junctions (NMJs) of the diaphragm and tibialis anterior muscle as prominent features of disease pathology in Gaa knockout mice. Postsynaptic defects including increased motor endplate area and fragmentation were readily observed in Gaa(-/-) but not wild-type mice. Presynaptic neuropathic changes were also evident, as demonstrated by significant reduction in the levels of neurofilament proteins, and alterations in axonal fiber diameter and myelin thickness within the sciatic and phrenic nerves. Our data suggest the loss of NMJ integrity is a primary contributor to the decline in respiratory and ambulatory function in Pompe and arises from both pre- and postsynaptic pathology. These observations highlight the importance of systemic phenotype correction, specifically restoration of GAA to skeletal muscle and the nervous system for treatment of Pompe disease.


Macrophages enhance mesenchymal stem cell osteogenesis via down-regulation of reactive oxygen species.
The role played by macrophages in regulating the differentiation of mesenchymal stem cells (MSCs) during wound healing and bone regeneration is increasingly being recognized. The present study compared the pro-osteogenic effects of three co-culture methods, conditioned medium generated by macrophages (CM), indirect culture (IC) or direct culture (DC) with macrophages, on bone marrow MSCs (BMMSCs). Primary BMMSCs were isolated, characterized and co-cultured with RAW264.7 mouse macrophages. Cell morphology and intracellular reactive oxygen species (ROS) levels were determined by scanning electron microscopy (SEM) and flow cytometry, respectively. Alkaline phosphatase (ALP) staining and assay, Alizarin red staining (ARS) and quantitative real-time polymerase chain reaction (qRT-PCR) were performed to evaluate osteogenic differentiation. Inclusion of macrophages in any of the three co-culture methods resulted in improvement in osteogenic differentiation and mineralization of BMMSCs (DC > IC > CM), as measured by ALP staining and activity, ARS and osteoblastic gene expression (Runx2, Alp, Ocn and Bmp2). The enhanced osteogenesis was reversed with hydrogen peroxide. Macrophages reduced the increased levels of intracellular ROS generated by BMMSCs during osteogenic differentiation in a manner similar to the use of an antioxidant, N-acetyl cysteine. Macrophages exert an osteogenesis-enhancing effect to accelerate BMMSC osteogenesis via ROS downregulation. The present findings suggest that targeting MSC-macrophage interaction is an effective strategy for regulating stem cell fate and facilitating bone regeneration.


Molecular Mechanisms Underlying the Pain-Relieving Effects of Extracorporeal Shock Wave Therapy: A Focus on Fascia Nociceptors.
In recent years, extracorporeal shock wave therapy (ESWT) has received increasing attention for its potential beneficial effects on various bone and soft-tissue pathologies, yielding promising outcomes for pain relief and functional recovery. In fact, ESWT has emerged as an alternative, non-invasive, and safe treatment for the management of numerous musculoskeletal disorders, including myofascial pain syndrome (MPS). In particular, MPS is a common chronic painful condition, accounting for the largest proportion of patients affected by musculoskeletal problems. Remarkably, sensory innervation and nociceptors of the fascial system are emerging to play a pivotal role as pain generators in MPS. At the same time, increasing evidence demonstrates that application of ESWT results in selective loss of sensory unmyelinated nerve fibers, thereby inducing long-lasting analgesia. The findings discussed in the present review are supposed to add novel viewpoints that may further enrich our knowledge on the complex interactions occurring between disorders of the deep fascia including changes in innervation, sensitization of fascial nociceptors, the pathophysiology of chronic musculoskeletal pain of MPS, and EWST-induced analgesia. Moreover, gaining mechanistic insights into the molecular mechanisms of pain-alleviating effects of ESWT may broaden the fields of shock waves clinical practice far beyond the musculoskeletal system or its original application for lithotripsy.


p53 mediates apoptosis induced by c-Myc activation in hypoxic or gamma irradiated fibroblasts.
Deregulated c-Myc expression leads to a cellular state where proliferation and apoptosis are equally favored depending on the cellular microenvironment. Since the apoptotic sensitivity of many cells is influenced by the status of the p53 tumor suppressor gene, we investigated whether the induction of apoptosis by DNA damage or non-genotoxic stress are also influenced by the p53 status of cells with altered c-Myc activity. Rat-1 fibroblasts expressing a conditional c-Myc allele (c-MycER), were transfected to express an antisense RNA complimentary to p53 mRNA. Expression of antisense p53 RNA decreased p53 protein levels and delayed p53 accumulation following c-Myc activation. Under hypoxic or low serum conditions, cells expressing antisense p53 were substantially more resistant to c-Myc-induced apoptosis than were control cells. c-Myc activation also sensitized Rat-1 cells to radiation-induced apoptosis. Rat-1 cells expressing antisense p53 RNA were more resistant to apoptosis induced by the combined effects of c-Myc activation and gamma irradiation. In a similar manner, apoptosis induced by c-Myc in serum starved, hypoxic or gamma irradiated fibroblasts was also inhibited by Bcl-2. These data indicate that p53 is involved in c-Myc-mediated apoptosis under a variety of stresses which may influence tumor growth, evolution and response to therapy.


