Severe COVID-19: A multifaceted viral vasculopathy syndrome.
The objective of this study was to elucidate the pathophysiology that underlies severe COVID-19 by assessing the histopathology and the in situ detection of infectious SARS-CoV-2 and viral capsid proteins along with the cellular target(s) and host response from twelve autopsies. There were three key findings: 1) high copy infectious virus was limited mostly to the alveolar macrophages and endothelial cells of the septal capillaries; 2) viral spike protein without viral RNA localized to ACE2+ endothelial cells in microvessels that were most abundant in the subcutaneous fat and brain; 3) although both infectious virus and docked viral spike protein was associated with complement activation, only the endocytosed pseudovirions induced a marked up-regulation of the key COVID-19 associated proteins IL6, TNF alpha, IL1 beta, p38, IL8, and caspase 3. Importantly, this microvasculitis was associated with characteristic findings on hematoxylin and eosin examination that included endothelial degeneration and resultant basement membrane zone disruption and reduplication. It is concluded that serious COVID-19 infection has two distinct mechanisms: 1) a microangiopathy of pulmonary capillaries associated with a high infectious viral load where endothelial cell death releases pseudovirions into the circulation, and 2) the pseudovirions dock on ACE2+ endothelial cells most prevalent in the skin/subcutaneous fat and brain that activates the complement pathway/coagulation cascade resulting in a systemic procoagulant state as well as the expression of cytokines that produce the cytokine storm. The data predicts a favorable response to therapies based on either removal of circulating viral proteins and/or blunting of the endothelial-induced response.


Function and regulation of yeast hexose transporters.
Glucose, the most abundant monosaccharide in nature, is the principal carbon and energy source for nearly all cells. The first, and rate-limiting, step of glucose metabolism is its transport across the plasma membrane. In cells of many organisms glucose ensures its own efficient metabolism by serving as an environmental stimulus that regulates the quantity, types, and activity of glucose transporters, both at the transcriptional and posttranslational levels. This is most apparent in the baker's yeast Saccharomyces cerevisiae, which has 20 genes encoding known or likely glucose transporters, each of which is known or likely to have a different affinity for glucose. The expression and function of most of these HXT genes is regulated by different levels of glucose. This review focuses on the mechanisms S. cerevisiae and a few other fungal species utilize for sensing the level of glucose and transmitting this information to the nucleus to alter HXT gene expression. One mechanism represses transcription of some HXT genes when glucose levels are high and works through the Mig1 transcriptional repressor, whose function is regulated by the Snf1-Snf4 protein kinase and Reg1-Glc7 protein phosphatase. Another pathway induces HXT expression in response to glucose and employs the Rgt1 transcriptional repressor, a ubiquitin ligase protein complex (SCF(Grr1)) that regulates Rgt1 function, and two glucose sensors in the membrane (Snf3 and Rgt2) that bind glucose and generate the intracellular signal to which Rgt1 responds. These two regulatory pathways collaborate with other, less well-understood, pathways to ensure that yeast cells express the glucose transporters best suited for the amount of glucose available.


The Sixth Mass Extinction: fact, fiction or speculation?
There have been five Mass Extinction events in the history of Earth's biodiversity, all caused by dramatic but natural phenomena. It has been claimed that the Sixth Mass Extinction may be underway, this time caused entirely by humans. Although considerable evidence indicates that there is a biodiversity crisis of increasing extinctions and plummeting abundances, some do not accept that this amounts to a Sixth Mass Extinction. Often, they use the IUCN Red List to support their stance, arguing that the rate of species loss does not differ from the background rate. However, the Red List is heavily biased: almost all birds and mammals but only a minute fraction of invertebrates have been evaluated against conservation criteria. Incorporating estimates of the true number of invertebrate extinctions leads to the conclusion that the rate vastly exceeds the background rate and that we may indeed be witnessing the start of the Sixth Mass Extinction. As an example, we focus on molluscs, the second largest phylum in numbers of known species, and, extrapolating boldly, estimate that, since around AD 1500, possibly as many as 7.5-13% (150,000-260,000) of all ~2 million known species have already gone extinct, orders of magnitude greater than the 882 (0.04%) on the Red List. We review differences in extinction rates according to realms: marine species face significant threats but, although previous mass extinctions were largely defined by marine invertebrates, there is no evidence that the marine biota has reached the same crisis as the non-marine biota. Island species have suffered far greater rates than continental ones. Plants face similar conservation biases as do invertebrates, although there are hints they may have suffered lower extinction rates. There are also those who do not deny an extinction crisis but accept it as a new trajectory of evolution, because humans are part of the natural world; some even embrace it, with a desire to manipulate it for human benefit. We take issue with these stances. Humans are the only species able to manipulate the Earth on a grand scale, and they have allowed the current crisis to happen. Despite multiple conservation initiatives at various levels, most are not species oriented (certain charismatic vertebrates excepted) and specific actions to protect every living species individually are simply unfeasible because of the tyranny of numbers. As systematic biologists, we encourage the nurturing of the innate human appreciation of biodiversity, but we reaffirm the message that the biodiversity that makes our world so fascinating, beautiful and functional is vanishing unnoticed at an unprecedented rate. In the face of a mounting crisis, scientists must adopt the practices of preventive archaeology, and collect and document as many species as possible before they disappear. All this depends on reviving the venerable study of natural history and taxonomy. Denying the crisis, simply accepting it and doing nothing, or even embracing it for the ostensible benefit of humanity, are not appropriate options and pave the way for the Earth to continue on its sad trajectory towards a Sixth Mass Extinction.


Serum Galectin-3 in Hepatitis C Virus Infection Declines after Successful Virus Eradication by Direct-Acting Antiviral Therapy.
Serum galectin-3 is regarded as an inflammatory marker in patients with chronic liver diseases. Hepatitis C virus (HCV) infection is associated with higher levels of inflammatory molecules which ameliorate by efficient treatment with direct-acting antivirals (DAAs). The aim of this study was to compare serum galectin-3 levels between HCV patients before treatment with DAAs and at the time of sustained virologic response at 12 weeks post-treatment (SVR12). Hepatitis B and human immunodeficiency virus-negative HCV infected patients not treated with HCV therapies before were recruited at the University Hospital of Regensburg. Galectin-3 was measured by enzyme-linked immunosorbent assay in the serum of patients with chronic HCV infection, before treatment initializing, at four and twelve weeks after the start of DAA therapy and at SVR12. Associations of serum galectin-3 with C-reactive protein (CRP), leukocyte count and measures of liver disease severity were analyzed. Liver fibrosis was assessed by acoustic radiation force impulse, the aspartate aminotransferase/platelet ratio index, and the fibrosis-4 score. In the serum of 81 HCV patients, galectin-3 did not correlate with viral load, viral genotype, CRP, leukocyte count, or the model for end stage liver disease score. Therapy with DAAs effectively diminished viral load within four weeks in all patients. The median value of the serum galectin-3 was 3.0 (Q1:2.0, Q3:4.0) ng/ml before therapy and declined to 2.4 (Q1: 1.7, Q3: 3.4) ng/ml at SVR12 (p<0.001; paired samples of 67 patients). At SVR12, serum galectin-3 was not correlated with CRP (r=0.057, p=0.646) or leu-kocyte count (r=0.222, p=0.071) and did not change with increasing fibrosis stage. The associations between serum galectin-3 and body mass index, liver steatosis or diabetes could not be observed. Elimination of HCV by DAA treatment lowered serum galectin-3 compared to the pre-treatment levels suggesting that HCV infection causes an increase of this immune-regulatory protein.


Family Ties: The Role of Family Context in Family Health History Communication About Cancer.
Family health history about cancer is an important prevention and health promotion tool. Yet few studies have identified family context factors that promote such discussions. We explored relations among family context (cohesion, flexibility, and openness), self-efficacy, and cancer communication (gathering family history, sharing cancer risk information, and frequency) in a diverse group of women enrolled in a randomized control trial. Baseline survey data for 472 women were analyzed. The women's average age was 34 years, 59% identified as Black, 31% had graduated high school, and 75% reported a family history of any cancer. Results showed that greater family cohesion and flexibility were related to higher communication frequency and sharing cancer information. Women who reported greater self-efficacy were more likely to have gathered family history, shared cancer risk information, and communicated more frequently with relatives. Openness was not associated with communication but was related to greater family cohesion and flexibility. Adjusting for demographic variables, self-efficacy, and family cohesion significantly predicted communication frequency. Women with higher self-efficacy were also more likely to have gathered family health history about cancer and shared cancer risk information. Future research may benefit from considering family organization and self-efficacy when developing psychosocial theories that in turn inform cancer prevention interventions.


Differential expression of oxidative phosphorylation genes in patients with Alzheimer's disease: implications for early mitochondrial dysfunction and oxidative damage.
In Alzheimer's disease (AD) pathogenesis, increasing evidence implicates mitochondrial dysfunction resulting from molecular defects in oxidative phosphorylation (OXPHOS). The objective of the present study was to determine the role of mRNA expression of mitochondrial genes responsible for OXPHOS in brain specimens from early AD and definite AD patients. In the present article, using quantitative real-time polymerase chain reaction (PCR) techniques, we studied mRNA expression of 11 mitochondrial-encoded genes in early AD patients (n = 6), definite AD patients (n = 6), and control subjects (n = 6). Using immunofluorescence techniques, we determined differentially expressed mitochondrial genes NADH 15-kDa subunit (complex I), cytochrome oxidase subunit 1 (complex IV), and ATPase delta-subunit (complex V) in the brain sections of AD patients and control subjects. Our quantitative reverse transcription (RT)-PCR analysis revealed a downregulation of mitochondrial genes in complex I of OXPHOS in both early and definite AD brain specimens. Further, the decrease of mRNA fold changes was higher for subunit 1 compared to all other subunits studied, suggesting that subunit 1 is critical for OXPHOS. Contrary to the downregulation of genes in complex I, complexes III and IV showed increased mRNA expressions in the brain specimens of both early and definite AD patients, suggesting a great demand on energy production. Further, mitochondrial gene expression varied greatly across AD patients, suggesting that mitochondrial DNA defects may be responsible for the heterogeneity of the phenotype in AD patients. Our immunofluorescence analyses of cytochrome oxidase and of the ATPase delta-subunit suggest that only subpopulations of neurons are differentially expressed in AD brains. Our double-labeling immunofluorescence analyses of 8-hydroxyguanosine and of cytochrome oxidase suggest that only selective, overexpressed neurons with cytochrome oxidase undergo oxidative damage in AD brains. Based on these results, we propose that an increase in cytochrome oxidase gene expression might be the result of functional compensation by the surviving neurons or an early mitochondrial alteration related to increased oxidative damage.


