Augmentation of hypoxia-induced nitric oxide generation in the rat carotid body adapted to chronic hypoxia: an involvement of constitutive and inducible nitric oxide synthases.
Acute hypoxia increases the endogenous release of nitric oxide (NO) in rat carotid body and the expression of nitric oxide synthases is modulated by chronic hypoxia. The aim of the study was to examine hypoxia-induced NO generation in rat carotid body adapted to chronic hypoxia with inspired oxygen at 10% for 4 weeks. The concentration of NO was measured electrochemically with a Pt/Nafion/Pd-IrOx/POAP modified electrode inserted into the isolated carotid body superfused with bicarbonate-buffer saline at 35 degrees C. Acute hypoxia increased the concentration of NO by 471.3+/-71.4 nM in the carotid body of chronically hypoxic (CH) rats. The amount of NO release induced by hypoxia was significantly augmented when compared with that of the normoxic control (87.6+/-15.9 nM). The hypoxia-induced NO generation was markedly attenuated by pretreatment with L- NG-nitroarginine methylester (L-NAME; 500 microM), a non-selective nitric oxide synthase (NOS) inhibitor and also by removal of extracellular calcium with the calcium chelator EGTA (5 mM). Additionally, NO generation during hypoxia was reduced by 30% in the CH carotid body treated with S-methylisothiourea (SMT; 50 microM), a specific blocker of inducible NOS (iNOS). Immunohistochemical study revealed that positive iNOS protein immunoreactivity was detected in clusters of glomus cells in the carotid bodies of CH rats, but not in the normoxic group. Thus, chronic hypoxia enhances hypoxia-induced NO generation mediated by calcium-dependent NOSs and iNOS in the carotid body. Extracellular recording of sinus nerve activity of CH carotid bodies showed that L-NAME treatment enhanced the afferent discharge in response to hypoxia, confirming that the generation of NO suppresses the activities of carotid chemoreceptors. Taken together, our results suggest that hypoxia-induced NO production increases in the rat carotid body adapted to chronic hypoxia and that constitutive and inducible NOSs are involved in the NO generation. The enhancement of NO generation may play a physiological role in blunting the hypoxic chemosensitivity during chronic hypoxia.


Computed tomography-guided cutting needle biopsy for lung nodules: when the biopsy-based benign results are real benign.
Computed tomography (CT)-guided cutting needle biopsy (CNB) is an effective diagnostic method for lung nodules (LNs). The false-negative rate of CT-guided lung biopsy is reported to be up to 16%. This study aimed to determine the predictors of true-negative results in LNs with CNB-based benign results. From January 2011 to December 2015, 96 patients with CNB-based nonspecific benign results were included in this study as the training group to detect predictors of true-negative results. From January 2016 to December 2018, an additional 57 patients were included as a validation group to test the reliability of the predictors. In the training group, a total of 96 patients underwent CT-guided CNB for 96 LNs. The CNB-based results were true negatives for 82 LNs and false negatives for 14 LNs. The negative predictive value of the CNB-based benign results was 85.4% (82/96). Univariate and multivariate logistic regression analyses revealed that CNB-based granulomatous inflammation (P = 0.013, hazard ratio = 0.110, 95% confidential interval = 0.019-0.625) was the independent predictor of true-negative results. The area under the receiver operator characteristic (ROC) curve was 0.697 (P = 0.019). In the validation group, biopsy results for 47 patients were true negative, and 10 were false negative. When the predictor was used on the validation group, the area under the ROC curve was 0.759 (P = 0.011). Most of the CNB-based benign results were true negatives, and CNB-based granulomatous inflammation could be considered a predictor of true-negative results.


Protection against tissue damage in vivo by desferrioxamine: what is its mechanism of action?
Desferrioxamine (deferoxamine) is an inhibitor of iron-dependent free radical reactions that has been used to investigate the role of such reactions in several animal model systems for human disease. In vitro, desferrioxamine is not only an iron chelator but also binds other metal ions, reacts with superoxide and hydroxyl radicals, affects eicosanoid synthesis, can act as a substrate for peroxidases and can generate a reactive nitroxide radical. However, considerations of desferrioxamine concentration in vivo suggest that its ability to inhibit iron-dependent free radical reactions is the major factor that accounts for desferrioxamine's generally-protective action in animal models of human disease.


Transannular Anti-Michael Addition: Formation of 4H-Pyrazolo[5,1-c]thiazines.
The reaction of 2-(diphenylmethylene)thietan-3-one (2) with 1,2,4,5-tetrazines (3a-c) in KOH/MeOH/THF gives 4H-pyrazolo[5,1-c]thiazines (7a-c). This no vel condensation reaction proceeds via the intermediacy of an 8-(diphenylmethylene)-2H-1,4,5-thiadiazocin-7(8H)-one (5), which undergoes a multi-step rearrangement including a rare anti-Michael addition.


Intravascular large B-cell lymphoma with diffuse FDG uptake in the lung by 18FDG-PET/CT without chest CT findings.
We report a rare case of intravascular large B-cell lymphoma (IVLBCL) with diffuse fluorodeoxyglucose (FDG) uptake in the lung by (18)FDG-positron emission tomography/computed tomography (PET/CT). CT showed nodular shadow, whereas diffuse FDG uptake in PET/CT suggested IVLBCL in the lung. A random skin biopsy provided histological evidence of IVLBCL. The patient responded well to combination chemotherapy. Only two cases of IVLBCL in which diffuse pulmonary FDG uptake was demonstrated have been reported previously. FDG-PET/CT plus random skin biopsy may be useful for the early diagnosis of IVLBCL with pulmonary involvement even without convincing radiological findings in the lung.


Adrenalectomy and corticosterone replacement differentially alter CA3 dendritic morphology and new cell survival in the adult rat hippocampus.
Plastic changes in the adult mammal hippocampus can be altered by many factors and perhaps the most well-documented is stress. Stress and elevated corticosterone levels have been shown to decrease hippocampal neurogenesis and decrease the complexity of CA3 pyramidal neurons. However, the extent of these changes in relation to low and moderately elevated levels of corticosterone has yet to be fully investigated. Therefore, the aim of the present study was to determine how low to moderately elevated circulating corticosterone levels affect dendritic morphology of CA3 pyramidal cells and hippocampal neurogenesis in adult male rats. To do this, three groups of adult male Wistar rats were used: (1) Sham-operated, (2) Adrenalectomized (ADX), and (3) ADX+corticosterone replacement. Primary results show that adrenalectomy, but not moderately elevated levels of corticosterone replacement, resulted in significant atrophy of CA3 pyramidal neurons. Interestingly, moderate corticosterone replacement resulted in significantly more surviving new cells in the dentate gyrus when compared to sham controls. This work shows that circulating levels of corticosterone differentially affect plasticity in the CA3 region and the dentate gyrus.


Conservative treatment and follow-up of vaginal Gartner's duct cysts: a case series.
In women, during embryologic development, the paired Müllerian (paramesonephric) ducts fuse distally and develop into the uterus, cervix, and upper vagina. If the Wolffian ducts persist in vestigial form, they can lead to Gartner's cysts, mainly located in the right wall of the vagina. This is one of the few studies of Gartner's cysts with a series of consecutive cases over a long period of time who were exclusively subject to clinical observation. Although Gartner's cysts are found in approximately 0.1 to 0.2% of women, controversy exists regarding the course of action to be taken. We describe the cases of four women who were 38-years old, 53-years old, 37-years old, and 49-years old at their first appointment and who were of mixed ethnicity, mixed ethnicity, black, and mixed ethnicity respectively. The follow-up of these patients ranged from 2 to 17 years. In these four cases the location of the cysts was the right wall of the vagina. Transvaginal ultrasound was the test of choice for diagnostic confirmation. In the cases presented in this study, the women were asymptomatic and chose to be observed clinically. This is the first study reporting long-term clinical observation of these lesions. This study shows that conservative treatment can be a safe option for asymptomatic patients with vaginal Gartner's duct cysts.


Epilepsy management during epidemic: A preliminary observation from western China.
This study aimed to investigate whether the proposed model could manage patients with epilepsy (PWEs) during the coronavirus disease 2019 (COVID-19) outbreak. We used a model to manage the PWEs during the outbreak. Questionnaire survey and hospital data were used to explore whether PWEs under our management were affected by the virus. A total of 118 (78.7%) PWEs completed the survey. During the "model period," 22.9% (27/118) of the respondents reported antiepileptic drug (AEDs) discontinuity, including six (22.2%) PWEs who failed to purchase AEDs. Of the patients, 40.7% (22/54) failed to attend ordinary clinic, which was higher than that during the "period before model" (7.9%, 5/63). The common causes were movement limits (77.3%) and appointment failure (54.5%). A shift from ordinary clinic toward remote consultation was observed. Of the PWEs, 15.7% (13/83) referred to online pharmacy. 87.5% (14/16) of emergencies related to epilepsy were timely treated. 48.3%of PWEs thought that the epidemic had an impact on accessing medical services. Hospital data indicated that a decline in ordinary clinic visit, inpatient, surgery, and emergency attendance was observed in January and February 2020 and an increase in March 2020, as the epidemic mitigated. By contrast, online clinic visit soared in February, when the outbreak hit hard. In addition, we found no cross-infection of COVID-19 in our hospital and respondents. We demonstrated a much-needed model to manage the PWEs during the outbreak. We believed that the core architecture of this model was suitable for the management of other chronic diseases.


Relationship between blood lead concentration and nutritional status among Malay primary school children in Kuala Lumpur, Malaysia.
A cross-sectional study was conducted to identify the relationship between blood lead concentration and nutritional status among primary school children in Kuala Lumpur. A total of 225 Malay students, 113 male and 112 female, aged 6.3 to 9.8 were selected through a stratified random sampling method. The random blood samples were collected and blood lead concentration was measured by a Graphite Furnace Atomic Absorption Spectrophotometer. The nutrient intake was determined by the 24-hour Dietary Recall method and Food Frequency Questionnaire. An anthropometric assessment was reported according to growth indices (z-scores of weight-for-age, height-for-age, and weight-for-height). The mean blood lead concentration was low (3.4 +/- 1.91 ug/dL) and was significantly different between gender. Only 14.7% of the respondents fulfilled the daily energy requirement. The protein and iron intakes were adequate for a majority of the children. However, 34.7% of the total children showed inadequate intake of calcium. The energy, protein, fat and carbohydrate intakes were significantly different by gender, that is, males had better intake than females. Majority of respondents had normal mean z-score of growth indices. Ten percent of the respondents were underweight, 2.8% wasted and 5.4% stunted. Multiple linear regression showed inverse significant relationships between blood lead concentration with children's age (beta = -0.647, p < 0.001) and per capita income (beta = -0.001, p = 0.018). There were inverse significant relationships between blood lead concentration with children's age (beta = -0.877, p = 0.001) and calcium intake (beta = -0.011, p = 0.014) and positive significant relationship with weight-for-height (beta = 0.326, p = 0.041) among those with inadequate calcium intake. Among children with inadequate energy intake, children's age (beta = -0.621, p < 0.001), per capita income (beta = -0.001, p = 0.025) and protein intake (beta = -0.019, p = 0.027) were inversely and significantly related with blood lead concentration. In conclusion, nutritional status might affect the children's absorption of lead and further investigation is required for confirmation.