[The influence of acidic conditions on polar exopolysaccharides which bind Rhizobium japonicum to soybean root hairs].
The influence of acidic conditions on rhizobial exopolysaccharides (EPS) and their symbiotic association in host plants, were investigated. Light micrographs of soybean root hairs were taken with a Fahraeus' slide assembly after the inocubation of rhizobial bacteria; curled and swollen root hairs, and the typical shepherd's crook with an infection thread were observed. The colonizations of soybean root hairs by rhizobia were different under acidic and neutral conditions. For the Rhizobium japonicum strain USDA 136, and the indigenous acidic soil strain KR 23, morphologically distinct types of cells occurred in the acidic and neutral yeast-extract-mannitol broths. The surface structure of R. japonicum, as examined with the carbon replica and negative stain techniques, was rod shaped and polar flagellated only when the cells were incubated under neutral conditions. The materials which reacted with the ruthenium red positive reagent are in all likelihood an acidic EPS, and are believed to effect the adhesive properties of the cells. Bacterial polarity, which could be more readily observed under neutral conditions than under acidic conditions, was related to the distribution of EPS near one end of the bacteria. EPS was found to effect the initial step of polar attachment in the effective infection of host roots. By using DEAE-Sephacel ion exchange chromatography, the capsular polysaccharides from both strains of R. japonicum were also found to have different chemical components when observed in acidic and neutral cultures.


Ribosomal profiling during prion disease uncovers progressive translational derangement in glia but not in neurons.
Prion diseases are caused by PrPSc, a self-replicating pathologically misfolded protein that exerts toxicity predominantly in the brain. The administration of PrPSc causes a robust, reproducible and specific disease manifestation. Here, we have applied a combination of translating ribosome affinity purification and ribosome profiling to identify biologically relevant prion-induced changes during disease progression in a cell-type-specific and genome-wide manner. Terminally diseased mice with severe neurological symptoms showed extensive alterations in astrocytes and microglia. Surprisingly, we detected only minor changes in the translational profiles of neurons. Prion-induced alterations in glia overlapped with those identified in other neurodegenerative diseases, suggesting that similar events occur in a broad spectrum of pathologies. Our results suggest that aberrant translation within glia may suffice to cause severe neurological symptoms and may even be the primary driver of prion disease.


Contribution of SARS-CoV-2 Accessory Proteins to Viral Pathogenicity in K18 Human ACE2 Transgenic Mice.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the viral pathogen responsible for the current coronavirus disease 2019 (COVID-19) pandemic. As of 19 May 2021, John Hopkins University's COVID-19 tracking platform reported 3.3 million deaths associated with SARS-CoV-2 infection. Currently, the World Health Organization has granted emergency use listing (EUL) to six COVID-19 vaccine candidates. However, much of the pathogenesis observed during SARS-CoV-2 infection remains elusive. To gain insight into the contribution of individual accessory open reading frame (ORF) proteins in SARS-CoV-2 pathogenesis, we used our recently described reverse-genetics system approach to successfully engineer recombinant SARS-CoV-2 (rSARS-CoV-2) constructs; we removed individual viral ORF3a, -6, -7a, -7b, and -8 proteins from them, and we characterized the resulting recombinant viruses in vitro and in vivo. Our results indicate differences in plaque morphology, with ORF-deficient (ΔORF) viruses producing smaller plaques than those of the wild type (rSARS-CoV-2/WT). However, growth kinetics of ΔORF viruses were like those of rSARS-CoV-2/WT. Interestingly, infection of K18 human angiotensin-converting enzyme 2 (hACE2) transgenic mice with the ΔORF rSARS-CoV-2s identified ORF3a and ORF6 as the major contributors of viral pathogenesis, while ΔORF7a, ΔORF7b, and ΔORF8 rSARS-CoV-2s induced pathology comparable to that of rSARS-CoV-2/WT. This study demonstrates the robustness of our reverse-genetics system to generate rSARS-CoV-2 constructs and the major role for ORF3a and ORF6 in viral pathogenesis, providing important information for the generation of attenuated forms of SARS-CoV-2 for their implementation as live attenuated vaccines for the treatment of SARS-CoV-2 infection and associated COVID-19. IMPORTANCE Despite great efforts put forward worldwide to combat the current coronavirus disease 2019 (COVID-19) pandemic, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to be a human health and socioeconomic threat. Insights into the pathogenesis of SARS-CoV-2 and the contribution of viral proteins to disease outcome remain elusive. Our study aims (i) to determine the contribution of SARS-CoV-2 accessory open reading frame (ORF) proteins to viral pathogenesis and disease outcome and (ii) to develop a synergistic platform combining our robust reverse-genetics system to generate recombinant SARS-CoV-2 constructs with a validated rodent model of infection and disease. We demonstrate that SARS-CoV-2 ORF3a and ORF6 contribute to lung pathology and ultimately disease outcome in K18 hACE2 transgenic mice, while ORF7a, ORF7b, and ORF8 have little impact on disease outcome. Moreover, our combinatory platform serves as a foundation for generating attenuated forms of the virus to develop live attenuated vaccines for the treatment of SARS-CoV-2.