[A case of Chlamydia trachomatis peritonitis mimicking tuberculous peritonitis].
Lymphocytic ascites with low serum-ascites albumin gradient (SAAG) are observed mainly in tuberculous peritonitis, peritoneal carcinomatosis, and pancreatic disease. However, pelvic inflammatory disease (PID) induced generalized peritonitis causing diffuse ascites has been rarely described. We report a 26-year old female patient, who was diagnosed as generalized peritonitis with diffuse ascites due to Chlamydia trachomatis infection. Gynecologic examination did not show the clue of PID and in the analysis of ascites, low SAAG, predominant lymphocyte count and high level of adenosine deaminase were noted. Although the best impression was tuberculous peritonitis on the base of these findings, the laparoscopic finding was consistent with PID and the PCR for C. trachomatis infection in cervical swab was positive. This case suggests that C. trachomatis peritonitis should be considered as a rare cause of low SAAG and lymphocytic ascites in sexually active women and should be intensively evaluated including laparoscopic examination.


Clinical evaluation of a resin-modified glass-ionomer liner for cervical dentin hypersensitivity treatment.
To evaluate the effectiveness of two agents for treating cervical dentin sensitivity associated with gingival recession or noncarious cervical lesions. 44 patients with at least mild sensitivity affecting cervical dentin were enrolled in a longitudinal randomized clinical trial. A resin-based desensitizer or an experimental glass-ionomer was assigned to treat at most two teeth from each side of the mouth. Sensitivity was assessed by tactile and cold tests, measured with a Visual Analogue Scale at baseline, after treatment, and at 1 week, 1, 3, 6, and 12 months after treatment. Other noteworthy clinical observations were recorded. Both treatments effectively reduced dentin sensitivity (mixed linear model analysis). Sensitivity score for the glass-ionomer was significantly lower than for the resin-based desensitizer after treatment and at all follow-up periods (P < 0.0001). Some overhanging margins were observed in the glass-ionomer group, which could accumulate plaque and cause gingivitis. Despite material loss from some teeth treated with the glass-ionomer, the follow-up sensitivity scores were still lower than baseline scores.


Prostatic abscess due to carbapenem-resistant K. pneumonia: A case report.
Due to the widespread use of broad-spectrum antibiotics, the morbidity of prostate abscesses (PA) has declined dramatically. However, under special circumstances, such as invasive procedures and immunosuppressive conditions, some patients are more likely to develop this disease. Here, we present the case of a 21-year-old man, diagnosed with PA, with a history of chronic steroid use and a long-term indwelling urinary catheter. The pathogen was confirmed as carbapenem-resistant Klebsiella pneumoniae, a rare bacterium. This case indicates that immunodeficiency and invasive catheter use may be risk factors for PA and opportunistic bacterial infections. A 21-year-old young man presented with sudden onset of high fever (39.7°C). The patient had a history of long-term use of steroids and long-term indwelling urinary catheter. Digital rectal examination revealed obvious swelling and tenderness of the prostate. Subsequent pelvic magnetic resonance imaging showed a high signal lesion measuring 2.1 × 2.9 × 2.8 cm with T1 enhancement and T2 enhancement. On the 8th day of hospitalization, the patient underwent a PA drainage procedure and a pus culture was conducted. Subsequent pus and urine cultures showed the presence of Klebsiella pneumoniae, which exhibited resistance to all injectable carbapenems, cephalosporins, aminoglycosides, piperacillin-tazobactam, and quinolone drugs. On the 8th day of hospitalization, the patient underwent PA drainage surgery under general anesthesia to drain the abscess and relieve obstruction. After the surgery, the patient received a 2-week treatment of doxycycline. Finally, the patient was discharged after recovery and did not experience recurrence during the 6-month follow-up period. PA is not commonly found, but some patients are more susceptible to this disease under certain host conditions. Immunodeficiency and invasive catheter use may be risk factors for PA and opportunistic bacterial infections. The use of omadacycline for the treatment of carbapenem-resistant Klebsiella pneumoniae infections appears to be effective.


Intracervical procedures and the risk of subsequent very preterm birth: a case-control study.
To investigate the relation of prior intracervical procedures with very preterm birth. A population-based case-control study. The study was conducted in Australia between 2002 and 2004. Three hundred and forty-five women having a medically indicated and 236 having a spontaneous singleton birth between 20 and 31 weeks of gestation and 796 women selected randomly from all those giving birth at ≥37 weeks of gestation. Interview data were analysed using logistic regression. Very preterm birth. Very preterm birth was significantly associated with having any intracervical procedure [adjusted odds ratio (AOR) 2.07; 95% confidence interval (CI) 1.6-2.7], in particular curettage associated with abortion (AOR 1.80; 95% CI 1.2-2.6). Assisted reproductive technology procedures were significantly associated with medically indicated very preterm birth (AOR 3.07; 95% CI 1.8-5.3) and treatments for precancerous cervical changes were significantly associated with spontaneous very preterm birth, as follows: conization/cone biopsy (AOR 3.33; 95% CI 1.8-6.2) and cauterization/ablation (AOR 2.27; 95% CI 1.4-3.8). Suction aspiration for abortion, abnormal Pap smear without treatment and abortion without instrumentation were not associated with very preterm birth. Intracervical procedures are associated with very preterm birth. Notably, curettage rather than any other procedure associated with abortion appears to be implicated in the risk. The introduction of infection during cervical procedures may be the common link with risks found. Changing clinical practice in the management of abortion and human papillomavirus vaccination may lead to lowering the risks of very preterm birth.


Ethical issues in cardiovascular risk management: Patients need nurses' support.
Involving patients in decisions on primary prevention can be questioned from an ethical perspective, due to a tension between health promotion activities and patient autonomy. A nurse-led intervention for prevention of cardiovascular diseases, including counselling (risk communication, and elements of shared decision-making and motivational interviewing) and supportive tools such as a decision aid, was implemented in primary care. The aim of this study was to evaluate the nurse-led intervention from an ethical perspective by exploring in detail the experiences of patients with the intervention, and their views on the role of both the nurse and patient. The study had a qualitative design. 18 patients who had received the intervention participated. Data were gathered by in-depth interviews. The interviews were analysed using directed content analysis. The findings revealed that patients perceived the consultations not as an infringement on their autonomy, but as supportive to risk reduction efforts they tried but found hard to realise. They specifically emphasised the role of the nurse, and appreciated the nurse's realistic advice, encouragement, and help in understanding. Patients' views on and experiences with risk management are in line with notions of relational autonomy, caring cooperation and communicative action found in the literature. We conclude that patients define the relationship with the nurse as shared work in the process of developing a healthier lifestyle.


[Entecavir in the treatment of chronic hepatitis B: multicenter randomized trials and real clinical practice].
The goal of treatment for chronic hepatitis B (CHB) is now to improve quality of life and to prevent the poor outcomes of the disease rather than to eliminate the virus from the body. This goal may be achieved via the long-term maintenance of aviremia. According to the International and Russian clinical guidelines, entecavir is the first-line drug of choice to treat patients with CHB. For almost 10 years of world clinical practice there has been evidence that entecavir has a high efficacy and a favorable safety profile in a number of randomized clinical trials and in real medical practice worldwide, in Russia in particular. For instance, the BRAVR (Baraclude Russian Analysis of Virological Response) trial of 147 CHB patients from 10 Russian cities indicated that the rate of aviremia was 85.8% (n=147), 89.9% (n=138), 89.4% (n=97), and 93.5% (n=81) at 1, 2, 3, and 4 years, respectively. In addition to its virological, immunological, and biochemical efficacies, entecavir also proved to be effective in achieving the regression of histological changes and in preventing the decompensation of cirrhosis and the development of carcinoma. The given data permit the use of entecavir for the long-term therapy of CHB with confidence.


Sensorimotor alignment effects in the learning environment and in novel environments.
Four experiments investigated the conditions contributing to sensorimotor alignment effects (i.e., the advantage for spatial judgments from imagined perspectives aligned with the body). Through virtual reality technology, participants learned object locations around a room (learning room) and made spatial judgments from imagined perspectives aligned or misaligned with their actual facing direction. Sensorimotor alignment effects were found when testing occurred in the learning room but not after walking 3 m into a neighboring (novel) room. Sensorimotor alignment effects returned after returning to the learning room or after providing participants with egocentric imagery instructions in the novel room. Additionally, visual and spatial similarities between the test and learning environments were independently sufficient to cause sensorimotor alignment effects. Memory alignment effects, independent from sensorimotor alignment effects, occurred in all testing conditions. Results are interpreted in the context of two-system spatial memory theories positing separate representations to account for sensorimotor and memory alignment effects.