E3 ubiquitin-protein ligase 2 inhibits cell proliferation, migration, and invasion of non-small cell lung cancer through ubiquitination of Notch1.
This study aimed to explore the role of MIB2 in non-small cell lung cancer (NSCLC) and the underlying mechanism. Quantitative real-time PCR (QRT-PCR) and western blot were first performed to detect MIB2 expression in tumor tissues obtained from NSCLC patients (n = 30) and NSCLC cells, respectively. 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) and transwell assays were then used to examine the effect of MIB2 on the proliferation, migration and invasion of NSCLC cells. Western blot was further performed to examine the effect of Mind bomb 2 (MIB2), an E3 ligase on Notch1 protein and its ubiquitination. MIB2 was significantly down-regulated in NSCLC tissues and cells, both in mRNA and protein level. MIB2 also note worthily inhibited the proliferation, migration, and invasion of NSCLC cells. Furthermore, MIB2 only down-regulated Notch1 protein level, while facilitated the ubiquitination of Notch1. Additionally, Notch1 significantly relieved the repressed proliferation, migration and invasion of NSCLC cells induced by MIB2. Conclusively, MIB2 inhibited cell proliferation, migration and invasion via inducing Notch1 ubiquitination and degradation in NSCLC.


Attitudes of neonatal clinicians towards resuscitation of the extremely premature infant: an exploratory survey.
This study aims to explore the current attitudes of Australian neonatologists and nurses towards the resuscitation of extremely preterm infants. An anonymous questionnaire regarding resuscitation of infants of less than 28 weeks gestation was sent to all neonatologists and three registered nurses in each perinatal centre in Australia. One hundred and thirty-three questionnaires were sent. A return rate of 93% and 73% was obtained from neonatologists and nurses, respectively. Twenty-two per cent of neonatologists would 'occasionally' resuscitate at 22 weeks while none of the nurses would. A considerable proportion of neonatologists (23%) but only a few nurses (6%) would 'quite often' resuscitate 23-week infants. The majority of neonatologists (85%) and nurses (88%) would 'almost always' resuscitate 24 week infants. More than half of the respondents would 'occasionally' resuscitate a 400-499 g infant and most would resuscitate infants weighing more than 500 g. Clinicians were not as optimistic of long-term outcome as they were for survival. Typically, only 52% of neonatologists and 38% of nurses thought babies of 25 weeks gestation had a greater than 50% chance of survival without major handicap. Parental wishes and the presence of congenital abnormalities were major influences on decision to resuscitate. Hypothetically, most respondents, more doctors than nurses, would consider initiating resuscitation without parental consent at a median gestation of 25 weeks. The majority of Australian clinicians would resuscitate at a gestation of 24 weeks or greater or at a birth weight of over 500 g despite conservative estimates of intact survival. This survey has brought to light the importance of communication with parents prior to extreme premature birth.


Consenting and Assenting to Psychoanalytic Work.
The moment is opportune for a renewed look at what we understand about patient consent to treatment. Until recently, little reference to informed consent could be found in the literature, as though it has never been a preoccupation for psychoanalytic practitioners. Yet several post-Freudian authors offer reasons to suppose the risk of misunderstandings about consent. In fact, the very discovery of transference, replete with unrequited infantile wishes, implies that at some level, at some moment, in every psychoanalytic treatment there will be moments when "consent" will to some extent vacillate. A distinction, justifiable on etymological and intersubjective grounds, is made between patients' consent as a cognitive, somewhat passive, acceptance and patients' assent as an arduous, conflicted, partial disagreement with the symbolically limiting details of analytic work. It is in the discovery and working through of unexpected unconscious responses to aspects of the analytic setting and to the analyst that patients become "informed" of the unique risks to their psychic equilibrium the process poses, as well as its benefits. Instead of a static and unitary contractual event, informed consent in psychoanalysis is more properly conceived as a multilayered, repetitively posed, and necessarily ambivalent process of good-enough assenting over time.


Disability and lesion load in MS: a reassessment with MS functional composite score and 3D fast FLAIR.
The correlation between T2 lesion load (LL) detected on conventional MR sequences and disability assessed by the Expanded Disability Status Scale (EDSS) in multiple sclerosis (MS) is, at best, moderate. The present study was performed to determine whether the correlation between disability and LL is improved by (i) use of the more sensitive three-dimensional fast fluid-attenuated inversion recovery (3D-fast FLAIR) sequence; (ii) application of the newly devised MS functional composite (MSFC) score. Forty-one patients with clinically definite or probable MS were studied. All had been followed prospectively for 14 years since their first symptom. EDSS and MSFC score were assessed. Imaging was performed with 3D-fFLAIR, fast spin-echo (FSE), and T1 weighted SE sequences. 3D-fFLAIR detected a 34% higher LL than FSE, but did not improve the correlation between T2 LL and EDSS (r = 0.51 and r = 0.53); the correlation was stronger with T1 LL (r = 0.57). The MSFC score did not significantly correlate with T2 LL or T1 LL. The robust correlation in this cohort between T2 LL and EDSS may reflect the homogeneous disease duration and the wide spread of disability. This correlation was not improved by the increased detection of T2 lesions with the 3D-fFLAIR sequence. The stronger correlation between T1 LL with EDSS is consistent with the finding that this subgroup of lesions represent areas of more severe tissue damage. The MSFC score did not improve the clinico-radiological paradox that is found in MS, despite including information on cognitive function.


Abiotrophia defectiva adhere to saliva-coated hydroxyapatite beads via interactions between salivary proline-rich-proteins and bacterial glyceraldehyde-3-phosphate dehydrogenase.
Abiotrophia defectiva is a species of nutritionally variant streptococci that is found in human saliva and dental plaques and that has been associated with infective endocarditis. In our previous study, it was found that A. defectiva could bind specifically to saliva-coated hydroxyapatite beads (SHA). This study identified a cell surface component of A. defectiva that promotes adherence to SHA beads. The binding of A. defectiva to SHA was reduced in the presence of antibodies against human proline-rich protein (PRP); these results suggested that PRP may be a critical component mediating interactions between A. defectiva and the salivary pellicle. Two-dimensional gel electrophoresis of whole A. defectiva cells followed by Far-Western blotting was conducted by probing with synthetic peptides analogous to the binding region of PRP known as PRP-C. The results indicate that an A. defectiva protein of 37 kDa interacts with PRP-C. The results of amino-terminal sequencing of the adhesive A. defectiva protein revealed significant similarity to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Recombinant GAPDH bound to immobilized PRP-C in a dose-dependent manner and binding of A. defectiva to SHA or to PRP was reduced in the presence of anti-GAPDH antiserum. Western blotting or electron immunomicroscopic observations with anti-GAPDH antiserum revealed that this protein was expressed in both cytosolic and cell wall fractions. These results suggest that A. defectiva could specifically bind to PRP via interactions with cell surface GAPDH; the findings suggest a mechanism underlying A. defectiva-mediated adherence to saliva-coated tooth surfaces.


Development of an analytical method for the determination of polyphenolic compounds in vegetable origin samples by liquid chromatography and pulsed amperometric detection at a glassy carbon electrode.
A sensitive and accurate method for the determination of polyphenolic compounds in artichoke bract extracts and olive mill wastewaters by liquid chromatography coupled with pulsed amperometric detection at a glassy carbon working electrode was developed. Preliminary experiments were carried out by cyclic voltammetry to investigate the electrochemical behavior of polyphenols under different mobile phase compositions, and to test the detection and cleaning electrode potentials. Chromatographic separations were performed by using a core-shell C18 column, eluted with acetic acid and acetonitrile, by combined concave-linear binary gradients. Under the optimized experimental conditions, a good column efficiency and peak symmetry were observed, also for stereo and positional isomeric compounds. The developed three-step potential waveform for pulsed amperometric detection was successfully applied for the sensitive chromatographic determination of polyphenols in artichoke extracts and olive mill wastewaters. Linearity, precision and sensitivity of the proposed method have been evaluated. A wide linear range of response (up to 20 mg/L) has been obtained for all the investigated compounds. Detection and quantification limits in the vegetable origin sample extracts were in the range 0.004-0.6 mg/L and 0.01-2mg/L, respectively, while the injection-to-injection repeatability (n=6) ranged from 5 to 13%. The obtained results confirmed the excellent sensitivity of the electrochemical detection, and its suitability for the determination of electroactive polyphenolic compounds at low concentration levels.


Online assessment of risk factors for dementia and cognitive function in healthy adults.
Several potentially modifiable risk factors for cognitive decline and dementia have been identified, including low educational attainment, smoking, diabetes, physical inactivity, hypertension, midlife obesity, depression, and perceived social isolation. Managing these risk factors in late midlife and older age may help reduce the risk of dementia; however, it is unclear whether these factors also relate to cognitive performance in older individuals without dementia. Data from 14 201 non-demented individuals aged >50 years who enrolled in the online PROTECT study were used to examine the relationship between cognitive function and known modifiable risk factors for dementia. Multivariate regression analyses were conducted on 4 cognitive outcomes assessing verbal and spatial working memory, visual episodic memory, and verbal reasoning. Increasing age was associated with reduced performance across all tasks. Higher educational achievement, the presence of a close confiding relationship, and moderate alcohol intake were associated with benefits across all 4 cognitive tasks, and exercise was associated with better performance on verbal reasoning and verbal working memory tasks. A diagnosis of depression was negatively associated with performance on visual episodic memory and working memory tasks, whereas being underweight negatively affected performance on all tasks apart from verbal working memory. A history of stroke was negatively associated with verbal reasoning and working memory performance. Known modifiable risk factors for dementia are associated with cognitive performance in non-demented individuals in late midlife and older age. This provides further support for public health interventions that seek to manage these risk factors across the lifespan.