Single-molecule force spectroscopy using the NanoTracker optical tweezers platform: from design to application.
Since the development of detection and analysis techniques for optical tweezers setups, there has been an ever-increasing interest in optical tweezers as a quantitative method, shifting its applications from a pure manipulation tool towards the investigation of motions and forces. With the capability of manipulation and detection of forces of a few hundred picoNewtons down to a fraction of a picoNewton, optical tweezers are perfectly suitable for the investigation of single molecules. Accordingly, the technique has been extensively used for the biophysical characterization of biomolecules, ranging from the mechanical and elastic properties of biological polymers to the dynamics associated with enzymatic activity and protein motility. Here, the use of state-of-the-art optical tweezers on the elasticity of single DNA molecules is presented, highlighting the possibilities this technique offers for the investigation of protein-DNA interaction, but also for other single molecule applications. Technical in nature, design aspects of the NanoTracker optical tweezers setup are addressed, presenting the recent advances in the development of optical tweezers, ranging from noise reduction to detection and calibration methodology.


Persistent repression of a functional allele can be responsible for galactosyltransferase deficiency in Tn syndrome.
A human hematopoietic disorder designated as Tn syndrome or permanent mixed-field polyagglutinability has been ascribed to a stem cell mutation leading to a specific deficiency of UDP-Gal:GalNAc alpha 1-O-Ser/Thr beta 1-3 galactosyltransferase (beta 3 Gal-T) activity in affected cells. To test for the possibility that an allele of the beta 3Gal-T gene might be repressed instead of mutated, we have investigated whether 5-azacytidine or sodium n-butyrate, both inducers of gene expression, would reactivate expression of beta 3Gal-T in cloned enzyme-deficient T cells derived from a patient affected by the Tn syndrome. Flow cytometry revealed that a single treatment induced de novo expression of the Thomsen-Friedenreich antigen (Gal beta 1-3GalNAc-R), the product of beta 3Gal-T activity. In addition, a sialylated epitope on CD43 (leukosialin), which is present on normal but not on beta 3Gal-T-deficient T cells, was also reexpressed. Although no beta 3Gal-T activity was detectable in untreated Tn syndrome T cells, after exposure to 5-azaC,beta 3Gal-T activity reached nearly normal values. Both agents failed to reactivate beta 3Gal-T in Jurkat T leukemic cells, which also lack beta 3Gal-T activity. These data demonstrate that Tn syndrome T cells contain an intact beta 3Gal-T gene copy and that the enzyme deficiency in this patient is due to a persistent and complete but reversible repression of a functional allele. In contrast, the cause of beta 3Gal-T deficiency appears to be different in Jurkat T cells.


Aberrant B-cell activation and B-cell subpopulations in rheumatoid arthritis: analysis by clinical activity, autoantibody seropositivity, and treatment.
Few studies analyze the role of B-cell subpopulations in rheumatoid arthritis (RA) pathophysiology. Therefore, this study aimed to analyze the differences in B-cell subpopulations and B-cell activation according to disease activity, RA subtype, and absence of disease-modifying antirheumatic drugs (DMARDs) therapy. These subgroups were compared with control subjects (CS). One hundred and thirty-nine subjects were included, of which 114 were RA patients, and 25 were controls. Patients were divided into 99 with seropositive RA, 6 with seronegative RA, and 9 without DMARDs. The patients with seropositive RA were subclassified based on the DAS28 index. A seven-color multicolor flow cytometry panel was used to identify B-cell immunophenotypes and cell activation markers. There were no changes in total B-cell frequencies between RA patients and controls. However, a lower frequency of memory B cells and pre-plasmablasts was observed in seropositive RA compared to controls (P < 0.0001; P = 0.0043, respectively). In contrast, a higher frequency of mature B cells was observed in RA than in controls (P = 0.0002). Among patients with RA, those with moderate activity had a higher percentage of B cells (P = 0.0021). The CD69+ marker was increased (P < 0.0001) in RA compared to controls, while the CD40+ frequency was decreased in patients (P < 0.0001). Transitional, naïve, and double-negative B-cell subpopulations were higher in seronegative RA than in seropositive (P < 0.01). In conclusion, in seropositive and seronegative RA patients, there are alterations in B-cell activation and B-cell subpopulations, independently of clinical activity and DMARDs therapy.