Further studies on acquisition of potential infectivity for man in closely related Trypanosoma Trypanozoon brucei clones.
Twelve clones were isolated from relapsing populations in an infection initiated with a clone-derived human serum resistant Trypanozoon. Isolates were serotyped by the immunotrypanolysis test and four distinct variable antigen types (VATs) designated WITat 4, 5, 6 and 7 were identified. Contemporaneous blood incubation infectivity testing (BIIT) showed that most clones isolated in the course of infection were resistant to normal human serum while some clones isolated after series of relapses were susceptible. Four clones (G1, G2, G7 and G15) designated WITat 5, WITat 6, WITat 6 and WITat 5 respectively behaved differently in the BIIT. One of each pair was readily sensitive while the two others were highly resistant to human serum. This observation suggests that resistance to human serum might be dissociated from variable antigen type in a serodeme of Trypanosoma rhodesiense. The resistance or susceptibility to normal human serum by Trypanozoon is not immutable but is determined by the environment in which the trypanosomes live.


Selective role of vagal and nonvagal innervation in initiation and coordination of gastric and small bowel patterns of interdigestive and postprandial motility.
Our previous studies suggested that extrinsic innervation modulates upper gut motility but is not requisite for cyclic interdigestive and postprandial motility of the stomach. However, the specific role of vagal and nonvagal extrinsic innervation in the initiation, coordination, and pattern of gastric motility in dogs after denervation of the entire upper gastrointestinal tract remains unclear. The aim of this study was to determine the role of vagal and nonvagal extrinsic innervation in control of gastric motility patterns. Mongrel dogs were subjected first to extrinsic denervation (in situ neural isolation) of the stomach, small bowel, proximal colon, liver, and pancreas but specifically maintaining vagal innervation to the stomach alone. After fasting and fed motility patterns were measured with indwelling gastric and small bowel manometry catheters, the dogs underwent transthoracic truncal vagotomy (completion of total extrinsic denervation of stomach), and motility studies were repeated. Vagal integrity to the stomach and pancreas was confirmed by means of a modified Hollander test and serum pancreatic polypeptide concentrations after the injection of exogenous insulin, respectively. We found that a cyclic motility pattern (migrating motor complex) persisted during fasting in both the stomach and the small bowel and that the patterns of the stomach and the duodenum remained temporally coordinated before and after vagotomy. However, although a cyclic phase III activity persisted in the stomach after vagotomy, the number of contractions and the motility index during phase III were decreased, and the duration between groupings of contractions was increased. No differences were noted in the duration of postprandial inhibition after feeding meals before and after vagotomy. These observations support our hypothesis that the vagal nerves are not necessary for the initiation or temporal coordination of global fasting or postprandial gastroduodenal motility patterns but are involved in modulating the pattern of contractions during gastric phase III.


Inhibition of Respiration of Candida albicans by Small Molecules Increases Phagocytosis Efficacy by Macrophages.
Candida albicans adapts to various conditions in different body niches by regulating gene expression, protein synthesis, and metabolic pathways. These adaptive reactions not only allow survival but also influence the interaction with host cells, which is governed by the composition and structure of the fungal cell wall. Numerous studies had shown linkages between mitochondrial functionality, cell wall integrity and structure, and pathogenicity. Thus, we decided to inhibit single complexes of the respiratory chain of C. albicans and to analyze the resultant interaction with macrophages via their phagocytic activity. Remarkably, inhibition of the fungal bc1 complex by antimycin A increased phagocytosis, which correlated with an increased accessibility of β-glucans. To contribute to mechanistic insights, we performed metabolic studies, which highlighted significant changes in the abundance of constituents of the plasma membrane. Collectively, our results reinforce the strong linkage between fungal energy metabolism and other components of fungal physiology, which also determine the vulnerability to immune defense reactions.IMPORTANCE The yeast Candida albicans is one of the major fungal human pathogens, for which new therapeutic approaches are required. We aimed at enhancements of the phagocytosis efficacy of macrophages by targeting the cell wall structure of C. albicans, as the coverage of the β-glucan layer by mannans is one of the immune escape mechanisms of the fungus. We unambiguously show that inhibition of the fungal bc1 complex correlates with increased accessibilities of β-glucans and improved phagocytosis efficiency. Metabolic studies proved not only the known direct effects on reactive oxygen species (ROS) production and fermentative pathways but also the clear downregulation of the ergosterol pathway and upregulation of unsaturated fatty acids. The changed composition of the plasma membrane could also influence the interaction with the overlying cell wall. Thus, our work highlights the far-reaching relevance of energy metabolism, indirectly also for host-pathogen interactions, without affecting viability.


Random or selective cell death during pancreatic involution following withdrawal of raw soya flour feeding in the rat.
Feeding a diet of raw soya flour, which contains a trypsin inhibitor, results in pancreatic hypertrophy and hyperplasia, and when this diet is withdrawn rapid involution ensues. This study examines whether cells produced during the hyperplastic response to raw soya four (RSF) are selectively destroyed during involution following the withdrawal of this diet. Six week old male Wistar rats were fed RSF. At 7, 9 and 12 d after commencing the diet, during the period of maximum cell proliferation, rats were injected with 0.5 muCi/g body weight of tritiated thymidine. The rats were continued on this diet for a further 16d. By 1 mth pancreatic DNA, RNA, protein, weight and the specific activity of pancreatic DNA were all significantly greater in rats fed RSF, compared to control rats fed a standard diet. When rats fed RSF were changed to a diet free of trypsin inhibitor for 7 d, pancreatic DNA, RNA, protein and weight returned to control values, however the specific activity of DNA remained unchanged from the RSF value. This report suggests that cell death involves cells produced before and during the hyperplastic response to RSF, since there is no change in the specific activity of the organ after involution.


Adductor Spasmodic Dysphonia Improves with Bilateral Thalamic Deep Brain Stimulation: Report of 3 Cases Done Asleep and Review of Literature.
To date, there are only six published reports of adductor spasmodic dysphonia (SD) responding to awake thalamic deep brain stimulation (DBS). We retrospectively reviewed cases of Essential Tremor (ET) with SD that were seen in our center from 2012 to 2020. We further identified those that have undergone thalamic DBS, and had a blinded laryngologist rate first the audio voice recordings before and after DBS using the Unified Spasmodic Dysphonia Rating Scale (USDRS), and the video recordings last to rate the related movements and facial grimacing. We identified three cases of adductor SD with ET that had undergone bilateral ventralis intermedius (VIM) DBS under general anesthesia. All patients noted improvement of their limb and voice tremor, as well as their SD post-DBS. Although improvement of tremor was observed even with initial programming in all three, improvement of SD was noted only upon reaching higher amplitudes or wider pulse widths. Blinded voice assessments showed improvement of USDRS scores post-DBS compared to pre-DBS, and with stimulator on compared to stimulator off. We report the first three cases of SD responding favorably to bilateral VIM asleep DBS and summarize the nine cases so far of SD who have undergone thalamic DBS.


Chagas disease control: deltamethrin-treated collars reduce Triatoma infestans feeding success on dogs.
Dogs are domestic reservoir hosts of Trypanosoma cruzi, the aetiological agent of Chagas disease. Using an experimental set-up mimicking rural mud-and-thatch houses, we evaluated the effect of deltamethrin-treated dog collars on the feeding success and survival of Triatoma infestans, the main T. cruzi vector in Latin America. Seven collared and three uncollared control dogs were exposed to colonized T. infestans at day 0 (i.e. before attachment of collars), at 15 days, and then monthly for 3 months post collar attachment. Following overnight exposure to uncollared dogs, 96% (1473/1538) of bugs fed, of which 51% (746/1473) fully engorged. Feeding rates were significantly reduced on collared dogs for up to 1 month post collar attachment with the lowest rates of 91% (551/604) observed at day 30 (P<0.05). Amongst those bugs that fed, engorgement rates were significantly reduced on collared dogs throughout the trial, during which average rates were 31% (543/1768) (P<0.001). No collar effect on individual bug survival was observed. Although observed effects on feeding and engorgement were limited, the strong association between blood-feeding, blood meal size and T. cruzi transmission suggests that deltamethrin-treated dog collars could help to control canine (and possibly human) T. cruzi infection.


SARS-CoV-2 vaccination may improve anxious, insomnia and depressive symptoms among Chinese population aged 18-75 years during the COVID-19 pandemic.
Previous studies have reported significant decreases in the incidence of mental health problems following SARS-CoV-2 vaccination. However, less relevant studies are published in China. We conducted a cross-sectional study involving Chinese adults aged 18-75 years with no known psychiatric diseases. The study used data from mental health of SARS-CoV-2 vaccinated and unvaccinated participants from May 2020 to July 2021.Three standardized scales, namely, the Generalized Anxiety Disorder-7 (GAD-7) for anxious symptoms, Patient Health Questionnaire-9 (PHQ-9) for depressive symptoms and Athens Insomnia Score-8 (AIS-8) for insomnia symptoms, as well as basic demographic questions were used. The hierarchical regression method was used for multivariate logistic regression analysis to explore the effects of SARS-CoV-2 vaccination on anxious, insomnia, and depressive symptoms. The results confirmed first that vaccinated participants experienced significantly lower anxious, insomnia, and depressive symptoms scores (P < 0.001) compared with unvaccinated participants. Second that vaccinated participants had a lower prevalence of anxious, insomnia, and depressive symptoms (P < 0.001). Third, after adjusting for potential confounders, we still observed a good correlation between vaccination and a reduced risk of anxious, insomnia, and depressive symptoms. The current study showed that SARS-CoV-2 vaccination may be helpful in improving anxious, insomnia, and depressive symptoms.