Trend in Blood Pressure Control Post Antihypertensive Drug Initiation in the U.S.
The aim of this study is to evaluate the temporal trends in systolic blood pressure control over 18 months after blood pressure‒lowering drug initiation in the U.S. From U.S. nationally representative electronic health records, 1,036,775 adults initiating and continuing blood pressure‒lowering drugs for ≥18 months during 2006-2018 were identified (January 2021). Prevalence trends of cardiovascular disease, diabetes, and depression at blood pressure‒lowering drug initiation, blood pressure‒lowering drug therapy intensification over 18 months, and the adjusted probability of achieving systolic blood pressure control 6 months after baseline and sustaining the control for over 18 months were evaluated. At blood pressure‒lowering drug initiation, the prevalence of diabetes and depression consistently increased during the study period across all age groups, particularly in those aged 18-49 years, whereas the prevalence of cardiovascular disease was stable. Adjusted probabilities of achieving sustainable systolic blood pressure control by age group were 0.62 (95% CI=0.61, 0.63) for ages 18-39 years, 0.55 (95% CI=0.55, 0.56) for ages 40-49 years, 0.50 (95% CI=0.49, 0.50) for ages 50-59 years, 0.43 (95% CI=0.42, 0.43) for ages 60-69 years, and 0.37 (95% CI=0.37, 0.38) for ages 70-80 years. Those with cardiovascular disease or cardiovascular disease and diabetes had approximately 20% lower adjusted probability of achieving systolic blood pressure control (31%/29%) than those without these conditions (52%, p<0.01). Those with depression had a 4% higher probability of systolic blood pressure control than those without the condition (49% vs 45%, p<0.01). In the U.S., only 30%-50% of the population are achieving sustainable blood pressure control over 18 months after blood pressure‒lowering drug initiation, with no indication of improvement in control over the last decade.


Validity, reliability and sensitivity of measures of sporting performance.
Performance testing is one of the most common and important measures used in sports science and physiology. Performance tests allow for a controlled simulation of sports and exercise performance for research or applied science purposes. There are three factors that contribute to a good performance test: (i) validity; (ii) reliability; and (iii) sensitivity. A valid protocol is one that resembles the performance that is being simulated as closely as possible. When investigating race-type events, the two most common protocols are time to exhaustion and time trials. Time trials have greater validity than time to exhaustion because they provide a good physiological simulation of actual performance and correlate with actual performance. Sports such as soccer are more difficult to simulate. While shuttle-running protocols such as the Loughborough Intermittent Shuttle Test may simulate physiology of soccer using time to exhaustion or distance covered, it is not a valid measure of soccer performance. There is a need to include measures of skill in such protocols. Reliability is the variation of a protocol. Research has shown that time-to-exhaustion protocols have a coefficient of variation (CV) of >10%, whereas time trials are more reliable as they have been shown to have a CV of <5%. A sensitive protocol is one that is able to detect small, but important, changes in performance. The difference between finishing first and second in a sporting event is <1%. Therefore, it is important to be able to detect small changes with performance protocols. A quantitative value of sensitivity may be accomplished through the signal : noise ratio, where the signal is the percentage improvement in performance and the noise is the CV.


MtDNA-maintenance defects: syndromes and genes.
A large group of mitochondrial disorders, ranging from early-onset pediatric encephalopathic syndromes to late-onset myopathy with chronic progressive external ophthalmoplegia (CPEOs), are inherited as Mendelian disorders characterized by disturbed mitochondrial DNA (mtDNA) maintenance. These errors of nuclear-mitochondrial intergenomic signaling may lead to mtDNA depletion, accumulation of mtDNA multiple deletions, or both, in critical tissues. The genes involved encode proteins belonging to at least three pathways: mtDNA replication and maintenance, nucleotide supply and balance, and mitochondrial dynamics and quality control. In most cases, allelic mutations in these genes may lead to profoundly different phenotypes associated with either mtDNA depletion or multiple deletions.


Strand Displacement Strategies for Biosensor Applications.
DNA has many unique properties beyond encoding genetic information, one of which is its physicochemical stability based on Watson-Crick base pairing. Differences in sequence complementarity between multiple DNA strands can lead to the strand displacement reaction (SDR). SDRs have been regularly applied in synthetic biology, drug delivery, and, importantly, biosensing. SDR-based biosensors have high controllability, high sensitivity, and low interference, and can be used for multiplexed detection. Such biosensors have been demonstrated to detect nearly every class of biomolecule. As the field continues to mature, such platforms can be used as an integral tool for the manipulation of biomolecular reactions, bringing biosensors one step closer to the ultimate goal of point-of-care systems.


Therapeutic endoscopy of the hepatobiliary and pancreatic system: a Vietnamese experience.
Therapeutic endoscopic retrograde cholangiopancreatography (ERCP) was initially utilized at Binh Dan Hospital, Viet Nam, in August 1993. From August 1993 through March 1997, 318 ERCP procedures were performed on 271 patients. It was not possible to obtain cholangiography in 32 cases of the 318 procedures of ERCP, for a success rate of diagnostic ERCP approaching 89%. Cases treated by ERCP included: 14 cases of Ascaris lumbricoides in the common bile duct (CBD). 69 cases of bile duct stones. 12 cases managed by nasobiliary catheter drainage. 3 cases treated by bile duct stent. Sphincterotomy was attempted on 108 cases. Complications included: 5 cases of acute pancreatitis. 7 cases of purulent cholangitis, which resulted in 1 death. 2 cases of retroperitoneal duodenal perforation. 9 cases of postsphincterotomy bleeding. We conclude that ERCP is a useful therapeutic modality for bile duct stones and parasitic worms in the bile ducts.


Parallels between Mammalian Mechanisms of Monoallelic Gene Expression.
Different types of monoallelic gene expression are present in mammals, some of which are highly flexible, whereas others are more rigid. These include allelic exclusion at antigen receptor loci, the expression of olfactory receptor genes, genomic imprinting, X-chromosome inactivation, and random monoallelic expression (MAE). Although these processes play diverse biological roles, and arose through different selective pressures, the underlying epigenetic mechanisms show striking resemblances. Regulatory transcriptional events are important in all systems, particularly in the specification of MAE. Combined with comparative studies between species, this suggests that the different MAE systems found in mammals may have evolved from analogous ancestral processes.


Chronic calcaneal osteomyelitis associated with soft-tissue defect could be successfully treated with platelet-rich plasma: a case report.
Chronic osteomyelitis associated with soft-tissue defect following surgical management is a severe complication for orthopaedic surgeons. Traditionally, the treatment protocol for the notorious complication involved thorough debridement, bone grafting, long-term antibiotic use and flap surgery. Alternatively, platelet-rich plasma (PRP), a high concentration of platelets collected via centrifugation, has been successfully used as an adjuvant treatment for bone and soft-tissue infection in medical practices. PRP has numerous significant advantages, including stypsis, inflammation remission and reducing the amount of infected fluid. It increases bone and soft-tissue healing and allows fewer opportunities for transplant rejection. Through many years of studies showing the advantages of PRP, it has become preferred organic product for the clinical treatment of infections, especially for chronic osteomyelitis associated with soft-tissue defect. To promote the clinical use of this simple and efficacious technique in trauma, we report the case of a patient with chronic calcaneal osteomyelitis associated with soft-tissue defect that healed uneventfully with PRP.


Emergency applications of intraosseous infusion.
Vascular access is an important step in the care of the critically ill child but can be very difficult and time consuming. Recently, intraosseous infusion has experienced a resurgence as a rapid alternative to venous cannulation. Several cases illustrate the usefulness of this technique in the emergency department. Included are the first reports of the use of intraosseous diazepam and succinylcholine.


The contribution of methamphetamine use to crime: Evidence from Australian longitudinal data.
To quantify the extent to which methamphetamine use is associated with increases in crime net of any premorbid risk of criminality among people who use the drug. Four one-month data panels from 469 participants dependent on methamphetamine were drawn from the MATES cohort (N = 501). Odds ratios for within-person effects were extracted from a random intercept logistic regression model for crime during periods of methamphetamine use compared to no use. Effects were adjusted for time-varying measures of age, other substance use, and socio-economic disadvantage (income, unemployment and unstable accommodation). Involvement in crime (property crime, drug dealing, fraud, violent crime) and days of methamphetamine in the past month were assessed using the Opiate Treatment Index. Crime was more likely during months when participants used methamphetamine compared to when they did not (OR 13.2 95% CI 8.5-20.6; AOR 4.7 95% CI 2.8-8.0), this reflecting more property crime (OR 10.6 95% CI 6.3-18.0; AOR 5.5 95% CI 2.8-10.8), violent crime (OR 8.2 95% CI 4.2-15.9; AOR 3.4 95% CI 1.5-8.0), fraud (OR 3.4, 95% CI 2.0-5.8; AOR 1.7 95% CI 0.8-3.3) and dealing drugs (OR 18.2 95% CI 10.2-32.5; AOR 5.9 95% CI 3.0-11.9), although the adjusted relationship for fraud was not significant. Effects were dose related. The use of methamphetamine was associated with significant increases in crime beyond premorbid risk for criminality. Crime is a likely social consequence of methamphetamine use and efforts are needed to reduce this impact.


Eyes-Closed Resting EEG Predicts the Learning of Alpha Down-Regulation in Neurofeedback Training.
Neurofeedback training, which enables the trainee to learn self-control of the EEG activity of interest based on online feedback, has demonstrated benefits on cognitive and behavioral performance. Nevertheless, as a core mechanism of neurofeedback, learning of EEG regulation (i.e., EEG learning) has not been well understood. Moreover, a substantial number of non-learners who fail to achieve successful EEG learning have often been reported. This study investigated the EEG learning in alpha down-regulation neurofeedback, aiming to better understand the alpha learning and to early predict learner/non-learner. Twenty-nine participants received neurofeedback training to down-regulate alpha in two days, while eight of them were identified as non-learners who failed to reduce their alpha within sessions. Through a stepwise linear discriminant analysis, a prediction model was built based on participant's eyes-closed resting EEG activities in broad frequency bands including lower alpha, theta, sigma and beta 1 measured before training, which was validated in predicting learners/non-learners. The findings would assist in the early identification of the individuals who would not likely reduce their alpha during neurofeedback.


Interferon treatment of a transplantable rat colon adenocarcinoma: importance of tumor site.
The effect of partially purified rat interferon ( RIFN ) was evaluated in CC531 colon tumor-bearing rats. Tumor CC531 is a DMH-induced, transplantable adenocarcinoma exhibiting weak immunogenicity. When small tumor fragments were implanted under the renal capsule, daily treatment with RIFN for 5 days led to a highly significant (p less than 0.001) inhibition of tumor growth, measured 7 days after implantation. Cyclic treatment with RIFN (10(5) units/kg/day, for 7 days in weeks 2, 4, 6 and 8) significantly retarded the development of artificial lung metastases, induced by intravenous (i.v.) injection of 5 X 10(5) colon tumor cells. The median survival time was 177 days in the RIFN group as compared to 116 days in the control group. Three of eight animals treated with RIFN showed no sign of lung metastases when they were killed 250 days after tumor cell injection. The same cyclic RIFN treatment which was effective in the lung metastases model had no influence on the growth of liver metastases evoked by the intraportal injection of 5 X 10(5) tumor cells. Laparotomy performed at days 30 and 50 after inoculation revealed equal numbers of liver metastases in the RIFN -treated group and the control group. The mean survival times obtained in the two groups were 96 +/- 20 days and 108 +/- 14 days, respectively (0.05 less than p less than 0.10). The results indicate that (1) there is no inherent resistance of this solid tumor to RIFN therapy and (2) the effect of RIFN treatment is determined by the site of tumor development. The finding that the liver can provide a protective environment against tumor immunity may have important clinical implications.