Interface magnetism in epitaxial BiFeO3-La0.7Sr0.3MnO3 heterostructures integrated on Si(100).
We report on the heteroepitaxial growth of ferroelectric (FE)-antiferromagnetic (AFM) BiFeO3 (BFO) on ferromagnetic La0.7Sr0.3MnO3 (LSMO), integrated on Si(100) using pulsed laser deposition via the domain matching epitaxy paradigm. The BFO/LSMO films were epitaxially grown on Si(100) by introducing epitaxial layers of SrTiO3/MgO/TiN. X-ray diffraction, scanning electron microscopy, high-resolution transmission electron microscopy, X-ray photo absorption spectroscopy, and atomic force microscopy were employed to fully characterize the samples. Furthermore, we have investigated the magnetic behavior of this five layer heterostructure, in which a d(5) system (Fe(3+)) manifested in FE-AFM BFO is epitaxially conjoined at the interface to a multivalent transition metal ion such as Mn(3+)/Mn(4+) in LSMO. The temperature- and magnetic field-dependent magnetization measurements reveal an unexpected enhancement in magnetic moment and improved magnetic hysteresis squareness originating from the BFO/LSMO interface. We observe a stronger temperature dependence of HEB when the polarity of field cooling is negative as compared to positive field cooling. We believe such an enhancement in magnetic moment and magnetic coupling is likely directly related to an electronic orbital reconstruction at the interface and complex interplay between orbital and spin degrees of freedom, similar to what has previously been reported in the literature. Future work will involve the linearly polarized X-ray absorption measurements to prove this hypothesis. This work represents a starting step toward the realization of magneto-electronic devices integrated with Si(100).


Blindness to a simultaneous change of all elements in a scene, unless there is a change in summary statistics.
Sudden change of every object in a display is typically conspicuous. We find however that in the presence of a secondary task, with a display of moving dots, it can be difficult to detect a sudden change in color of all the dots. A field of 200 dots, half red and half green, half moving rightward and half moving leftward, gave the appearance of two surfaces. When all 200 dots simultaneously switched color between red and green, performance in detecting the switch was very poor. A key display characteristic was that the color proportions on each surface (summary statistics) were not affected by the color switch. When the color switch is accompanied by a change in these summary statistics, people perform well in detecting the switch, suggesting that the secondary task does not disrupt the availability of this statistical information. These findings suggest that when the change is missed, the old and new colors were represented, but the color-location pattern (binding of colors to locations) was not represented or not compared. Even after extended viewing, changes to the individual color-location pattern are not available, suggesting that the feeling of seeing these details is misleading.


Monitoring paired binary surgical outcomes using cumulative sum charts.
Correlated binary data are encountered in many areas of medical research, system reliability and quality control. For monitoring failures rates in such situations, simultaneous bivariate cumulative sum (CUSUM) charts with the addition of secondary control limits are proposed. Using an approach based on a Markov chain model, the run length properties of such a monitoring scheme can be determined for sudden, or gradual, changes in the failure rates. The proposed control charts are easy to implement, and are shown to be very effective at detecting small changes in the rate of undesirable outcomes, especially when the changes are gradual. This procedure is illustrated using bivariate outcome data arising from a series of paediatric surgeries. The methodology is sufficiently general that it may be adapted for multivariate normal, binomial or Poisson responses.