The semisynthesis of analogues of cytochrome c. Modifications of arginine residues 38 and 91.
The arginine residues at positions 38 and 91 of horse cytochrome c are absolutely conserved throughout eukaryotic evolution. For studies of the functional roles of these residues, we have prepared, by semisynthetic techniques, analogues of cytochrome c in which one or the other of the arginine residues has been modified. The products of modification by adduct formation with pentane-2,4-dione were purified and extensively characterized. In biological tests, the arginine-91-modified cytochrome c showed little difference in behaviour from native horse cytochrome c. Modification of arginine-38, however, led to extensive changes in biological and chemical properties. We also prepared and tested adducts with cyclohexane-1,2-dione and camphorquinone-10-sulphonic acid. The same effects on biological properties were noted irrespective of the nature of the modifying group. We suggest reasons for the differences in sensitivity of the two sites.


Impact of comorbidity on the outcome of laryngeal squamous cancer.
Comorbidity has been shown to be a determinant in treatment selection and survival in various cancers. We have previously shown that the Adult Comorbidity Evaluation-27 index is applicable in a United Kingdom setting, and the process of comorbidity grading by retrospective notes evaluation is an accurate and reliable process. The impact of comorbidity and other factors on survival was examined retrospectively in a cohort of 180 patients with laryngeal squamous cell cancer. This study shows for the first time that the higher comorbidity burden seen in supraglottic cancers in contrast to glottic tumors might account for the poorer prognosis. This study also externally validates the composite comorbidity-TNM staging system described by Piccirillo and shows the composite system to be a better predictor of outcome than TNM staging system alone. Moderate and severe comorbidity have a greater and statistically significant impact on survival than the TNM staging system.


The UCSD shortness of breath questionnaire has longitudinal construct validity in idiopathic pulmonary fibrosis.
Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease that often causes disabling dyspnea. In IPF and other lung diseases, patient-reported outcomes (PROs)-questionnaires designed to gather information from the patient's perspective-can determine whether therapies affect dyspnea or other outcomes meaningful to patients. Before a PRO can be used confidently as an outcome measure in a longitudinal trial, studies must demonstrate the PRO's ability to capture change over time in the target population. Our goal in this study was to examine whether the UCSD Shortness of Breath Questionnaire does so in patients with IPF. We used data from the Sildenafil Trial of Exercise Performance in Idiopathic Pulmonary Fibrosis (STEP-IPF) to perform analyses that examined associations between UCSD scores and five external measures (anchors) at baseline and over time. Anchors included the Activity domain from St. George's Respiratory Questionnaire (SGRQ-A), the Physical Functioning domain from the SF-36 (SF36-PF), forced vital capacity (FVC), diffusing capacity of the lung for carbon monoxide (DLCO), and distance walked during a timed walk test (6MWD). Linear regression models were used to examine relationships between UCSD scores and anchors over time. At baseline, UCSD scores were weakly correlated with percent predicted FVC (-0.21, p = 0.005) and percent predicted DLCO (-0.20, p = 0.008), moderately correlated with 6MWD (-0.39, p < 0.0001) and strongly correlated with SGRQ-A (0.79, p < 0.0001) and SF36-PF (-0.72, p < 0.0001). Change over time in UCSD scores was associated with change in FVC (estimate = 2.54, standard error [SE] = 1.23, p = 0.04), SGRQ-A (estimate = 7.94, SE = 1.11, p < 0.0001), SF36-PF (estimate = 6.00, SE = 1.13, p < 0.0001), and 6MWD (estimate = 4.23, SE = 1.18, p = 0.0004) but not DLCO (estimate = 0.33, SE = 1.33, p = 0.80). These results support the validity of the UCSD to assess change in dyspnea over time in patients with IPF.


5-Flourouracil-induced toxicity in both male and female reproductive systems: A narrative review.
The chemotherapeutic drug 5-flourouracil (5FU) is frequently used to treat a wide range of solid malignant tumors, such as colorectal, pancreatic, gastric, breast, and head and neck cancers. Its antitumoral effects are achieved by interfering with the synthesis of RNA and DNA and by inhibiting thymidylate synthase in both malignant and non-malignant cells. Therefore, it can be responsible for severe toxicities in crucial body organs, including heart, liver, kidney, and reproductive system. Given the fact that 5FU-induced reproductive toxicity may limit the clinical application of this drug, in this study, we aimed to discuss the main locations and mechanisms of the 5FU-induced reproductive toxicity. Initially, we discussed the impact of 5FU on the male reproductive system, which leads to damage of the seminiferous epithelial cells and the development of vacuoles in Sertoli cells. Although no noticeable changes occur at the histopathological level, there is a decrease in the weight of the prostate. Additionally, 5FU causes significant abnormalities in spermatogenesis, including germ cell shedding, spermatid halo formation, polynucleated giant cells, and decreased sperm count. Finally, in females, 5FU-induced reproductive toxicity is characterized by the presence of atretic secondary and antral follicles with reduced numbers of growing follicles, ovarian weight, and maturity impairment.


A Review of Beef Production Systems for the Sustainable Use of Surplus Male Dairy-Origin Calves Within the UK.
The UK dairy herd is predominantly of the Holstein-Friesian (HF) breed, with a major emphasis placed on milk yield. Subsequently, following years of continued single-trait selection, the beef production potential of dairy bred calves has declined. Thus, male HF calves are commonly seen as a by-product of the dairy industry. Limited markets, perceived low economic value and high rearing costs mean that these surplus calves are often euthanised shortly after birth or exported to the EU for further production. Welfare concerns have been raised regarding both euthanasia and long distance transportation of these calves. Furthermore, total UK beef consumption increased by 8.5% from 2009 to 2019. Thus, in light of this growing demand, beef from the dairy herd could be better utilized within the UK. Therefore, the potential for these calves to be used in a sustainable, cost-effective beef production system with high welfare standards within the UK requires investigation. Thus, the aim of this review was to evaluate both steer and bull beef production systems, examining the impact on performance, health, welfare, and economic potential to enable a sustainable farming practice, while meeting UK market requirements. The principal conclusions from this review indicate that there is the potential for these calves to be used in UK based production systems and meet market requirements. Of the steer production systems, a 24 month system appears to achieve a balance between input costs, growth from pasture and carcass output, albeit the literature is undecided on the optimum system. The situation is similar for bull beef production systems, high input systems do achieve the greatest gain in the shortest period of time, however, these systems are not sustainable in volatile markets with fluctuating concentrate prices. Thus, again the inclusion of a grazing period, may increase the resilience of these systems. Furthermore, production systems incorporating a period at pasture are seen to have animal welfare benefits. The main welfare concern for surplus dairy bred calves is often poor colostrum management at birth. While in steer systems, consideration needs to be given to welfare regarding castration, with the negative impacts being minimized by completing this procedure soon after birth.


Fertility Intentions and Perspectives on Contraceptive Involvement Among Low-Income Men Aged 25 to 55.
Unintended pregnancy is disproportionately reported by low-income individuals in the United States, and studies of men's roles in preventing pregnancy have largely focused on adolescents and young adults. Less is known about the pregnancy-related attitudes and behaviors among men older than 24, who are involved in the majority of pregnancies ending in a birth. Between December 2015 and August 2016, in-depth interviews were conducted with 26 low-income men in Alabama who were aged 25-55, were sexually active and did not want more children. Interviews explored men's reasons for not wanting more children, their contraceptive knowledge and attitudes, and their involvement in contraceptive decision making. Transcripts were examined using content analysis to identify themes related to men's perspectives about preventing pregnancy and using contraceptives. Participants' motivations to prevent a pregnancy centered primarily on their age and financial circumstances. Most men had limited contraceptive knowledge and perceived their risk of causing a pregnancy to be low, regardless of method use. Few men engaged in decisions about contraceptive use, despite their beliefs that men and women had a shared responsibility to prevent pregnancy. Although some men were interested in vasectomy, a few were hesitant about undergoing the procedure because they might want to have children later if their life circumstances changed, and others worried that vasectomy might affect sexual performance. Some low-income adult men were uncertain about their pregnancy desires, and many lacked contraceptive knowledge that would help them avoid unwanted pregnancy. Research is needed to identify the types of programs that could effectively promote men's constructive engagement in preventing pregnancies over their reproductive life course.


Both plant and bacterial nitrate reductases contribute to nitric oxide production in Medicago truncatula nitrogen-fixing nodules.
Nitric oxide (NO) is a signaling and defense molecule of major importance in living organisms. In the model legume Medicago truncatula, NO production has been detected in the nitrogen fixation zone of the nodule, but the systems responsible for its synthesis are yet unknown and its role in symbiosis is far from being elucidated. In this work, using pharmacological and genetic approaches, we explored the enzymatic source of NO production in M. truncatula-Sinorhizobium meliloti nodules under normoxic and hypoxic conditions. When transferred from normoxia to hypoxia, nodule NO production was rapidly increased, indicating that NO production capacity is present in functioning nodules and may be promptly up-regulated in response to decreased oxygen availability. Contrary to roots and leaves, nodule NO production was stimulated by nitrate and nitrite and inhibited by tungstate, a nitrate reductase inhibitor. Nodules obtained with either plant nitrate reductase RNA interference double knockdown (MtNR1/2) or bacterial nitrate reductase-deficient (napA) and nitrite reductase-deficient (nirK) mutants, or both, exhibited reduced nitrate or nitrite reductase activities and NO production levels. Moreover, NO production in nodules was found to be inhibited by electron transfer chain inhibitors, and nodule energy state (ATP-ADP ratio) was significantly reduced when nodules were incubated in the presence of tungstate. Our data indicate that both plant and bacterial nitrate reductase and electron transfer chains are involved in NO synthesis. We propose the existence of a nitrate-NO respiration process in nodules that could play a role in the maintenance of the energy status required for nitrogen fixation under oxygen-limiting conditions.