Advances in amplification-free detection of nucleic acid: CRISPR/Cas system as a powerful tool.
Amplification technologies such as polymerase chain reaction (PCR) play an important role in nucleic acid detection. However, they require bulky and sophisticated thermal cycling instrument, as well as are prone to get false-positive results due to amplicon contamination. Currently, CRISPR/Cas system has become an increasingly popular diagnostic tool for nucleic acid with the discovery of its trans-cleavage activity which can degrade single-stranded DNA or RNA at a very high turnover rate. This inherent signal amplification capability allows CRISPR/Cas system to detect unamplified nucleic acids. Here, we reviewed the recent advances of CRISPR-based amplification-free methods for nucleic acid detection. With the assistance of various signal enhancement strategies, the detection sensitivity could be comparable to that of amplification-based methods. We then presented the pros and cons of these methods. And the subsistent challenges including sample preparation, off-target effect, sequences limit, quantitative and multiplex detection were further discussed in this review. It is probable for CRISPR-powered detection methods to pave the road for rapid, cheap, highly sensitive and specific on-site detection without amplification.


The impacts of COVID-19 on the cruise industry based on an empirical study in China.
The coronavirus disease 2019 (COVID-19) pandemic has had an unprecedented impact on the entire cruise industry. This research aims to provide an understanding of the impacts of COVID-19 on the cruise industry from various stakeholders and recommend corresponding post-COVID recovery strategies for building a sustainable cruise industry. By conducting 22 semi-structured interviews in Shanghai, China and analysing the interview data using content analysis, this research finds five aspects of the impacts that are worth discussing, namely social, health and well-being, regulatory, operational, and financial aspects. Key findings include the impacts of different stakeholders' opinions, the problems existing in the current cruise industry, and the potential for future improvement. Recommendations and recovery strategies are proposed to mitigate the negative impacts. This research not only explores the impact of COVID-19 on cruise tourism and fosters recommendations in the most fast-developing region (China) but also facilitates researchers and policymakers to understand the effects of the pandemic and proposes future risk mitigation strategies.


Avoiding hospital readmission of a person with dementia.
People with dementia can be confused or distressed by hospital admission, which can then impact on their physical health. Aysha Mendes considers ways to reduce distress and enhance wellbeing.


5-Year Outcomes of Drug-Coated Balloons for Peripheral Artery In-Stent Restenosis, Long Lesions, and CTOs.
Long-term data on drug-coated balloon (DCB) outcomes in complex femoropopliteal atherosclerotic lesions are limited. The authors sought to report 5-year safety and effectiveness outcomes of a paclitaxel DCB for the treatment of de novo in-stent restenosis (ISR), long lesions (LL), or chronic total occlusions (CTOs) in the prespecified imaging cohorts of the IN.PACT Global Study. The IN.PACT Global study was a prospective, international single-arm study. Assessments through 5 years included freedom from clinically driven target lesion revascularization (CD-TLR), a safety composite (freedom from device- and procedure-related death to 30 days, and freedom from major target limb amputation and freedom from clinically driven target vessel revascularization within 60 months), and major adverse events. The prespecified imaging cohorts enrolled 132 de novo ISR, 158 LL, and 127 CTO participants. Kaplan-Meier estimates of freedom from CD-TLR through 5 years were 58.0% (ISR), 67.3% (LL), and 69.8% (CTO). The cumulative incidences of the composite safety endpoint were 56.0% (ISR), 65.7% (LL), and 69.8% (CTO). The 5-year freedom from all-cause mortality with vital status update were 81.4% (ISR), 75.2% (LL), and 78.2% (CTO). Within the ISR cohort, 15.9% of participants experienced 2 or more TLRs, compared with 9.5% and 5.5% in the LL and CTO groups, respectively. Results demonstrate long-term safety and effectiveness of this DCB in all 3 cohorts, with low reintervention rates in the LL and CTO cohorts and no safety issues. These results support the inclusion of this DCB into the treatment algorithm for complex femoropopliteal disease.


Spatial perturbations within an RNA promoter specifically recognized by a viral RNA-dependent RNA polymerase (RdRp) reveal that RdRp can adjust its promoter binding sites.
RNA synthesis during viral replication requires specific recognition of RNA promoters by the viral RNA-dependent RNA polymerase (RdRp). Four nucleotides (-17, -14, -13, and -11) within the brome mosaic virus (BMV) subgenomic core promoter are required for RNA synthesis by the BMV RdRp (R. W. Siegel et al., Proc. Natl. Acad. Sci. USA 94:11238-11243, 1997). The spatial requirements for these four nucleotides and the initiation (+1) cytidylate were examined in RNAs containing nucleotide insertions and deletions within the BMV subgenomic core promoter. Spatial perturbations between nucleotides -17 and -11 resulted in decreased RNA synthesis in vitro. However, synthesis was still dependent on the key nucleotides identified in the wild-type core promoter and the initiation cytidylate. In contrast, changes between nucleotides -11 and +1 had a less severe effect on RNA synthesis but resulted in RNA products initiated at alternative locations in addition to the +1 cytidylate. The results suggest a degree of flexibility in the recognition of the subgenomic promoter by the BMV RdRp and are compared with functional regions in other DNA and RNA promoters.


Fluorescence polarization assay by flow cytometry.
Fluorescence polarization measurement on cell suspensions provides a highly sensitive means for detecting subtle changes in the cells, such as occur early after lymphocyte activation or on malignant transformation. We review here the principles of fluorescence polarization, its measurement by a commercially available flow cytometer and application of such assays especially in cellular immunology.


Androgen receptors in the rat brain, anterior pituitary gland and ventral prostate gland: effects of orchidectomy and ageing.
Available high-affinity binding sites for 5 alpha-dihydrotestosterone (DHT) were measured in cytosols obtained from the amygdala, hypothalamus, anterior pituitary gland and ventral prostate gland of 12-week-old rats at various times after orchidectomy, and in the corresponding tissues of 18-month-old male rats. It is suggested that the lower affinity of the DHT binding reaction in brain and ventral prostatic cytosols after orchidectomy or ageing respectively, may explain, at least in part, the changes in the responsiveness of the tissues to androgens.


Randomized single blind trial to compare the short term post-operative outcome and cost analysis of laparoscopic versus ultrasound guided transversus abdominis plane block in patients undergoing bariatric surgery.
Laparoscopic Bariatric surgery despite being minimally invasive can cause moderate to severe pain in the immediate postoperative period. Adequate pain management remains a major challenge. Transversus Abdominis Plane (TAP) block is a regional anesthesia technique which blocks the sensory nerve supply of anterior-lateral abdominal wall. Primary: evaluate Laparoscopic versus ultrasound (USG)-guided TAP block on immediate post-operative analgesia after undergoing laparoscopic bariatric surgery. Secondary: compare cost effectiveness of Laparoscopic versus ultrasound-guided TAP block after undergoing bariatric surgery. Randomized Single blind study undertaken after sample size was calculated by (N) = 2(Zα + Z1-β)2σ2/δ2 which proposed 60 patients in each group. Block randomization was done after excluding redo/revision surgeries and patients were alloted Group I: Laparoscopic-guided TAP block & Group II: USG-guided TAP block. In both groups, Bilaterally, 20 ml (0.25%) bupivacaine was injected immediately after completion of bariatric surgery. SPSS v23 (IBM Corp.) was used for analysis. Group I (N = 61 53F/8 M) & Group II (N = 60 42F/18 M) were demographically comparable. Group I (3.58 ± 0.67) had significantly lower procedure time compared to Group II (12.47 ± 1.61) (p-Value < 0.001). First rescue analgesia was administered at 7.07 ± 2.61 h in Group I vs 7.21 ± 2.39 h in Group II (p-Value 0.659). In first 24 h rescue analgesic dose requirement in Group I was 1.29 ± 0.53 vs 1.39 ± 0.50 in Group II (p-Value 0.487). VAS scores during rest and movement till 24 h post-operative were statistically similar. Procedural cost was more in group II. Laparoscopic-guided TAP block is a safe and cost-effective approach for postoperative pain management after bariatric surgery and provides similar comparable analgesic effect as the USG-TAP block. Laparoscopic TAP is a surgeon delivered, easy to administer and significantly less time-consuming procedure which is feasible even when an ultrasound machine is not available.


Prediction of nitrofurantoin resistance among Enterobacteriaceae and mutational landscape of in vitro selected resistant Escherichia coli.
Nitrofurantoin (NIT) has long been a drug of choice in the treatment of lower urinary tract infections. Recent emergence of NIT resistant Enterobacteriaceae is a global concern. An ordinal logistic regression model based on PCR amplification patterns of five genes associated with NIT resistance (nfsA, nfsB, ribE, oqxA, and oqxB) among 100 clinical Enterobacteriaceae suggested that a combination of oqxB, nfsB, ribE, and oqxA is ideal for NIT resistance prediction. In addition, four Escherichia coli NIT-resistant mutants were in vitro generated by exposing an NIT-susceptible E. coli to varying concentrations of NIT. The in vitro selected NIT resistant mutants (NI2, NI3, NI4 and NI5) were found to have mutations resulting in frameshifts, premature/lost stop codons or failed amplification of nfsA and/or nfsB genes. The in vitro selected NI5 and the transductant colonies with reconstructed NI5 genotype exhibited reduced fitness compared to their parent strain NS30, while growth of a resistant clinical isolate (NR42) was found to be unaffected in the absence of NIT. These results emphasize the importance of strict adherence to prescribed antibiotic treatment regimens and dosage duration. If left unchecked, these resistant bacteria may thrive at sub-therapeutic concentrations of NIT and spread in the community.