Genomic organization, promoter activity, and expression of the human choline transporter-like protein 1.
Choline transporter-like (CTL) proteins of the CTL1 family are novel transmembrane proteins implicated in choline transport for phospholipid synthesis. In this study, we characterized the 5'-flanking region of the human (h)CTL1 gene and examined some of the possible mechanisms of its regulation, including promoter activity, splicing, and expression. The transcription start site of the hCTL1 gene was mapped by 5'-rapid amplification of cDNA ends (RACE), and the presence of two splice variants, hCTL1a and hCTL1b, was investigated using isoform-specific PCR and 3'-RACE. The hCTL1 promoter region of approximately 900 bp was isolated from MCF-7 human breast cancer cells. The promoter was TATA-less and driven by a long stretch of GC-rich sequence in accordance with widespread expression of hCTL1 at both mRNA and protein levels. Deletion analyses demonstrated that a very strong promoter is contained within 500 bp of the transcription start site, and more upstream regions did not increase its activity. The core promoter that conferred the minimal transcription is within the -188/+27-bp region, and its activity varied in human breast cancer and mouse skeletal muscle cells. Multiple motifs within the promoter regulatory region bound nuclear factors from both cultured cells and normal human skeletal muscle. The motifs within the three regions [S1 (-92/-61 bp), S2 (-174/-145 bp), and S3 (-289/-260 bp)] contained overlapping binding sites for hematopoietic transcription factors and ubiquitous transcription factors, in line with the expected gene function. Genomic analyses demonstrated a high conservation of hCTL1 and mouse CTL1 proximal promoters. Accordingly, mRNA profiles demonstrated that human splice variants were expressed ubiquitously, as demonstrated for the mouse transcripts; however, they differed from the profiles of rat CTL1 transcripts, which were more restricted to neurons and intestinal tissues. The shorter hCTL1b variant contained the cytosolic COOH-terminal motif L651KKR654 for endoplasmic reticulum retrieval/retention. This retention signal was conserved in hCTL1b and rat and mouse CTL1b and is typical for transmembrane proteins of type 1 topology.


Synthesis of Nitro-Aryl Functionalised 4-Amino-1,8-Naphthalimides and Their Evaluation as Fluorescent Hypoxia Sensors.
Fluorescent sensors are a vital research tool, enabling the study of intricate cellular processes in a sensitive manner. The design and synthesis of responsive and targeted probes is necessary to allow such processes to be interrogated in the cellular environment. This remains a challenge, and requires methods for functionalisation of fluorophores with multiple appendages for sensing and targeting groups. Methods to synthesise more structurally complex derivatives of fluorophores will expand their potential scope. Most known 4-amino-1,8-naphthalimides are only functionalised at imide and 4-positions, and structural modifications at additional positions will increase the breadth of their utility as responsive sensors. In this work, methods for the incorporation of a hypoxia sensing group to 4-amino-1,8-naphthalimide were evaluated. An intermediate was developed that allowed us to incorporate a sensing group, targeting group, and ICT donor to the naphthalimide core in a modular fashion. Synthetic strategies for attaching the hypoxia sensing group and how they affected the fluorescence of the naphthalimide were evaluated by photophysical characterisation and time-dependent density functional theory. An extracellular hypoxia probe was then rationally designed that could selectively image the hypoxic and necrotic region of tumour spheroids. Our results demonstrate the versatility of the naphthalimide scaffold and expand its utility. This approach to probe design will enable the flexible, efficient generation of selective, targeted fluorescent sensors for various biological purposes.


A preliminary report of a surveillance scheme of occupational asthma in the West Midlands.
The results from the first year of a notification scheme for occupational asthma in the West Midlands Region are presented. The scheme includes recognised new and old cases of occupational asthma. Thirty new cases were recognised per million general working population in the first year. Cases recognised in different occupational groups ranged from 154 per million painters and assembly workers to three per million clerical staff. Analysis of the agents to which workers with recognised occupational asthma were exposed identified commonly recognised agents such as isocyanates, colophony, and flour and generally less well recognised ones such as oil mists. The distribution of new and old reported cases, including those receiving compensation from the Department of Social Security, were calculated by health authority using estimated working population as the denominator. The number of reported cases varied from 303 per million in a semi-urban health authority that has a respiratory physician with a special interest in occupational asthma to less than 30 cases per million in eight health authorities. The most likely cause for these differences is lack of ascertainment.


[Hypodense eosinophils in blood of pediatric patients with asthma].
Determine the number of circulating hypodense eosinophils in the blood of atopic asthmatics. The hypothesis was that hypodense eosinophils, the activated cells, were higher in asthmatics than in non-atopic children. Granulocytes were separated from mononuclear cells by Ficoll-hypaque. Gradients of density were generated through dilutions of Percoll for fractionations of cells. Asthmatics had blood eosinophilia (768 +/- 75/mcl) when compared to normal control (200 +/- 28/mcl), and higher mean hypodense eosinophils (39%) as compared to normal children (11.7%). Eosinophils accumulate in asthmatic, especially the low density activated variant, suggesting its active role in the allergic inflammatory process.