[Single kidney derived to sigmoid for urinary tuberculosis: remarkable morpho-functional status 51 years after surgery].
To report a solitary kidney ureterosigmoidostomy (US) secondary to urinary tuberculosis which we believe has the longest follow-up in the world literature. With this purpose we performed an analysis of the experience with such a urinary diversion published in our country, which maybe has been exaggeratedly excluded during the last decades. Male patient who was diagnosed of urinary tuberculosis at the age of 17 in 1950 and underwent left nephrectomy. Due to bladder retraction he underwent cystectomy and urinary diversion to the sigmoid colon following the technique by Coffeyone year later. Follow-up for half a century has demonstrated an excellent outcome with almost no effect on renal function and urinary tract morphology, and very low incidence of infectious complications. We review cases of US with longest follow-up in the literature, finding a remarkable number of cases followed for more than 30 years. It is true they were performed almost exclusively for benign diseases. There is only one published citation in which US was indicated for bladder carcinoma and after 31 years of follow-up patient died from a nonrelated disease at the age of 85. We also review the scarce number of papers published in our country, focusing on the "Ponencia al Congreso Nacional de Urología 1983" which presented a thorough review of "urinary diversions" with the cooperation of 27 Spanish centers.


A solid-phase synthesis of N,N'-disubstituted ureas and Perhydroimidazo
An efficient method for trapping isocyanates, generated from the Curtius rearrangement, with resin-bound amines is reported. A commercially available carboxylic acid is treated with diphenylphosphoryl azide, followed by thermal rearrangement, cooling, and trapping in one pot. Cleavage from the resin gives an N, N'-disubstituted urea in excellent purity, as demonstrated with several heterocyclic and aliphatic carboxylic acids. Further utility is shown by preparing several novel perhydroimidazo[1,5-a]pyrazines.


Therapeutic strategies for residual or recurrent intracranial aneurysms after microsurgical clipping.
Therapeutic strategies for residual or recurrent aneurysm (RRA) after microsurgical clipping have not been well established. The purpose of this study was to report our retreatment experiences with previously clipped aneurysms and to demonstrate retreatment strategies for these RRAs. From 1996-2016, we treated 68 RRAs after previous clipping. Among them, 34 patients underwent microsurgical retreatment, and the other 34 underwent endovascular retreatment. Radiographic images and clinical data were reviewed retrospectively to determine the treatment efficacy, clinical outcomes, and important factors for selecting the proper treatment modality. The most common aneurysm location was the middle cerebral artery (50%) in the microsurgery group and the internal carotid artery (47.1%) in the endovascular surgery group (p = 0.001). In the microsurgery group, 16 (47.1%) patients had additional clipping without removal of previous clips, 17 (50.0%) had clipping with removal of previous clips, and 1 (2.9%) had bypass surgery with trapping. In the endovascular surgery group, 28 (82.4%) patients had simple coiling, 5 (14.7%) had stent-assisted coiling, and 1 (2.9%) had a flow diverter. Procedure-related complications during retreatment occurred in 4 (5.9%) patients. Complete obliteration was achieved in 51 (75.0%) patients (microsurgery group, 82.4% and endovascular surgery group, 67.6%; p = 0.002). In properly selected cases, treatment of RRAs could be safely performed either by microsurgery or endovascular surgery and result in a good clinical outcome with acceptable morbidity. The decision to choose the treatment modality for RRAs after clipping is not easy but should be considered to lower the risk of retreatment.


Biocontrol of Soft Rot: Confocal Microscopy Highlights Virulent Pectobacterial Communication and Its Jamming by Rhodococcal Quorum-Quenching.
Confocal laser-scanning microscopy was chosen to observe the colonization and damage caused by the soft rot Pectobacterium atrosepticum and the protection mediated by the biocontrol agent Rhodococcus erythropolis. We developed dual-color reporter strains suited for monitoring quorum-sensing and quorum-quenching activities leading to maceration or biocontrol, respectively. A constitutively expressed cyan or red fluorescent protein served as a cell tag for plant colonization, while an inducible expression reporter system based on the green fluorescent protein gene enabled the simultaneous recording of signaling molecule production, detection, or degradation. The dual-colored pathogen and biocontrol strains were used to coinoculate potato tubers. At cellular quorum, images revealed a strong pectobacterial quorum-sensing activity, especially at the plant cell walls, as well as a concomitant rhodococcal quorum-quenching response, at both the single-cell and microcolony levels. The generated biosensors appear to be promising and complementary tools useful for molecular and cellular studies of bacterial communication and interference.


S-nitrosylation of Hsp90 promotes the inhibition of its ATPase and endothelial nitric oxide synthase regulatory activities.
Nitric oxide is implicated in a variety of signaling pathways in different systems, notably in endothelial cells. Some of its effects can be exerted through covalent modifications of proteins and, among these modifications, increasing attention is being paid to S-nitrosylation as a signaling mechanism. In this work, we show by a variety of methods (ozone chemiluminescence, biotin switch, and mass spectrometry) that the molecular chaperone Hsp90 is a target of S-nitrosylation and identify a susceptible cysteine residue in the region of the C-terminal domain that interacts with endothelial nitric oxide synthase (eNOS). We also show that the modification occurs in endothelial cells when they are treated with S-nitroso-l-cysteine and when they are exposed to eNOS activators. Hsp90 ATPase activity and its positive effect on eNOS activity are both inhibited by S-nitrosylation. Together, these data suggest that S-nitrosylation may functionally regulate the general activities of Hsp90 and provide a feedback mechanism for limiting eNOS activation.


An aptamer-based new method for competitive fluorescence detection of exosomes.
Exosomes have been recognized as promising sources of biomarkers for early cancer diagnosis due to their important role in the occurrence and metastasis of cancer, and so the development of a sensitive low-cost detection method for exosomes is highly desirable. In this paper, we report a fluorescence method for the competitive detection of exosomes based on an aptamer specific to CD63 (an exosome transmembrane protein). Aptamer-modified magnetic beads were hybridized with a Cy3-labeled short sequence complementary to a region of the aptamer. In the presence of exosomes, the CD63 on the exosomes bound to the aptamer, resulting in the shedding of the short sequence into the supernatant. The quantity of the exosomes could be estimated by detecting the fluorescence intensity in the supernatant. This method could detect exosomes at a concentration as low as 1.0 × 105 particles per μL under optimal conditions, and the feasibility of the method for exosome detection in complex clinical samples was also proved using simulated serum samples. The detection cost and difficulty are significantly reduced compared to conventional methods, while ensuring sensitivity, and so this method provides a basis for subsequent exosome detection in specific cancer cells.


[Treatment of primary immune defects].
Management of immunodeficient patients includes monitoring of growth, diet, personal hygiene and life habits, including environmental factors. Major therapeutic stays are prevention, early detection and the decisive treatment of infections. Attenuated live vaccines, non-irradiated blood products and blood containing CMV antibodies should not be applied. In the case of antibody defects immunoglobulin substitution is a specific form of treatment. Enzyme replacement is a causal approach to an adenosine deaminase defect. For numerous primary immune defects, bone marrow transplantation represents the best causal therapeutic measure. In the absence of a bone marrow donor, alternative gene therapy is available for the management of severe combined immunodeficiency on the X-chromosome (X-SCID), and adenosine deaminase deficiency (ADA deficiency).


Mitosis-specific activation of LIM motif-containing protein kinase and roles of cofilin phosphorylation and dephosphorylation in mitosis.
Actin filament dynamics play a critical role in mitosis and cytokinesis. LIM motif-containing protein kinase 1 (LIMK1) regulates actin reorganization by phosphorylating and inactivating cofilin, an actin-depolymerizing and -severing protein. To examine the role of LIMK1 and cofilin during the cell cycle, we measured cell cycle-associated changes in the kinase activity of LIMK1 and in the level of cofilin phosphorylation. Using synchronized HeLa cells, we found that LIMK1 became hyperphosphorylated and activated in prometaphase and metaphase, then gradually returned to the basal level as cells entered into telophase and cytokinesis. Although Rho-associated kinase and p21-activated protein kinase phosphorylate and activate LIMK1, they are not likely to be involved in mitosis-specific activation and phosphorylation of LIMK1. Immunoblot and immunofluorescence analyses using an anti-phosphocofilin-specific antibody revealed that the level of cofilin phosphorylation, similar to levels of LIMK1 activity, increased during prometaphase and metaphase then gradually declined in telophase and cytokinesis. Ectopic expression of LIMK1 increased the level of cofilin phosphorylation throughout the cell cycle and induced the formation of multinucleate cells. These results suggest that LIMK1 is involved principally in control of mitosis-specific cofilin phosphorylation and that dephosphorylation and reactivation of cofilin at later stages of mitosis play a critical role in cytokinesis of mammalian cells.


Risk factors for comorbid psychopathology in youth with psychogenic nonepileptic seizures.
To examine the risk factors for internalizing (anxiety, depression) and posttraumatic stress (PTSD) disorders, somatization, and anxiety sensitivity (AS) in youth with psychogenic non-epileptic seizures (PNES). 55 probands with PNES and 35 siblings, aged 8-18 years, underwent a psychiatric interview, cognitive and language testing, and completed somatization and AS questionnaires. Parents provided the subjects' medical, psychiatric, family, and adversity history information. The risk factors for the probands' internalizing disorders (girls, older age of PNES onset), somatization (older age, epilepsy), and anxiety sensitivity (girls, adversities) differed from their siblings. The risk factors in the siblings, however, were similar to the general pediatric population. Proband depression was unrelated to the study's risk variables while PTSD was significantly associated with female gender and lower Full Scale IQ. Knowledge about the specificity of the risk factors for comorbid psychopathology in youth with PNES might facilitate their early identification and treatment.