Associations of body mass index with mortality in heart failure with preserved ejection fraction patients with ischemic versus non-ischemic etiology.
Obesity could paradoxically improve prognosis in patients with heart failure (HF), termed the "obesity paradox." Whether HF etiology could modify the "obesity paradox" is still controversial. In the present study, we aimed to assess the relationship between obesity and death in patients with heart failure with preserved ejection fraction (HFpEF) with non-ischemic versus ischemic etiologies. We analyzed 3,360 HFpEF patients from the TOPCAT (Treatment of Preserved Cardiac Function Heart Failure with an Aldosterone Antagonist) trial. Cox regression models were used to assess the association of obesity assessed by body mass index (BMI) with short-term and long-term death risk. Overweight and obesity were associated with a lower risk of long-term all-cause death in patients with non-ischemic HFpEF, even in those with class III obesity (adjusted HR: 0.61, 95% CI 0.38-0.97). However, in the ischemic subgroup, as obesity advanced, this paradoxical relationship was gradually attenuated and disappeared in class III obesity (adjusted HR: 0.93, 95% CI 0.56-1.57). Restricted cubic spline analyses confirmed the differential relationship of baseline BMI with risk of long-term death with a BMI higher than 30 kg/m2 in non-ischemic versus ischemic HFpEF. In the short-term follow-up, the beneficial effects of overweight and obesity on survival were consistently observed in all the BMI categories, with the nadirs of all-cause death risk at class III obesity category both in non-ischemic and ischemic subgroups. "Obesity paradox" was evident both in non-ischemic and ischemic HFpEF during short-term follow-up, even in those with class III obesity. However, the beneficial effect of class III obesity disappeared during long-term follow-up in ischemic HFpEF. [https://clinicaltrials.gov], identifier [NCT00094302].


FANCC localizes with UNC5A at neurite outgrowth and promotes neuritogenesis.
The Uncoordinated 5A (UNC5A) protein is part of a family of receptors that play roles in axonal pathfinding and cell migration. We previously showed that the Fanconi anemia C protein (FANCC) interacts with UNC5A and delays UNC5A-mediated apoptosis. FANCC is a predominantly cytoplasmic protein that has multiple functions including DNA damage signaling, oxygen radical metabolism, signal transduction, transcriptional regulation and apoptosis. Given the direct interaction between FANCC and UNC5A and that FANCC interferes with UNC5A-mediated apoptosis, we explored the possibility that FANCC might play a role in axonal-like growth processes. Here we show that FANCC and UNC5A are localized to regions of neurite outgrowth during neuronal cell differentiation. We also show that absence of FANCC is required for neurite outgrowth. In addition, FANCC seems required for UNC5A expression. Results from this study combined with our previous report suggest that FANCC plays a role in tissue development through the regulation of UNC5A-mediated functions.


Disease activity as a risk factor for venous thromboembolism in rheumatoid arthritis analysed using time-averaged DAS28CRP: a nested case-control study.
The objective of this study is to clarify the clinical features and risk factors of venous thromboembolism (VTE) in patients with rheumatoid arthritis (RA). We retrospectively reviewed the prevalence of VTE in RA patients who visited Hokkaido University Hospital from 2010 to 2019 and had more than 2 years of follow-up. To explore the risk to develop VTE, we selected 260 RA patients without VTE (non-VTE) via density sampling and identified the risk factors for VTE by multivariate logistic regression analysis. Univariate conditional logistic regression analysis showed older age (p < 0.0001, Odds Ratio [OR] 1.08, 95% Confidence Interval [CI] 1.04-1.14), increase of the body mass index (BMI) (p = 0.001, OR 1.17, 95% CI 1.06-1.31), higher prevalence of RA-associated lung disease (p = 0.002, OR 2.10, 95% CI 1.33-3.30) and more frequent glucocorticoid usage (p = 0.001, OR 2.09, 95% CI 1.34-3.51) in RA patients was associated with the development of VTE significantly. Furthermore, patients with higher time-averaged disease activity score 28 (DAS28) CRP were at elevated risk (p < 0.0001, OR 3.25, 95% CI 1.94-6.12). In conditional multivariate logistic regression analysis, time averaged DAS28CRP was significantly associated with the development of VTE (p = 0.0001, adjusted OR 3.40, 95% CI 1.77-7.85). Disease activity was identified as a major risk factor of VTE in patients with RA, suggesting that sustained clinical remission could be beneficial for decrease the risk of VTE.


[The prevalence of disseminated sclerosis in the Carpathian region].
The authors investigated the incidence of multiple sclerosis in the Carpathian region. It is concluded on the basis of a statistical analysis that the incidence of multiple sclerosis in this region is influenced by environmental factors.


Determination of lipid mediators in breast cancer cells using lyophilization-supercritical fluid extraction online coupled with supercritical fluid chromatography-quadrupole tandem mass spectrometry.
A lyophilization-supercritical fluid extraction coupled with supercritical fluid chromatography-quadrupole tandem mass spectrometry online method was developed for the determination of lipid mediators in breast cancer cells. Supercritical fluid extraction was applied to the cell samples for the first time due to the use of lyophilization. The conditions of supercritical fluid extraction and supercritical fluid chromatography-quadrupole tandem mass spectrometry were investigated systematically. Under the optimized conditions, all the calibration curves for the lipid mediators showed good linearity (correlation coefficient > 0.99). The limits of detection and the limits of quantification were in the range of 0.190-5.36 pg and 0.560-16.2 pg, respectively. The recoveries were in the range of 70.3-125%. The relative standard deviations of the precision ranged from 1.49-18.7% and the accuracies were higher than 84%. Compared with liquid-liquid extraction coupled with liquid chromatography and tandem mass spectrometry method, the present approach reduced the manual labor and obtained higher sensitivity as well as higher extraction recoveries for all 15 lipid mediators. Finally, the online method was applied to the quantification of lipid mediators in breast cancer cells and normal mammary epithelial cells. On the basis of the results, this lyophilization-supercritical fluid extraction online coupled with supercritical fluid chromatography-quadrupole tandem mass spectrometry method showed great promise in the analysis of lipid mediators in complex biological samples.


Cyclosporin A and tacrolimus reduce T-cell polyfunctionality but not interferon-γ responses directed at cytomegalovirus.
Cytomegalovirus (CMV) -specific immunity is often estimated by the number of in vitro CMV antigen-inducible interferon-γ-positive (IFN-γ(+) ) T cells. However, recent work indicates that simultaneous production of IFN-γ, tumour necrosis factor-α (TNF-α) and interleukin-2 (IL-2) (referred to as 'polyfunctionality') is more relevant for anti-viral protection. Here, we compared polyfunctionality of CMV-specific T cells (pp65 and IE-1 proteins) in 23 solid-organ transplant patients and seven healthy controls by flow cytometry. The proportions of TNF-α(+) /IFN-γ(+) /IL-2 cells among the activated cells were significantly reduced in transplant patients but not the frequencies of IFN-γ(+) CD8(+) T cells. Immunosuppression reduces polyfunctionality, which reflects the increased infection risk in this patient group.


Ultrastructural sequence of myelin degradation. II. Wallerian degeneration in the rat femoral nerve.
Myelin degradation in Wallerian degeneration of rat femoral nerves has been studied with the electron microscope. In the initial stages, a decrease of myelin periodicity from 115 A to 88 A was noted, followed by the transformation of the myelin structure into uniformly layered lipid inclusions. 10--14 days after nerve section, most of the inclusions found represented unstructured lipid droplets or crystals. In the later stages of degeneration, numerous pleomorphic lamellar inclusions were found, some of them resembling the structure of pi and micron-granules. Lysosomal enzyme activity was found especially in pleomorphic inclusions during the late stages of myelin degradation. Normal myelin sheaths, as well as unstructured lipid droplets and crystals, were devoid of enzyme activity. The results are compared with alterations found in Wallerian degeneration of the central nervous system.


Multilayered retinoschisis associated with optic disc pit.
Recently, optical coherence tomography (OCT) analysis has contributed greatly to the detection of posterior retinoschisis associated with optic disc pits. We report an unusual case of optic disc pit maculopathy. A 37-year-old woman with a multilayered structure of posterior retinoschisis associated with a very small, shallow optic disc pit was treated. The patient came to our clinic after a few weeks of decreased vision in her right eye. Fundus examination showed a shallow macular detachment with a tiny, shallow pit at the temporal edge of the optic disc. OCT revealed an unusual multilayered structure of retinoschisis connected to the optic disc. We performed vitrectomy with induction of the posterior hyaloid separation and gas tamponade. After vitrectomy, OCT showed a marked fluid resolution in the retinoschisis, and the patient's vision improved rapidly. The multilayered separation of retinoschisis can be caused by traction of the optic disc with a tiny, shallow pit at the temporal edge of the disc.


Sequential risk of depression in children born prematurely: A nationwide population- based analysis.
Whether children born prematurely are at a high risk of depression is still unknown. The present study examined the risk of depression in children who were born prematurely, by analyzing a national cohort in Taiwan. All premature births between January 1, 2000, and December 31, 2010, by using the Taiwan National Health Insurance Research Database. A total of 21,478 preterm children and 85,903 full-term children were enrolled in this study. Sex, level of urbanization of residential area, and parental occupation were considered. We included participants who received a diagnosis of depression in more than two clinical visits or were hospitalized due to depression. Preterm children had a 2.75-fold higher risk of depression than full-term children (95% confidence interval [CI] = 1.58-4.79, p < 0.001). Sex was not likely to be associated with depression in this study (p = 0.95). The lowest level of urbanization significantly contributed to the risk of depression in preterm children (adjusted hazard ratio = 6.8, 95% CI = 1.63-28.46, p < 0.01). Regarding parental occupation, preterm children whose parents had blue-collar and other occupations had a 3.4- and 6.06-fold higher risk of depression, respectively, compared with other children (blue-collar occupations: 95% CI = 1.04-11.15, p < 0.05; other occupations: 95% CI = 1.71-21.49, p < 0.01). Preterm children had a 2.7-fold higher risk of depression than children born full-term. Early identification, timely psychiatric care, intervention strategies, and support for their families may reduce the complications of mental illness in preterm children.