Hepatic lesion characterization in cirrhosis: significance of arterial hypervascularity on dual-phase helical CT.
Our goal was to evaluate the diagnostic significance of the presence and pattern of arterial hypervascularity in lesions detected on dual-phase helical CT in cirrhotic patients. Fifty-eight lesions greater than 1 cm in size were prospectively identified in 26 patients with end-stage liver disease who had undergone dual-phase helical CT for preoperative liver transplantation evaluation. All 26 patients had diagnoses proven by histologic evaluation or by clinical criteria. All arterial phase scans were retrospectively reviewed and lesions were categorized for the presence and pattern of arterial hypervascularity. Radiologic findings were correlated with histopathologic data. Thirty-seven of the 58 lesions had hypervascular components on arterial phase scans. All 37 of these lesions were found to represent hepatocellular carcinoma (HCC) (positive predictive value, 100%). Of the 21 remaining hypovascular lesions, 17 were HCC and four were benign (positive predictive value, 81%). Of the nine patients in whom all lesions were hypovascular, six had HCC (positive predictive value, 66%). The value of the presence of arterial hypervascularity for diagnosing HCC was statistically significant (p < .05). However, the presence or absence of arterial hypervascularity and the specific enhancement pattern revealed by helical CT did not correlate with histologic grading. The presence of hypervascularity in hepatic masses found in cirrhotic patients is highly predictive of malignancy.


Quantifying the regional variations in the mechanical properties of cancellous bone of the tibia using indentation testing and quantitative computed tomographic imaging.
Finite element models apply material properties using experimentally derived density-modulus equations and computed tomographic image data, yet numerous different equations exist in the literature. The purpose of this study was to experimentally evaluate the distribution of mechanical properties through the proximal tibia and compare with those predicted using existing density-modulus equations. Indentation testing was performed on five cadaveric tibiae, with four slices removed from the proximal epiphysis and metaphysis of each. Elastic modulus and yield strength were identified for each test and grouped into nine transverse regions. These regions were identified on computed tomographic scans, and four density-modulus equations from the literature applied. Errors between measured and predicted modulus were then calculated. Elastic modulus and yield strength varied regionally, with the bone located closest to the joint and in the condyles being strongest and the intercondylar region the weakest. The optimal relationship for predicting modulus varied depending on anatomical region, but generally was best predicted by the Goulet equation. The regions of high strength identified in this study (condyles and proximal regions) can serve as improved sites of attachment for orthopedic devices and should be preserved during surgery, if possible. The substantial regional variations observed herein (almost a threefold change in modulus across different regions) should be incorporated into finite element models and applied using the Goulet density-modulus equation.


Impact of the COVID-19 pandemic on human salmonellosis in the Netherlands.
The public health measures implemented to control coronavirus disease 2019 (COVID-19) may influence also other infectious diseases. Using national laboratory surveillance data, we assessed the impact of the COVID-19 pandemic on human salmonellosis in the Netherlands until March 2021. Salmonellosis incidence decreased significantly after March 2020: in the second, third and fourth quarters of 2020, and in the first quarter of 2021, the incidence decreased by 55%, 57%, 47% and 37%, respectively, compared to the same quarters of 2016-2019. The decrease was strongest among travel-related cases (94%, 84%, 79% and 93% in the aforementioned quarters, respectively). Other significant changes were: increased proportion of cases among older adults and increased proportion of invasive infections, decreased proportion of trimethoprim resistance and increased proportion of serovar Typhimurium monophasic variant vs. Enteritidis. This led to decreased contributions of laying hens and increased contributions of pigs and cattle as sources of human infections. The observed changes probably reflect a combination of reduced exposure to Salmonella due to restrictions on international travels and gatherings, closure of dine-in restaurants, catering and hospitality sectors at large and changes in healthcare-seeking and diagnostic behaviours.


Towards personalized therapy for patients with glioblastoma.
Combined therapy with temozolomide and radiotherapy is a standard treatment and improves the survival for patients with newly diagnosed glioblastoma. However, the prognosis remains poor, with a median survival time of 12-15 months. Currently, several clinical trials of dose-dense temozolomide regimen or molecular-targeting therapies have been performed to overcome the resistance of glioblastoma. In these therapies, rational prognostic biomarkers have also been investigated to predict their outcome and response to treatment. This advanced understanding of the biological markers can help to develop personalized therapies for glioblastoma patients. Generally, due to a reduced tolerance, elderly patients do not seem to benefit from intensive treatment. This population needs individual treatments depended on their age or performance status. In this article, we review the recent studies that can provide personalized therapy for glioblastoma, based on molecular tumor profiling or patients' physical status.