Intestinal duplication in adulthood: A rare entity, difficult to diagnose.
Duplications of the alimentary tract (ATD) are rare congenital anomalies often found early in life. They may occur anywhere in the intestinal tract but the ileum is the most frequently affected site. Clinical presentation of ATD in adults is variable and because these lesions occur so infrequently they are rarely suspected. In the present report we describe a case of ileal duplication in a 61-year-old patient with Crohn's disease. Despite various radiological investigations and medical consultations, the diagnosis was only made on the surgical specimen.


Learning dynamics in public goods games.
We extend recent analyses of stochastic effects in game dynamical learning to cases of multiplayer games and to games defined on networked structures. By means of an expansion in the noise strength we consider the weak-noise limit and present an analytical computation of spectral properties of fluctuations in multiplayer public goods games. This extends existing work on two-player games. In particular we show that coherent cycles may emerge driven by noise in the adaptation dynamics. These phenomena are not too dissimilar from cyclic strategy switching observed in experiments of behavioral game theory.


[Phlebotrophic therapy with venarus in patients suffering from lower limb chronic venous insufficiency].
The article deals with medicamentous therapy of lower limb chronic venous insufficiency (LLCVI). The authors analysed herein the results of treating patients suffering from LLCVI using Venarus whose basic components are diosmin and hesperidin. The obtained findings demonstrated that clinical administration of this phlebotonic agent is effective in the given cohort of patients during the whole therapeutic course, with a maximum positive effect observed within the first 4 weeks. The parameters of quality of life of the patients with LLCVI improved significantly on the background of taking the phlebotonic Venarus.


PATIENT-RELATED FACTORS AND CLINICAL MANAGEMENT OUTCOMES OF RETINOBLASTOMA IN CALABAR, NIGERIA.
Retinoblastoma is curable in industrialized countries. However, it is associated with mortality in resource-poor nations due to disparities and poor access to eye care. Aim was to determine the relationships between patient-related factors and clinical outcomes of Retinoblastoma management in a tertiary hospital in Nigeria. This was a retrospective study of all children who were diagnosed and treated for Retinoblastoma from January 2017 through December 2022. Information obtained from their records included biosocioeconomic data, symptoms, lag time from initial symptoms, staging, treatment and outcome (dead or alive). Fifty-three patients, aged 6 to 88 months on first hospital presentation were recruited. There were 29(54.7%) females and 20(37.7%) patients died. Parental low socioeconomic class, rural residence and poor nutrition occurred more in those that survived, though not significantly (p>0.05). Median(interquartile) age at diagnosis [24(18-36) months, p=0.005] and lag time [13(6-20) months, p=0.274] were low in the survived group. Bilateral Retinoblastoma (20.8%,p=0.002), brain metastasis (22.6%,p<0.001), IRSS IV (18.9%,p=0.01) and relapse (34%,p<0.001) occurred more among the patients that died. The overall survival (OS) was 22(11.77-32.23) months with 1-year OS of 63%. Treatment with only chemotherapy [HR 4.76(95%CI:1.726-13.128)], incomplete chemotherapy [HR 5.61(95%CI:1.271-24.741)], relapse [HR 5.98(95%CI:1.376-25.983)] and eye surgery after 3 chemotherapy cycles [HR 8.22(95%CI:1.087-62.239)] were predictors of mortality. Early presentation of retinoblastoma especially of advanced and bilateral disease may lead to improved survival if chemotherapy and eye surgery are appropriately performed. Routine screening and immediate referral of retinoblastoma particularly in rural areas are recommended.


A new solid oral tablet formulation of posaconazole: a randomized clinical trial to investigate rising single- and multiple-dose pharmacokinetics and safety in healthy volunteers.
Posaconazole is an extended-spectrum triazole with proven efficacy as antifungal treatment and prophylaxis. The marketed oral suspension should be taken with food to maximize systemic absorption. A new solid oral tablet has been developed with improved bioavailability that can be administered without regard to food. The aim of this study was to evaluate rising single- and multiple-dose pharmacokinetics, safety and tolerability of the new tablet. This was a single-centre, randomized, placebo-controlled, Phase I, rising single- and multiple-dose study of healthy subjects aged 18-65 years who received a posaconazole tablet as 200 mg once daily, 200 mg twice daily or 400 mg once daily. The 24 subjects were studied in two cohorts of 12 subjects each (9 active and 3 placebo). After single or multiple oral dose administration of posaconazole tablets (200 and 400 mg), exposure increased in a dose-related manner. Peak posaconazole concentrations were attained at a median T(max) of 4-5 h. Mean half-life was similar for 200 and 400 mg posaconazole doses (25 and 26 h). The accumulation ratio upon multiple doses over 8 days was ∼3 for 200 and 400 mg once daily and ∼5 for 200 mg twice daily. C(avg) values exceeded 1300 ng/mL. The posaconazole oral tablet was safe and well tolerated, although mild, transient elevations in liver function were reported in some patients. Posaconazole exposure increased in a dose-related manner. The pharmacokinetics of this new solid oral tablet of posaconazole supports the clinical evaluation of once-daily dosing regimens for fungal infections.


Drug Targeting of Plasminogen Activator Inhibitor-1 Inhibits Metabolic Dysfunction and Atherosclerosis in a Murine Model of Metabolic Syndrome.
Enhanced expression of PAI-1 (plasminogen activator inhibitor-1) has been implicated in atherosclerosis formation in humans with obesity and metabolic syndrome. However, little is known about the effects of pharmacological targeting of PAI-1 on atherogenesis. This study examined the effects of pharmacological PAI-1 inhibition on atherosclerosis formation in a murine model of obesity and metabolic syndrome. Approach and Results: LDL receptor-deficient (ldlr-/-) mice were fed a Western diet high in cholesterol, fat, and sucrose to induce obesity, metabolic dysfunction, and atherosclerosis. Western diet triggered significant upregulation of PAI-1 expression compared with normal diet controls. Addition of a pharmacological PAI-1 inhibitor (either PAI-039 or MDI-2268) to Western diet significantly inhibited obesity and atherosclerosis formation for up to 24 weeks without attenuating food consumption. Pharmacological PAI-1 inhibition significantly decreased macrophage accumulation and cell senescence in atherosclerotic plaques. Recombinant PAI-1 stimulated smooth muscle cell senescence, whereas a PAI-1 mutant defective in LRP1 (LDL receptor-related protein 1) binding did not. The prosenescent effect of PAI-1 was blocked by PAI-039 and R2629, a specific anti-LRP1 antibody. PAI-039 significantly decreased visceral adipose tissue inflammation, hyperglycemia, and hepatic triglyceride content without altering plasma lipid profiles. Pharmacological targeting of PAI-1 inhibits atherosclerosis in mice with obesity and metabolic syndrome, while inhibiting macrophage accumulation and cell senescence in atherosclerotic plaques, as well as obesity-associated metabolic dysfunction. PAI-1 induces senescence of smooth muscle cells in an LRP1-dependent manner. These results help to define the role of PAI-1 in atherosclerosis formation and suggest a new plasma-lipid-independent strategy for inhibiting atherogenesis.


Properties of Alkali Activated Lightweight Aggregate Generated from Sidoarjo Volcanic Mud (Lusi), Fly Ash, and Municipal Solid Waste Incineration Bottom Ash.
Production of artificial lightweight aggregate (LWA) from industrial by-products or abundant volcanic mud is a promising solution to prevent damaging the environment due to the mining of natural aggregate. However, improvements are still needed in order to control the high water absorption of LWA and strength reduction in resulting concrete or mortar. Hence in this research, fly ash, municipal solid waste incineration bottom ash (MSWI BA), and Sidoarjo volcanic mud (Lusi) were employed as a precursor and activated using NaOH 6 M and Na2SiO3 in producing LWA. The influence of the type of the precursors on the physical properties of resulting LWA was investigated. The effect of replacing natural fine aggregate with the resulting LWA on the compressive strength and volume density of mortar was also determined. Finer particles, a high amount of amorphous phase, and low loss on ignition (LOI) of the raw material improved the properties of resulting LWA. Mortar compressive strength was decreased by 6% when replacing 16% by volume of natural fine aggregate with fly ash based LWA. Compared to the expanded clay LWA, the properties of alternative LWAs in this study were slightly, but not significantly, inferior. Alternative LWA becomes attractive when considering that expanded clay LWA requires more energy during the sintering process.


Multibeam 3D Underwater SLAM with Probabilistic Registration.
This paper describes a pose-based underwater 3D Simultaneous Localization and Mapping (SLAM) using a multibeam echosounder to produce high consistency underwater maps. The proposed algorithm compounds swath profiles of the seafloor with dead reckoning localization to build surface patches (i.e., point clouds). An Iterative Closest Point (ICP) with a probabilistic implementation is then used to register the point clouds, taking into account their uncertainties. The registration process is divided in two steps: (1) point-to-point association for coarse registration and (2) point-to-plane association for fine registration. The point clouds of the surfaces to be registered are sub-sampled in order to decrease both the computation time and also the potential of falling into local minima during the registration. In addition, a heuristic is used to decrease the complexity of the association step of the ICP from O(n2) to O(n) . The performance of the SLAM framework is tested using two real world datasets: First, a 2.5D bathymetric dataset obtained with the usual down-looking multibeam sonar configuration, and second, a full 3D underwater dataset acquired with a multibeam sonar mounted on a pan and tilt unit.