Antimicrobial susceptibility of respiratory isolates of Enterobacteriaceae and Staphylococcus aureus in Italy: incidence and trends over the period 1997-1999.
The antibiotic susceptibility of members of the family Enterobacteriaceae and of Staphylococcus aureus strains isolated from the respiratory tract was assessed over the period 1997-1999 as part of the Italian Epidemiological Observatory survey sponsored by the Smith-Kline Foundation. A standardised method was used to determine the MICs of 22 antibiotics against isolates of Klebsiella pneumoniae (n=870), Escherichia coli (n=684), Enterobacter cloacae (n=342), Enterobacter aerogenes (n=187) and Serratia marcescens (n=135) as well as the MICs of 11 antibiotics against isolates of Staphylococcus aureus (n=1,606). Overall, the susceptibility rate of Enterobacteriaceae isolates was > or = 90% to 5 agents (meropenem, imipenem, amikacin, cefepime and gentamicin); 89-80% to 2 agents (ciprofloxacin and tobramycin); and <80% to 11 agents (cefotaxime, piperacillin-tazobactam, trimethoprim-sulfamethoxazole, cefetamet, ceftriaxone, ceftazidime, aztreonam, ticarcillin-clavulanate, tetracycline, piperacillin, cefuroxime, chloramphenicol, ticarcillin, amoxicillin-clavulanate and amoxicillin). During the 3-year monitoring period, antibiotic susceptibility increased in Klebsiella pneumoniae against amoxicillin-clavulanate, in Escherichia coli against third-generation cephalosporins and aztreonam, in Enterobacter aerogenes against amoxicillin and piperacillin-tazobactam and in Serratia marcescens against most of the antibiotics. In contrast, Enterobacter cloacae showed a tendency to develop resistance to cefetamet, amikacin and ciprofloxacin. Of the total number of Staphylococcus aureus strains, 38% were methicillin resistant. Nearly 80% of the methicillin-resistant strains displayed a multiresistance pattern (additional resistance to 2 or more non-beta-lactam antibiotics). Rates of susceptibility of particular species (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus) were compared using strains from different geographical areas of Italy (northern, central and southern) and from different nosocomial areas (outpatients, intensive care unit [ICU] inpatients, non-ICU inpatients). Susceptibility of Klebsiella pneumoniae to several antibiotics was lower in southern Italy, whereas the incidence of methicillin-resistant strains was higher in northern and central Italy. The susceptibility of Escherichia coli was similar in all three areas. No significant differences in susceptibility of Klebsiella pneumoniae or Escherichia coli were found between strains from inpatients and outpatients or from inpatients admitted to ICU and non-ICU units. The incidence of methicillin-resistant Staphylococcus aureus was higher in ICU inpatients (52%) than in non-ICU inpatients (38%) and lower in outpatients (19%) than in inpatients.


Total parenteral nutrition increases serum leptin concentration in hospitalized, undernourished patients.
The hormone leptin has putative roles in both body weight homeostasis (chronic) and satiety (acute). To determine if this dual regulation is observed in hospitalized, undernourished patients, serum leptin concentration was measured before and during total parenteral nutrition (TPN) infusion. Six consecutive patients were considered undernourished, as assessed by an independent multidisciplinary nutrition team, and TPN was prescribed at an initial rate of between 5023.2 and 7333.2 kJ in the first 24 hours. Serum leptin, insulin, and glucose were measured before the infusion and at 3 and 22 hours after initiation of TPN. Baseline serum leptin concentrations correlated well with the patient's body mass index (BMI; r2 = .85, p<.05). Three hours of TPN infusion produced only modest changes in circulating leptin. However, after 22 hours, leptin concentrations increased by 1.8+/-0.5-fold (p<.05), and this increase was independent of any change in body weight. Basal leptin concentrations correlate well with BMI. TPN induces a rise in leptin concentration independent of body weight. Leptin secretion is dually regulated in hospitalized, undernourished patients.


Evaluation of spin in oncology clinical trials.
Spin, the misrepresentation of research findings, in clinical trial abstract has been shown to influence how oncologist rate a drug's efficacy. We searched PubMed for clinical trials published in ten key journals in 2017. Our primary objectives were to assess the frequency and manifestations of spin in the abstracts of those clinical trials that measured both overall survival and at least one surrogate efficacy endpoints. 124 trials were included for analysis. We found evidence of spin in 46 of 124 (37.1%, 95% CI 29.1%-45.9%) trial abstracts. Spin in the abstract results was most often due to authors emphasizing a statistically significant subgroup analysis (n = 6). Spin in the abstract conclusions was most often due to authors relying on a statistically significant surrogate endpoint to highlight the bioefficacy of the intervention (n = 17). Spin is prevalent in the abstracts of oncology clinical trials that measure OS and a surrogate endpoint. The conclusion sections of abstracts were most prone to contain spin. When OS was the primary endpoint, spin was primarily used to distract from the nonsignificant OS data. To mitigate unintentional hype for cancer therapies, we recommend authors structure their conclusions around patient-important outcomes.


Cytotoxicity testing of a dentin bonding system.
The cytotoxicity of a newly developed dentin bonding system was examined in African green monkey kidney cells and human embryonic lung cells. A tissue culture agar overlay procedure, a longitudinal study, and a cell replication assay were employed to evaluate cytotoxicity. Several of the components were discovered to be cytotoxic when tested individually in vitro. However, when used in the prescribed manner, little or no cytotoxic effect was elicited. The results of this study support further testing of these materials in vivo.


cFos is critical for MCF-7 breast cancer cell growth.
The activating protein-1 (AP-1) transcription factor is a converging point of multiple signal transduction pathways in many cells. We have previously demonstrated that overexpressing Tam67, a dominant-negative (DN) form of cJun, blocks AP-1 activity and inhibits breast cancer cell growth. We hypothesized that Tam67 forms dimers with other AP-1 proteins to suppress the growth of breast cancer cells. In the present study, we used immunoprecipitation-Western blotting to demonstrate that Tam67 binds all Jun and Fos proteins in breast cancer cells. In addition, we used two variants of the Tam67 mutant to investigate whether Jun or Fos protein was required for breast cancer cell growth. We created a Tam/Fos mutant in which the cJun dimerization domain was replaced by the cFos dimerization domain, and a Tam/Squelcher mutant in which the cJun dimerization domain was deleted. We then isolated MCF-7 cell lines that stably expressed these cJun-DN mutants under the control of an inducible promoter. Using AP-1-dependent reporter assays, we observed that Tam67 and Tam/Fos mutants inhibited AP-1 transcriptional activity, while the Tam/Squelcher mutant did not. We then determined whether Tam/Fos or Tam/Squelcher inhibited breast cell growth as well as Tam67. We found that while Tam67 repressed cell growth, neither Tam/Fos nor Tam/Squelcher mutant affected cell growth. These results indicate that Tam67 likely inactivates Fos family member proteins to suppress breast cancer cell growth. Finally, we performed antisense experiments to knock down the expression of individual family members (cJun or cFos). Our results demonstrated that antisense cFos inhibited breast cancer cell proliferation and colony formation, while antisense cJun did not. These results suggest that Tam67 suppresses breast cancer cell growth by interacting with Fos family members, specifically with cFos, to produce an inactive AP-1 complex.


Hodgkin lymphoma mimicking a large soft tissue sarcoma of the shoulder: the essential role of immunohistochemistry in histopathological diagnosis.
The shoulder and axillary regions contain various complex anatomical structures in close proximity, many of which can give rise to neoplasms. Determining the origin and hence the exact diagnosis of advanced (diffuse) tumours in this region may become problematic. In view of the tumour morphology and the affected location in this case, we highlighted the importance of Hodgkin lymphoma immunohistochemistry interpretation in a tumour which was initially suspected to be a soft tissue sarcoma.


Use of a questionnaire to obtain an alcohol history from those attending an inner city accident and emergency department.
A screening questionnaire designed to take an alcohol history was used on 996 patients attending the London Hospital Accident and Emergency Department. Questions concerned with 'binge' drinking detected many problem drinkers who were not identified by questions on weekly alcohol intake or 'CAGE' questions. The relative increase in detection was particularly marked in women.


Staining for acid-fast bacilli in surgical pathology: practice patterns and variations.
Analysis of acid-fast bacilli stains on sputum smears for the diagnosis of tuberculosis has a long history, but quality control for acid-fast bacilli in histologic sections is not as well established. In tissues, necrotizing granulomas are closely linked to positive cultures for mycobacteria. However, the practices of pathologists examining acid-fast bacilli in surgical specimens are not well described in the literature. This study characterizes practice patterns related to the histologic interpretation of acid-fast bacilli stains. A survey invitation was sent to 1299 pathologists including members of the Pulmonary Pathology Society and randomly selected fellows of the College of American Pathologists. Twenty-one questions inquired about demographics, ordering and interpreting acid-fast bacilli stains, reporting, and correlation. Of the 392 responses (30.2% response rate), 363 respondents review acid-fast bacilli stains on histologic sections. Approximately half of respondents practice in an academic setting, with the other half in community practice. Most respondents examine the entire acid-fast bacilli slide with the ×40 objective; approximately half confirm the organisms under oil immersion at ×100. There was considerable variation in when an acid-fast bacilli stain is ordered, as well as possible additional workup for negative cases, reporting of results, correlation with clinical and culture findings, and training. Many respondents reported never having been taught a general approach to acid-fast bacilli interpretation. There is substantial variation in practice patterns involving all aspects of ordering, histologic examination, and reporting of acid-fast bacilli stains. Future efforts to standardize the interpretation of acid-fast bacilli stains can potentially improve the diagnosis of mycobacterial disease.


Salt loading reduces hypothalamic noradrenaline release after noxious stimuli.
Salt loading reduces neuroendocrine responses to stressful stimuli. Noxious stimuli facilitate noradrenaline release in the hypothalamus and, as a result, activate oxytocin neurones. Here, we examined effects of salt loading upon plasma oxytocin concentrations and noradrenaline release in the hypothalamus after footshocks. Male rats were allowed to drink 2% NaCl for 7 days. Salt loading reduced the footshock-induced increase in plasma oxytocin concentrations and noradrenaline release in the supraoptic nucleus (SON). Acute administration of hypertonic saline also attenuated the footshock-induced noradrenaline increase in the supraoptic nucleus. In contrast, salt loading did not significantly change activation of A1 catecholaminergic neurones in the medulla oblongata, as measured by expression of Fos protein. These data suggest that salt loading presynaptically suppresses noradrenaline release in the hypothalamus and oxytocin release into the blood after footshocks.


A druggable pocket on PSMD10Gankyrin that can accommodate an interface peptide and doxorubicin.
PSMD10Gankyrin, a proteasomal chaperone is also an oncoprotein. Overexpression of PSMD10Gankyrin is associated with poor prognosis and survival in many cancers. Therefore, PSMD10Gankyrin is a sought-after drug target in many hard-to-treat cancers. However, its surface appears flat and undruggable. Here, we build on our earlier discovery of a common hot spot region that defined the interface of multiple interacting partners of PSMD10Gankyrin to expose vulnerable spots for a peptide and a small molecule inhibitor. High throughput virtual screening was used to screen compounds against PSMD10Gankyrin. Interaction of PSMD10Gankyrin with the drug or protein (CLIC1) or peptide was studied using any one or more of these techniques; Microscale Thermophoresis, limited trypsinolysis, SPR and ITC. Cytotoxic effect of doxorubicin was evaluated using MTT assay. We identified doxorubicin as the first-generation small molecule inhibitor of PSMD10Gankyrin. K116 and to a lesser extent R41 on PSMD10Gankyrin contribute to the bulk of binding energy for the peptide EEVD, CLIC1 and doxorubicin. We further demonstrate that PSMD10Gankyrin is an intended target for doxorubicin in cells. Drug design against protein interactions in general and PSMD10Gankyrin in particular, remains a challenge. We provide consolidated biophysical evidence for the use of a shared interface motif EEVD as a possible inhibitor of interaction network in cancers driven by PSMD10Gankyrin. We identify a chemical scaffold for designing novel inhibitors of PSMD10Gankyrin. These findings will impact the field of protein interactions in the context of disease biology/drug discovery.