Fluorescence microscopic studies of mitochondrial nucleoids during meiosis and sporulation in the yeast, Saccharomyces cerevisiae.
Configurational changes of mitochondria and mitochondrial nucleoids (mt-nucleoids) during meiosis and sporulation in the yeast, Saccharomyces cerevisiae, were examined using the mitochondrial membrane-binding fluorescent dye, dimethyl aminostyrylmethylpyridiniumiodine (DASPMI) and the DNA-binding fluorescent dye, 4',6-diamidino-2-phenylindole (DAPI). In zygotes just after mating, mt-nucleoids were observed as many small discrete light spots in the cytoplasm. During meiosis in zygotes, mt-nucleoids at first coalesced with each other into a long string and then separated into spherical nucleoids in four spores. These changes paralleled those in mitochondria observed using DASPMI. The use of spheroplasts allowed us to examine the behaviour of mt-nucleoids at higher resolution and to identify several distinct meiotic prophase stages of the cell nucleus during early sporulation. In diploid spheroplasts at the stationary phase, 50-70 of the mt-nucleoids were observed to be separated from each other and each spherical mitochondrion contained only one mt-nucleoid. At the later stage of premeiotic DNA synthesis, a single branched giant mitochondrion was formed as a result of complete mitochondrial fusion. All of the mt-nucleoids were arranged in an array on a giant mitochondrion and coalesced into a string-like network. Through meiosis I and II, strings of mt-nucleoids were observed close to the dividing nuclei. At late meiosis II, a ring of mt-nucleoids enclosing each daughter nucleus was formed. In ascospores, discrete small nucleoids were visible close to each spore nucleus with a 'string-of-beads' appearance. Many mt-nucleoids were excluded from the ascospores and remained in the residual cytoplasm of the ascus.


Single-band amplitude demodulation of Müller-Lyer illusion images.
The perception of the Müller-Lyer illusion has previously been explained as a result of visual low band-pass spatial filtering, although, in fact, the illusion persists in band-pass and high-pass filtered images without visible low-spatial frequencies. A new theoretical framework suggests that our perceptual experience about the global spatial structure of an image corresponds to the amplitude modulation (AM) component (or its magnitude, also called envelope) of its AM-FM (alternatively, AM-PM) decomposition. Because demodulation is an ill-posed problem with a non-unique solution, two different AM-FM demodulation algorithms were applied here to estimate the envelope of images of Müller-Lyer illusion: the global and exact Daugman and Downing (1995) AMPM algorithm and the local and quasi-invertible Maragos and Bovik (1995) DESA. The images used in our analysis include the classic configuration of illusion in a variety of spatial and spatial frequency content conditions. In all cases, including those of images for which visual low-pass spatial filtering would be ineffective, the envelope estimated by single-band amplitude demodulation has physical distortions in the direction of perceived illusion. It is not plausible that either algorithm could be implemented by the human visual system. It is shown that the proposed second order visual model of pre-attentive segregation of textures (or "back-pocket" model) could recover the image envelope and, thus, explain the perception of this illusion even in Müller-Lyer images lacking low spatial frequencies.


Efficacy and safety of tenofovir disoproxil fumarate in preventing vertical transmission of hepatitis B in pregnancies with high viral load.
This study was a meta-analysis of the literature on the efficacy and safety of tenofovir disoproxil fumarate (TDF) in preventing vertical transmission of hepatitis B in pregnancies with high viral load. Four observational studies and one randomized controlled trial involving 585 pregnant women and 595 newborns were included in the meta-analysis. TDF was more effective than the placebo in reducing vertical transmission in HBeAg-positive chronic hepatitis B (CHB) pregnancies with high serum HBV-DNA levels (OR = 0.21, 95% CI = 0.07-0.61) at 4-12 months, infant HBV DNA seropositivity at delivery (OR = 0.16, 95% CI = 0.07-0.37), and a severe flair in maternal alanine aminotransferase (ALT) levels (OR = 0.43, 95% CI = 0.19-0.95) during pregnancy. In addition, TDF showed more improvement in HBV DNA suppression at delivery (OR = 254.46, 95% CI = 28.39-2280.79). No significant differences were found in HBeAg seroconversion or ALT normalization; or in rates of cesarean section, emergent cesarean section, postpartum hemorrhage, prematurity, congenital malformations, or infant death. However, TDF induced more drug-related adverse events (OR = 2.33, 95% CI = 1.39-3.89) and elevated creatine kinase (CK) (OR = 9.56, 95% CI = 1.17-78.09) than in controls. The available evidence suggests that TDF is effective and safe in preventing vertical transmission of hepatitis B in pregnancies exhibiting a high viral load.


Gene expression signature-based prognostic risk score in patients with primary central nervous system lymphoma.
Better understanding of the underlying biology of primary central nervous system lymphomas (PCNSL) is critical for the development of early detection strategies, molecular markers, and new therapeutics. This study aimed to define genes associated with survival of patients with PCNSL. Expression profiling was conducted on 32 PCNSLs. A gene classifier was developed using the random survival forests model. On the basis of this, prognosis prediction score (PPS) using immunohistochemical analysis is also developed and validated in another data set with 43 PCNSLs. We identified 23 genes in which expressions were strongly and consistently related to patient survival. A PPS was developed for overall survival (OS) using a univariate Cox model. Survival analyses using the selected 23-gene classifiers revealed a prognostic value for high-dose methotrexate (HD-MTX) and HD-MTX-containing polychemotherapy regimen-treated patients. Patients predicted to have good outcomes by the PPS showed significantly longer survival than those with poor predicted outcomes (P < 0.0001). PPS using immunohistochemical analysis is also significant in test (P = 0.0004) and validation data set (P = 0.0281). The gene-based predictor was an independent prognostic factor in a multivariate model that included clinical risk stratification (P < 0.0001). Among the genes, BRCA1 protein expressions were most strongly associated with patient survival. We have identified gene expression signatures that can accurately predict survival in patients with PCNSL. These predictive genes should be useful as molecular biomarkers and they could provide novel targets for therapeutic interventions.