Corneal arcus and hyperlipoproteinaemia.
A corneal arcus is a lipid deposition mainly consisting of cholesterol and phospholipid. Its prevalence varies in different populations and races, increases with age and is greater in the male. Earlier studies related the presence and severity of a corneal arcus to plasma lipid levels and linked it with certain familial hyperlipidaemias. The recent reclassification of such disorders in terms of hyperlipoproteinaemia now links the premature occurrence of an arcus with familial Type II and III hyperlipoproteinaemia. Other rare plasma lipid disorders in which corneal opacities occur are Tangier Disease and lecithin:cholesterol acyl transferase deficiency.


[Immunoenzyme determination of the total level of ceruloplasmin in the serum from patients with hepatocerebral dystrophy].
An immunoenzymatic method for ceruloplasmin analysis (IEA) based on the use of horseradish peroxidase-labelled monospecific antibodies as markers has been developed. IEA can be used for direct measurements of ceruloplasmin in blood serum, as can be evidenced from the coincidence of calibration plots obtained after the use of potassium-phosphate buffer and ceruloplasmin-free sera. The procedure allows the determination of the total content of ceruloplasmin present in the blood sera of patients with hepatocerebral dystrophies both in the active and inactive forms. The minimum ceruloplasmin concentration detectable by this method is 5 x 10(-9) g/ml. The method was used to determine ceruloplasmin levels in the blood of patients with various grades of hepatocerebral dystrophy. Analysis of blood sera from 6 patients revealed that the ceruloplasmin concentrations determined by IEA were very close, whereas the oxidase activities of this protein differed more than 7-fold. The amount of enzymatically active ceruloplasmin as determined from the oxidase activity made up to 10-68% of the total ceruloplasmin content in the sera, depending of the severity of the pathology.


Hormonal contraception and cardiovascular system.
Hormonal contraception is the most widely used method to prevent unplanned pregnancies. The literature has shown an association between cardiovascular risk and use of hormone therapy. With the purpose of providing better guidelines on contraception methods for women with risk factors for cardiovascular disease, we have reviewed the literature on the subject. This review describes the latest data from the scientific literature concerning the influence of hormonal contraceptives on arterial thrombosis, venous thrombosis and systemic high blood pressure, which are diseases that have become increasingly prevalent among young females.


Second-line paclitaxel/carboplatin versus vinorelbine/carboplatin in patients who have advanced non-small-cell lung cancer pretreated with non-platinum-based chemotherapy: a multicenter randomized phase II study.
This study evaluates the activity and toxicity of the paclitaxel/carboplatin (PC) doublet versus vinorelbine/carboplatin (VC) doublet as second-line treatment in patients who have advanced non-small-cell lung cancer (NSCLC). Patients pretreated with front-line docetaxel and gemcitabine were randomized to receive either PC (n = 75), which consisted of paclitaxel at a dose of 140 mg/m(2) and carboplatin area under the curve (AUC3), or VC (n = 78), which consisted of vinorelbine at a dose of 45 mg/m(2) orally and carboplatin AUC3; both drugs were administered on days 1 and 15. The overall response rate was 18.6% (95% confidence interval, 9.85%-27.49%; one complete and 13 partial responses) in the PC arm and 7.7% (95% confidence interval, 1.78%-13.61%; one complete and five partial responses) in the VC arm (P = .056). Median time to tumor progression was 3.5 months (range, 0.3 - 23.73 months) and 3.07 months (range, 0.37-18.5) in the PC and VC arm, respectively (P = .287). Median overall survival was 7.83 months (range, 0.3-45.03 months) and 7.60 months (range, 0.5-30.27 months) for PC and VC arms, respectively (P value = .633). Chemotherapy was well-tolerated and grade III/IV toxicities were relatively infrequent. No toxic deaths were observed. Platinum-based doublets with either paclitaxel or vinorelbine in patients with advanced/metastatic NSCLC pretreated with front-line docetaxel/gemcitabine show comparable efficacy when used in the second-line setting.


Renomegaly and acute kidney injury as primary manifestations of non-Hodgkin's lymphoma: a report of three cases.
In adults with non-Hodgkin's lymphoma, renal enlargement and acute kidney injury occur infrequently at first presentation, especially in T lymphocytic lymphomas. We report three cases of non-Hodgkin's lymphoma with acute renal injury and bilateral renal enlargement. At diagnosis, one patient presented with an adrenal mass, one patient's lymph node biopsy was consistent with a renal biopsy, and one patient had primary renal lymphoma with no extrarenal disease. Assessment of renal pathology in Case 2 and Case 3 showed interstitial lymphocyte infiltration; the pathological types were non-Hodgkin's diffuse large B lymphoma originating from activated B cells outside germinal centers and non-Hodgkin's T-lymphoblastic lymphoma/leukemia, respectively. Case 1 did not receive anti-lymphoma therapy and died from infection and multiple organ failure within 1 month of hospitalization. Case 2 received eight courses of R-CHOP; her lymphoma recurred 2 years after diagnosis and she died from severe pulmonary infection 3 years after diagnosis. Case 3 received hyper-CVAD regularly and achieved stable renal function; this patient remains under follow-up. Renal lymphoma may have diverse manifestations, especially primary renal lymphoma without extrarenal involvement. Nephrologists should pay careful attention to these manifestations to ensure accurate diagnosis.


Sport-Specific Capacity to Use Elastic Energy in the Patellar and Achilles Tendons of Elite Athletes.
Introduction: During running and jumping activities, elastic energy is utilized to enhance muscle mechanical output and efficiency. However, training-induced variations in tendon spring-like properties remain under-investigated. The present work extends earlier findings on sport-specific profiles of tendon stiffness and cross-sectional area to examine whether years of distinct loading patterns are reflected by tendons' ability to store and return energy. Methods:Ultrasound scans were performed to examine the morphological features of knee extensor and plantar flexor muscle-tendon units in elite ski jumpers, distance runners, water polo players, and sedentary controls. Tendon strain energy and hysteresis were measured with combined motion capture, ultrasonography, and dynamometry. Results: Apart from the fractional muscle-to-tendon cross-sectional area ratio being lower in the knee extensors of ski jumpers (-31%) and runners (-33%) than in water polo players, no difference in the considered muscle-tendon unit morphological features was observed between groups. Similarly, no significant difference in tendon energy storage or energy return was detected between groups. In contrast, hysteresis was lower in the patellar tendon of ski jumpers (-33%) and runners (-30%) compared to controls, with a similar trend for the Achilles tendon (significant interaction effect and large effect sizes η2 = 0.2). Normalized to body mass, the recovered strain energy of the patellar tendon was ~50% higher in ski jumpers than in water polo players and controls. For the Achilles tendon, recovered strain energy was ~40% higher in ski jumpers and runners than in controls. Discussion: Advantageous mechanical properties related to tendon spring-like function are observed in elite athletes whose sport require effective utilization of elastic energy. However, the mechanisms underpinning the better tendon capacity of some athletes to retain elastic energy could not be ascribed to intrinsic or morphological features of the lower limb muscle-tendon unit.


Pharmacokinetics of biliary excretion of physostigmine in rat.
The biliary excretion of Phy and its metabolites was studied in the rat after i.v. administration of 3H-Phy (100 micrograms/kg). About 60% of the radioactivity was excreted in three hours. The unchanged drug was excreted to the extent of about 23% of the dose and the metabolites were excreted as 37% of the dose. The half-life of Phy was found to be 21.6 +/- 1.8 min (mean +/- S.E.) as determined in bile, 52.1 +/- 9.1 min as determined from serum, and the systemic clearance of Phy was .63 +/- 2.6 ml/min. The mean Kel was found to be .012 min-1. The biliary excretion rate constant was determined to be .014 +/- .001 min-1 for total radioactivity and .008 +/- .002 min-1 for Phy. Biliary clearance of Phy was .18 ml/min. The serum to bile ratio of Phy at 15-30 min was 5.11, indicating rapid concentration of Phy in the bile. Phy may stimulate bile flow in the rat, particularly at early times. The mass spectrum of extracted Phy from bile at m/e 275, 218, 175, 161 and 160 confirmed its identify. From these studies it appears that bile is a significant route of excretion of Phy in the rat.


Optical turbulence parameters characterized via optical measurements over a 2.33 km free-space laser path.
Optical turbulence research contributes to improved laser communications, adaptive optics, and long-range imaging systems. This paper presents experimental measurements of scintillation and focal spot displacement to obtain optical turbulence information along a near-horizontal 2.33 km free-space laser propagation path. Calculated values for the refractive index structure constant (C(n)(2)) and Fried parameter (r0) are compared to scintillometer-based measurements for several cases in winter and spring. Optical measurements were investigated using two different laser sources for the first and second parts of the experiment. Scintillation index estimates from recorded signal intensities were corrected to account for aperture averaging. As a result, we found that an earlier calculation algorithm based on analysis of log-amplitude intensity variance was the best estimator of optical turbulence parameters over the propagation path considered.


Targeting the glyoxalase pathway enhances TRAIL efficacy in cancer cells by downregulating the expression of antiapoptotic molecules.
Methylglyoxal is an essential component in glycolysis and is known to be an inducer of apoptosis. Glyoxalase I (GLO1) metabolizes and inactivates methylglyoxal. GLO1 is known to be overexpressed in cancer cells and causes resistance to anticancer agents. We show for the first time that methylglyoxal treatment or the silencing of GLO1 enhances sensitivity to the promising anticancer agent TRAIL in malignant tumor cells. Methylglyoxal suppressed the expression of antiapoptotic factors, X-linked inhibitor of apoptosis protein (XIAP), survivin, cIAP1, Bcl-2, and Bcl-xL, without affecting TRAIL receptors, DR4 and DR5. Knockdown of XIAP or survivin by siRNA also enhanced TRAIL-induced apoptosis, indicating that downregulation of XIAP and survivin expression by methylglyoxal contributes to the enhancement of TRAIL activity. Furthermore, methylglyoxal decreased NF-κB activity with or without TRAIL treatment. On the other hand, the knockdown of GLO1 by siRNA enhanced TRAIL-induced apoptosis via the downregulation of XIAP and survivin expression. In conclusion, our results strongly suggest that sensitivity to TRAIL is increased by inhibition of the glyoxalase pathway and that the combination of TRAIL with methylglyoxal or glyoxalase inhibitors may be useful for a novel combination chemotherapy.