Retinoic acid induces stage-specific antero-posterior transformation of rostral central nervous system.
We report a time-course analysis of the effect of retinoic acid (RA) on the development of the mouse central nervous system (CNS) from the beginning of gastrulation throughout induction and patterning of the neural tube. RA administration induces three different, stage-specific alterations of brain development, indicating perturbation of different morphogenetic steps during the establishment of a neural pattern. In particular, treatment at mid-late streak stage (7.2-7.4 days post coitum (d.p.c.)) results in early repression of Otx2 expression in the posterior neuroectoderm of the head fold and in the ventral mid line, including the prechordal plate and the rostralmost endoderm, followed by loss of forebrain morphological and molecular identities, as revealed by analysis of the expression of regionally-restricted brain genes (Otx2, Otx1, Emx2, Emx1 and Dlx1). In these embryos, reduction of the Otx2 expression domain correlates with hindbrain expansion marked by rostral extension of the Hoxb-1 expression domain. Our analysis indicates that RA interferes with the correct definition of both planar and vertical morphogenetic signals at specific developmental stages by affecting gene expression in the regions which are likely either to produce or to respond to these signals. We suggest that retinoids may contribute to early definition of head from trunk structures by selecting different sets of regulatory genes.


Single-blind, prospective, randomized study of cefmetazole and cefoxitin in the treatment of postcesarean endometritis.
The purpose of this study was to compare the clinical efficacy and safety of cefmetazole given by IV push with that of parenterally administered cefoxitin for the treatment of endometritis following cesarean delivery. In a single-blind, multicenter, prospective, randomized study, 355 patients with endometritis after cesarean delivery were enrolled and received medication. Administered was either cefmetazole sodium, 2 g by IV push over 1 min q 8 h, or cefoxitin sodium, 2 g IV q 6 h in a 2:1 ratio. The patients were followed for clinical responses and side effects. The cure rate for cefmetazole was 89% and for cefoxitin it was 79% (P = 0.006). The adverse events were similar in both groups. Cefmetazole was significantly more effective than cefoxitin in the treatment of endometritis following cesarean delivery.


Melatonin inhibitory effect on cAMP accumulation in the chick retina development.
During vertebrate neurodevelopment, neuritogenesis and synaptogenesis are modulated by intracellular cAMP rises. Melatonin, which is implicated in neuronal differentiation, mainly inhibits this pathway. Here, an investigation about the profile of this effect during the vertebrate neurodevelopment is reported. In the embryonic chick retinas at days 8, 12, 14, 16 and at 2 days post-hatched (E8, E12, E14, E16 and PH), those control embryonic retinas incubated only with the phosphodiesterase inhibitor at days corresponding to commencement of neuronal differentiation (E8, E12) and PH, presented cAMP levels inhibited by melatonin. While the cAMP accumulation stimulated by forskolin was inhibited in the embryonic retinas at all testing days. Neither the unselective antagonist N-acetyl-2-benziltryptamine (luzindole) nor the selective Mel(1b) antagonist 4-phenyl-2-propionamidotetralin (4-P-PDOT) blocked the melatonin concentration-dependent inhibitory effect on cAMP accumulation in the retinas initiating differentiation (E7-E9), suggesting a tight binding between melatonin and their receptors. However, 4-P-PDOT competitively reverted the melatonin effect on cAMP stimulated by forskolin during synaptogenesis stages. Together, the melatonin effect on cAMP levels in chick retina, which is mainly through melatonin receptors, is depending on the developmental period observed, probably taking part in the mechanisms surrounding the melatonin action on neuronal differentiation.


Multiple sclerosis typical clinical and MRI findings in a patient with HIV infection.
Multiple sclerosis (MS) is a demyelinating disease seldom included in the differential diagnosis of leukoencephalopathy in HIV-positive patients. We describe the clinical findings and laboratory results of a 43-year-old male with HIV infection and MS, and reviewed 11 more cases reported in the literature. The first episode of MS occurred either during or after the recognition of the HIV infection except in the few cases reported in 1989. There has been a very strong male predominance. Age at onset was between 30 and 40 years old. The most common clinical course was relapsing and remitting. Most of the cases had a normal CD4+ cell count, usually exceeding 500 cells/mm(3). Despite that CD4+ cell counts were invariable high, all the patients had multiple tests to rule out opportunistic infections and HIV-associated illness. The clinical suspicion of MS was only considered after ruling out other opportunistic infections and was supported with brain imaging showing multiple white matter evanescent lesions, the presence of black holes, and a high myelin basic protein titer in the CSF. MS is usually considered late in patients with HIV. A typical MS course with suggestive MRI lesions and absence of severe immune suppression should suggest the diagnosis. It is possible that as with other MS patients, earlier initiation of specific treatments for MS will prevent the high burden of the disease and disability in these patients, but stronger evidence for specific recommendations remains to be obtained.


Sphaerospora sensu stricto: taxonomy, diversity and evolution of a unique lineage of myxosporeans (Myxozoa).
Myxosporeans (Myxozoa) are eukaryotic parasites, primarily of fish, whose classification is in a state of flux as taxonomists attempt to synthesize the traditional morphology-based system with emerging DNA sequence-based phylogenies. The genus Sphaerospora Thélohan, 1892, which includes pathogenic species that cause significant impacts on fisheries and aquaculture, is one of the most polyphyletic taxa and exemplifies the current challenges facing myxozoan taxonomists. The type species, S. elegans, clusters within the Sphaerospora sensu stricto clade, members of which share similar tissue tropism and long insertions in their variable rRNA gene regions. However, other morphologically similar sphaerosporids lie in different branches of myxozoan phylogenetic trees. Herein, we significantly extend taxonomic sampling of sphaerosporids with SSU+LSU rDNA and EF-2 sequence data for 12 taxa including three representatives of the morphologically similar genus Polysporoplasma Sitjà-Bobadilla et Álvarez-Pellitero, 1995. These taxa were sampled from different vertebrate host groups, biogeographic realms and environments. Our phylogenetic analyses and statistical tests of single and concatenated datasets revealed Sphaerospora s. s. as a strongly supported monophyletic lineage, that clustered sister to the whole myxosporean clade (freshwater+marine lineages). Generally, Sphaerospora s. s. rDNA sequences (up to 3.7 kb) are the longest of all myxozoans and indeed metazoans. The sphaerosporid clade has two lineages, which have specific morphological, biological and sequence traits. Lineage A taxa (marine Sphaerospora spp.) have a single binucleate sporoplasm and shorter AT-rich rDNA inserts. Lineage B taxa (freshwater/brackish Sphaerospora spp.+marine/brackish Polysporoplasma spp.) have 2-12 uninucleate sporoplasms and longer GC-rich rDNA inserts. Lineage B has four subclades that correlate with host group and habitat; all Polysporoplasma species, including the type species, cluster together in one of these subclades. We thus suppress the genus Polysporoplasma and the family Polysporoplasmidae and emend the generic diagnosis of the genus Sphaerospora. The combination of morphological, biological and DNA sequence data applied in this study helped to elucidate an important part of the taxonomic puzzle within the phylum Myxozoa.


Retrograde Retrieval of a Novel Large Mitral Clip After Embolization Into the Left Ventricle.
We describe the successful retrieval of a novel large mitral clip, which embolized in a patient with severe secondary mitral regurgitation and left ventricular dysfunction, dilated left ventricle, and severely tethered mitral valve leaflets in the setting of a challenging anatomy for transcatheter edge-to-edge repair. The description highlights planning, technical issues, and possible adverse events of this bailout procedure. (Level of Difficulty: Intermediate.).


Posttransplant renal replacement therapy is an alarm signal for survival outcomes in pediatric liver transplantation.
The impact of renal replacement therapy (RRT) on the long-term survival outcomes of pediatric liver recipients remains controversial. A total of 224 patients aged <18 years, who underwent liver transplantation (LT), were divided into two groups: patients who underwent renal replacement therapy (RRT) (group R, n = 25, 11.2%) and those who did not (group N, n = 199, 88.8%). The posttransplant patient survival outcomes according to RRT use constituted the primary end-point. RRT was initiated preoperatively in 12 patients (48.0%) and postoperatively in 13 [early: <6 months after LT (n = 5, 20.0%) and late: ≥6 months after LT (n = 8, 32.0%)]. The indications for RRT included liver disease involving the kidney (44.0%) and hepatorenal syndrome (56.0%). The age at the time of LT (71.6 vs. 19.1 months) was higher, the pediatric end-stage liver disease score was lower (9.9 vs. 21.2), and the duration of hospitalization posttransplantation (41.0 vs. 27.0 days) was longer, while the rates of hepatic artery thrombosis (8.0% vs. 3.5%) were higher in group R (p < .05). The number of patients (60.0% vs. 93.0%; p < .001) and graft survival rates (68.0% vs. 93.0%; p < .001) were significantly lower in group R. Multivariate analysis revealed that posttransplant RRT and hepatic artery complications were risk factors for patient survival outcomes. Renal function was recovered in 7 patients (28.0%) in group R, and 9 (36.0%) eventually underwent kidney transplantation. The survival outcomes of children requiring posttransplant RRT were significantly worse than those of children, who did not undergo RRT. Physicians should pay meticulous attention to patients requiring post-LT RRT.


[Histologic and histomorphometric follow-up observations of osseointegration of corundum-blasted BMP-3 coated titanium test implants (Ti6Al4V) at orthotopic site in the giant rabbit].
To establish whether the additional coating of titanium implants with Bone Morphogenetic Protein-3 (BMP-3) might enhance osseous integration. Each of 15 cylindrical titanium test implants (Ti-6AI-4V) was coated using 230 micrograms porcine BMP-3. A further 15 implants with identical (corundium-blasted) surface served as negative controls. An uncoated and a BMP-3-coated test object were implanted into the femurs of 15 adult giant rabbits. New formation of bone around the implants was examined microscopically and histomorphometrically on postoperative days 14, 35 and 56. Coated implants revealed superior osseointegration with statistical evaluation using the t-test for matched samples showing a significantly higher volume of new bone 5 weeks after surgery. Microscopic examination revealed osseointegration with no connective tissue membrane around the surface of the implants. Our results indicate that composite metal implants are suitable carriers for BMP-3 and that improved fixation of the implants can be achieved.