Emotional and psychosocial aspects of menstrual poverty in resource-poor settings: a qualitative study of the experiences of adolescent girls in an informal settlement in Nairobi.
We introduce the concept of "menstrual poverty" to categorize the multiple deprivations relating to menstruation in resource-poor settings across the Global South, and we examine how this affects the psychological well-being of adolescent girls in an urban informal settlement in Kenya. We use qualitative data collected through 34 in-depth interviews and 18 focus group discussions with girls, women, and key informants. Menstrual poverty involved practical and psychosocial challenges affecting girls at home and at school. Its emotional impacts included anxiety, embarrassment, fear of stigma, and low mood. Further research is needed on how menstrual poverty affects girls' psychological and educational outcomes.


Electrical signs of impulse conduction in spinal motoneurons.
An analysis has been made of the electrical responses recorded on the surface and within the substance of the first sacral spinal segment when the contained motoneurons are excited by single and repeated antidromic ventral root volleys. A succession of negative deflections, designated in order of increasing latency m, i, b, d, has been found. Each of those deflections possesses some physiological property or properties to distinguish it from the remainder. Indicated by that fact is the conclusion that the successive deflections represent impulse conduction through successive parts of the motoneurons that differ in behavior, each from the others. Since the spinal cord constitutes a volume conductor the negative deflections are anteceded by a positive deflection at all points except that at which the axonal impulses first enter from the ventral root into the spinal cord. Frequently two or more negative deflections are recorded together in overlapping sequence, but for each deflection a region can be found in which the onset of that deflection marks the transition from prodromal positivity to negativity. Deflection m is characteristic of axonal spikes. Latent period is in keeping with known axonal conduction velocity. Refractory period is brief. The response represented by m is highly resistant to asphyxia. Maximal along the line of ventral root attachment and attenuating sharply therefrom, deflection m can be attributed only to axonal impulse conduction. Deflection i is encountered only within the cord, and is always associated with a deflection b. The i,b complex is recordable at loci immediately dorsal to regions from which m is recorded, and immediately ventral to points from which b is recorded in isolation from i. Except for its great sensitivity to asphyxia, deflection i has properties in common with those of m, but very different from those of b or d. To judge by properties i represents continuing axonal impulse conduction into a region, however, that is readily depolarized by asphyxia. Deflection b possesses a unique configuration in that the ascending limb is sloped progressively to the right indicating a sharp decrease in velocity of the antidromic impulses penetrating the b segment. A second antidromic volley will not conduct from i segment to b segment of the motoneurons unless separated from the first by nearly 1 msec. longer than is necessary for restimulation of axons. This value accords with somatic refractoriness determined by other means. Together with spatial considerations, the fact suggests that b represents antidromic invasion of cell bodies. Deflection d is ubiquitous, but in recordings from regions dorsal and lateral to the ventral horn, wherein an electrode is close to dendrites, but remote from other segments of motoneurons, d is the initial negative deflection. In latency d is variable to a degree that demands that it represent slow conduction through rather elongated structures. When associated with deflection b, deflection d may arise from the peak of b with the only notable discontinuity provided by the characteristically sloped rising phase of b. Deflection d records the occupation by antidromic impulses of the dendrites. Once dendrites have conducted a volley they will not again do so fully for some 120 msec. Embracing the several deflections, recorded impulse negativity in the motoneurons may endure for nearly 5 msec. When the axonal deflection m is recorded with minimal interference from somatic currents, it is followed by a reversal of sign to positivity that endures as long as impulse negativity can be traced elsewhere, demonstrating the existence of current flow from axons to somata as the latter are occupied by impulses. Note is taken of the fact that impulse conduction through motoneurons is followed by an interval, measurable to some 120 msec., during which after-currents flow. These currents denote the existence in parts of the intramedullary motoneurons of after-potentials the courses of which must differ in different parts of the neurons, otherwise nothing would be recorded. The location of sources and sinks is such as to indicate that a major fraction of the current flows between axons and somata. For approximately 45 msec. the direction of flow is from dendrites to axons. Thereafter, and for the remaining measurable duration, flow is from axons to dendrites.