Simultaneous multidimensional impacts of active learning revealed in a first implementation in the MENA region.
Enhancing science education in developing countries has been a focal point of many studies and efforts, but reform has mainly been driven by top-down approaches that often face impediments. A shift to active learning pedagogies can potentially address these challenges, but it has thus far been predominantly implemented and understood in developed countries. Thanks to the growing accessibility of open education resources and ubiquitous technologies, education reform can now be carried out from the bottom up. Here, we present the results of a two-year implementation of active learning in five core physics and astronomy courses comprising 2,145 students from the Middle East and North Africa (MENA) region. Simultaneous improvements are observed in both students' performance and their perception of the quality of learning; means improved by 9% (0.5 SD) and 25% (1.5 SD), respectively. The performance gap between students in the bottom quartile and those in the top quartiles was narrowed by 17%. The failure rate was reduced to a third of that in traditional classes; this is 36% better than the results in developed countries, indicating a greater need for active pedagogies by MENA students. Our findings reveal a multidimensional positive influence of active learning, the viability of its grassroots implementation with open resources, and its sustainability and reproducibility. We suggest that wider implementation can boost education-driven economic growth by 1% in per capita gross domestic product [GDP], substantially cut costs of repeating courses, and produce a more competent STEM workforce-all of which are urgently needed to stimulate development and growth.


High frequency one-step gene replacement in Trichoderma reesei. II. Effects of deletions of individual cellulase genes.
Four cellulase genes of Trichoderma reesei, cbh1, cbh2, egl1 and egl2, have been replaced by the amdS marker gene. When linear DNA fragments and flanking regions of the corresponding cellulase locus of more than 1 kb were used, the replacement frequencies were high, ranging from 32 to 52%. Deletion of the major cellobiohydrolase 1 gene led to a 2-fold increase in the production of cellobiohydrolase II; however, replacement of the cbh2 gene did not affect the final cellulase levels and deletion of egl1 or egl2 slightly increased production of both cellobiohydrolases. Based on our results, endoglucanase II accounts for most of the endoglucanase activity produced by the hypercellulolytic host strain. Furthermore, loss of the egl2 gene causes a significant drop in the filter paper-hydrolysing activity, indicating that endoglucanase II has an important role in the total hydrolysis of cellulose.


Precursor of beta-lactamase is enzymatically inactive. Accumulation of the preprotein in Saccharomyces cerevisiae.
Synthesis and properties of the bacterial precursor of beta-lactamase (E.C.18.104.22.168) were studied in Saccharomyces cerevisiae transformants. A protease-deficient yeast mutant was transformed with the plasmid pADH040-2 conferring high expression of the bla gene. Besides precisely processed beta-lactamase, transformed yeast cells contained mainly bla precursor up to the amount of 2% of total cellular protein. The precursor was shown to be synthesized on free polysomes in vivo but could be processed with rough microsomal membranes in a cell-free translation system. By applying an isolation procedure using high-salt conditions, the labile precursor could be separated in a native form from the mature beta-lactamase. Thereby it could be shown that the pre-beta-lactamase had virtually no enzymatic activity in contrast to the mature enzyme, which was indistinguishable from bacterial beta-lactamase. Furthermore, the precursor was highly susceptible to proteolytic degradation by trypsin under conditions which did not affect the mature enzyme. Accordingly, the protein conformation of the precursor must be substantially different from that of the authentic beta-lactamase, demonstrating that specific processing and transport of beta-lactamase is associated with directing the protein to a distinct conformation.


High Open-Circuit Voltage Cs2 AgBiBr6 Carbon-Based Perovskite Solar Cells via Green Processing of Ultrasonic Spray-Coated Carbon Electrodes from Waste Tire Sources.
Invited for this month cover is the group of Teresa Gatti at the Justus Liebig University (JLU) in Giessen, Germany, the group of Federico Bella at Politecnico di Torino (POLITO), Italy, and the group of Francesco Lamberti at the University of Padova (UNIPD), also in Italy. The image shows how waste tires can be converted in a conductive carbon powder that undergoes a green processing step to produce carbon electrodes for lead-free perovskite solar cells. Similar devices can be employed to harvest indoor light in order to power the Internet of Things (IoT) ecosystem. The Research Article itself is available at 10.1002/cssc.202201590.


Consolidative Radiation in DLBCL: Evidence-Based Recommendations.
Radiation therapy (RT) has been described as the most effective single agent in the treatment of lymphoma; however, contemporary lymphoma treatment rarely relies on single agents. In the modern era, the selection of appropriate patients for combined modality therapy has become increasingly complex over the last decade with the transition to immunochemotherapy, the emergence of functional imaging for response evaluation, and the improvement in conformal avoidance of normal tissues when delivering RT. Recent evidence demonstrates that selected patients with DLBCL have significantly better outcomes when RT is added to immunochemotherapy; however, there are important knowledge gaps regarding the use of functional imaging to facilitate treatment selection. This article will review the current evidence regarding the optimal use of combined modality therapy for DLBCL.


Imaging congenital anomalies of the ileum in adults:a pictorial review.
The ileal loops are anatomical location for the majority of congenital anomalies affecting the gastrointestinal tract. These include Meckel's diverticulum, ileal duplication, dysgenesis, atresia, mucosal diaphragm, and malposition of the ileum. Symptomatic lesions that often present with abdominal pain, intestinal obstruction or bleeding are usually diagnosed and treated during infancy and childhood. However, many of these congenital conditions may remain clinically silent and detected incidentally in adults undergoing radiological evaluation for unrelated medical reasons. This article presents the spectrum of the congenital ileal anomalies and their distinct features on small bowel examination and CT of the abdomen.


Synphilin-1 is developmentally localized to synaptic terminals, and its association with synaptic vesicles is modulated by alpha-synuclein.
Alpha-synuclein is the major component of Lewy bodies in patients with Parkinson's disease, and mutations in the alpha-synuclein gene are responsible for some familial forms of the disease. alpha-Synuclein is enriched in the presynapse, but its synaptic targets are unknown. Synphilin-1 associates in vivo with alpha-synuclein promoting the formation of intracellular inclusions. Additionally synphilin-1 has been found to be an intrinsic component of Lewy bodies in patients with Parkinson's disease. To understand the role of synphilin-1 in Parkinson's disease, we sought to define its localization and function in the brain. We now report that, like alpha-synuclein, synphilin-1 was enriched in neurons. In young rats, synphilin-1 was prominent in neuronal cell bodies but gradually migrated to neuropil during development. Immunoelectron microscopy of adult rat cerebral cortex demonstrated that synphilin-1 was highly enriched in presynaptic nerve terminals. Synphilin-1 co-immunoprecipitated with synaptic vesicles, indicating a strong association with these structures. In vitro binding experiments demonstrated that the N terminus of synphilin-1 robustly associated with synaptic vesicles and that this association was resistant to high salt washing but was abolished by inclusion of alpha-synuclein in the incubation medium. Our data indicated that synphilin-1 is a synaptic partner of alpha-synuclein, and it may mediate synaptic roles attributed to alpha-synuclein.


Multi-locus variable number tandem repeat analysis for Clostridium botulinum type B isolates in Japan: comparison with other isolates and genotyping methods.
Clostridium botulinum produces botulinum neurotoxin (BoNT) and causes botulism in humans and animals. Recently, 15-loci multi-locus variable number tandem repeat analysis (MLVA) for C. botulinum was developed for high-resolution and inter-lab comparative genotyping. This study examines the relation between MLVA and other genotyping methods such as pulsed-field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), BoNT/B subtyping and bont/b gene location to evaluate MLVA as a method applicable to the genetic markers for C. botulinum type B. Japanese isolates were genotyped using MLVA and were compared with strains from other sources reported previously. Results show that the discriminatory power of MLVA was comparable to that of PFGE and higher than that of MLST. The topology of the minimum spanning tree (MST) constructed using MLVA data was very consistent with the phylogenetic classifications of PFGE and MLST. The MST topology also represented genetic diversity between the strains possessing bont/b gene on chromosomes and plasmids. Some Japanese isolates including those associated with infant botulism were inferred to be related to isolates of Europe origin from MLVA genotyping results. The MLVA scheme used for this study is apparently useful not only for high-resolution molecular typing, but also for phylogenetic characterization of C. botulinum type B.


Partial displacement of histone H1 from chromatin is required before it can be phosphorylated by mitotic H1 kinase in vitro.
The massive nonselective and reversible phosphorylation of histone H1 during mitosis is a universal phenomenon among eukaryotes. The growth-associated kinase responsible for this phosphorylation is identical to the maturation promoting factor, a key regulator of the cell cycle. Here we showed that growth-associated kinase, isolated from mitotic HeLa cells which were capable of phosphorylating HeLa H1 in vitro with high activity and mostly at the same sites phosphorylated during mitosis in vivo (assayed by two-dimensional analysis of tryptic phosphopeptides), did not significantly phosphorylate chromatin-bound or nuclear H1 in vitro. Its inability to phosphorylate chromatin-bound H1 did not change when the amount of kinase was increased or the incubation was prolonged. The resistance of chromatin-bound H1 to phosphorylation did not result from chromatin aggregation. Rapid phosphorylation of H1 in vitro, as well as in a nuclear system, was restored when NaCl concentrations were raised above 200 mM where H1:DNA interactions are weakened. At 300 mM NaCl, chromatin-bound H1 was phosphorylated in a subset of the sites observed for free H1 phosphorylated in vitro. These results suggest that active displacement of H1 from chromatin DNA may take place before H1 can be fully phosphorylated during mitosis.


Treatment of minor severe acquired haemophilia. Is there a rationale for immunoadsorption?
In Acquired Haemophilia (AH) autoantibodies against blood coagulation factors, mainly FVIII, inhibit the blood coagulation cascade. The clinical symptoms can vary from minor to severe life threatening bleedings. At present it is unclear if the intensity of the treatment needs to be adapted to the severity of the disease. The clinical data and long term outcome from 20 patients suffering from minor severe AH were summarized. Bleedings requiring no blood transfusions were defined as less severe. In case of FVIII concentration <5% an immunosuppressive treatment (IT) consisting of cyclophosphamide 1-2 mg/kg BW/d and/or prednisolone 1-2 mg/kg BW/d was initiated. IT induced complete remission (CR) in only 40% of patients (8/20) after a mean time of 133.4 d (±90.7 d). Treatment associated severe side effects occurred in all patients. 15 patients required a factor substitution therapy due to proceeding bleedings. In 7 patients a partial remission (PR) of AH could be achieved; bleedings progressed in 5 of them and they underwent successfully second line immunoadsorption-based protocol. The inhibitor titer differed statistically significant between CR and PR with a mean of 3.7 BU vs. 16 BU. 5 patients had a fatal outcome mainly due to severe disease associated co morbidities. Immunosuppressive treatment failed in nearly a half of AH patients. Mortality was with 25% still high. The majority of patients required an intense long-term IT and developed severe treatment related side effect. Immediate start of IT did not control bleeding. In consequence, less severe AH also should be treated with a more rigorous regime because the occurrence of minors bleedings at initial presentation is not a predictive of clinical outcome. An Immunoadsorption-based protocol should be considered first line or even as a salvage strategy.