Counter-regulation of interleukin-1alpha (IL-1alpha) and IL-1 receptor antagonist in murine keratinocytes.
Interleukin-1alpha (IL-1alpha) is a potent proinflammatory cytokine constitutively expressed by keratinocytes, which also synthesize a specific inhibitor of IL-1 activity, intracellular IL-1 receptor antagonist (IL-1ra). Although homeostatic regulation of the IL-1 system in keratinocytes has long been suspected, there is currently little evidence for this. To explore this issue, the PAM212 murine keratinocyte cell line was exposed to increasing concentrations of either IL-1alpha or IL-1ra and the opposing ligand was assessed by ELISA. Release of IL-1ra was induced following stimulation by murine IL-1alpha in a concentration-dependent manner and, conversely, IL-1ra stimulation increased IL-1alpha release. To determine whether a similar homeostatic circuit operates in vivo, epidermis from transgenic mice in which overexpression of IL-1alpha or IL-1ra was targeted to keratinocytes was analyzed. Epidermal sheets derived from IL-1alpha transgenic mice released eight times more IL-1ra than those from wild-type mice following ex vivo culture and similarly, IL-1alpha release was increased 3-4-fold in epidermal sheets derived from IL-1ra transgenic epidermis, Use of specific neutralizing antibodies against type I and type II IL-1 receptors indicated that the counter-regulation mechanism is mediated extracellularly through the type I IL-1 receptor alone. Taken together, these observations provide the first demonstration of mutual counter-regulation of IL-1 receptor ligands in keratinocytes.


Identification of different measles virus-specific antibodies in the serum and cerebrospinal fluid from patients with subacute sclerosing pancencephalitis and multiple sclerosis.
Matched serum and cerebrospinal fluid (CSF) samples from eight cases of subacute sclerosing panencephalitis (SSPE) and 15 cases of multiple sclerosis (MS) were characterized in neutralization, hemolysis-inhibition (HLI), hemagglutination-inhibition (HI) with Tween 80-ether-treated antigen, complement-fixation (CF), and immunodiffusion tests. CF tests were carried out with crude virus material, purified nucleocapsids, and small particle hemagglutinin as antigens. A certain diversity in the relative content of antibodies against different virus products in various sera was found. There was a high degree of correlation between titers of neutralizing and HLI antibodies, but a less strict correlation between titers of HLI and HI antibodies. Serum samples from two cases of MS and one case of SSPE contained high titers of HLI and neutralizing antibodies in the presence of only low titers of HI antibodies demonstrable with Tween 80-ether-treated antigen. The major fraction of antibodies detected in CF and immunodiffusion tests reacted with nucleocapsids. There was a tendency of nucleocapsid CF antibody titers, as compared to neutralization and HLI antibody titers, to be higher in samples from patients with SSPE than from cases of MS. No significant differences were found between antibody titers recorded in neutralization, HLI, and HI tests carried out with two different measles virus strains, Edmonston and a strain (LEC) derived from a case of SSPE. Comparison of antibodies against measles virus products and, as a reference, against a group-specific vertex capsomer antigen of adenovirus in matched serum and CSF samples revealed a production of measles virus-specific antibodies within the central nervous system of all cases of SSPE and 8 out of 15 cases of MS.


[Community-acquired urinary infection: the in vitro activity of trimethoprim and cotrimoxazole].
To evaluate the in vitro sensitivities of trimetoprim (TMP) in our area and to compare them with those to co-trimoxazole (CMX) a prospective study was carried out in females with uncomplicated lower urinary tract infection (UCLUTI), as a preliminary step for the possible use of a monodose of TMP in these patients. Fifty-five cases of UCLUTI were included. Escherichia coli was the predominating organism (70.9%). The general sensitivity to CMX was 80% and that to TMP 76.4%. The E. coli sensitivity to CMX was 79.5%, and 76.9% to TMP. The difference in the sensitivities to both antimicrobials was not statistically significant (p = 0.5). The routine introduction of TMP in the antibiogram would permit to evaluate the resistance to this antimicrobial in each area and, on the basis of the results, to assess the effectiveness of TMP in the treatment of UCLUTI.


Injury in young people with intellectual disability: descriptive epidemiology.
To assess the public health importance of injury in a representative sample of young people with intellectual disability relative to the general population. This study forms part of the Australian Child and Adolescent Development (ACAD) program examining emotional and behavioural problems in a cohort of young people with intellectual disability (IQ<70). The program has collected extensive biopsychosocial data from carers of subjects at two time intervals, 1990 (n = 579) and 1996 (n = 465). Carer report of medically attended injury to subjects was collected for the first time during 1996 (age 5-29 years) and supplemented with medical record injury data from hospitals and general practitioners for 147 of the ACAD sample and 110 supplementary subjects. These data were compared with general population injury data to assess relative epidemiological differences. Annual injury mortality and morbidity rates were 150/100,000 and 55.6/1000 persons, with age standardised mortality and morbidity ratios of 8 and 2 respectively. Males and females had similar injury rates. The rate for injury hospitalisations was twice that of the general population. Falls were more common and transport injury and intentional injury less common causes of injury morbidity compared with general population. The patterns of cause, circumstances, and severity of injury in young people with intellectual disability have more similarities with younger children than with their same age group in the general population. This study should alert clinicians and others to the increased risk for injury and possible further handicap in this population. It is essential that injury prevention programs be implemented and evaluated for their effectiveness in reducing the substantial additional burden of suffering, care and cost resulting from injury to young people with intellectual disability.


Isotopic ratio of 129I/127I in seaweed measured by neutron activation analysis with gamma-gamma coincidence.
129I is a long-lived (1.6 x 10(7) y) radionuclide that is produced in nature as the result of spontaneous fission of heavy elements and reaction of xenon with cosmic rays. Recently, however, artificial sources from nuclear power plants and nuclear test explosions have become a significant component of environmental radioactive iodine. Coincidence gamma-ray detection using Ge detectors makes it possible to simultaneously resolve the numerous gamma-rays produced by neutron activation. In this study, the coincidence gamma-ray detection technique was combined with neutron activation analysis to determine the radioactive iodine composition of seaweed. The ratio of 129I/127I in this common Japanese food item collected from the Ibaraki prefecture has been derived without the need for radiochemical purification. The isotopic ratio of 129I/127I in Kajime algae is 3.5(5) x 10(-10).


Protein turnover in the whole animal.
Methods of measuring whole body protein turnover (WBPT) are discussed briefly. In a range of species from mouse to cow WBPT in the adult bears a fairly constant relationship to metabolic mass and to basal oxygen consumption. In young animals WBPT per unit metabolic mass is higher than in adults. During growth there appears to be an increase in the rate of protein breakdown as well as of synthesis. Of all tissues which have been studied, muscle shows the greatest developmental changes in the rate of protein synthesis. In the absence of food, for example during the night, the rate of protein turnover falls. Studies on the effects of different levels of protein and energy supply have not so far given a constant picture. Very few measurements of WBPT in disease states have been made. Preliminary results suggest that the loss of nitrogen after injury may be due to a block in synthesis rather than to an increase in body protein breakdown.


The use of deconvolution techniques to identify the fundamental mixing characteristics of urban drainage structures.
Mixing and dispersion processes affect the timing and concentration of contaminants transported within urban drainage systems. Hence, methods of characterising the mixing effects of specific hydraulic structures are of interest to drainage network modellers. Previous research, focusing on surcharged manholes, utilised the first-order Advection-Dispersion Equation (ADE) and Aggregated Dead Zone (ADZ) models to characterise dispersion. However, although systematic variations in travel time as a function of discharge and surcharge depth have been identified, the first order ADE and ADZ models do not provide particularly good fits to observed manhole data, which means that the derived parameter values are not independent of the upstream temporal concentration profile. An alternative, more robust, approach utilises the system's Cumulative Residence Time Distribution (CRTD), and the solute transport characteristics of a surcharged manhole have been shown to be characterised by just two dimensionless CRTDs, one for pre- and the other for post-threshold surcharge depths. Although CRTDs corresponding to instantaneous upstream injections can easily be generated using Computational Fluid Dynamics (CFD) models, the identification of CRTD characteristics from non-instantaneous and noisy laboratory data sets has been hampered by practical difficulties. This paper shows how a deconvolution approach derived from systems theory may be applied to identify the CRTDs associated with urban drainage structures.


Cytotoxic CD4+ tissue-resident memory T cells are associated with asthma severity.
Patients with severe uncontrolled asthma represent a distinct endotype with persistent airway inflammation and remodeling that is refractory to corticosteroid treatment. CD4+ TH2 cells play a central role in orchestrating asthma pathogenesis, and biologic therapies targeting their cytokine pathways have had promising outcomes. However, not all patients respond well to such treatment, and their effects are not always durable nor reverse airway remodeling. This observation raises the possibility that other CD4+ T cell subsets and their effector molecules may drive airway inflammation and remodeling. We performed single-cell transcriptome analysis of >50,000 airway CD4+ T cells isolated from bronchoalveolar lavage samples from 30 patients with mild and severe asthma. We observed striking heterogeneity in the nature of CD4+ T cells present in asthmatics' airways, with tissue-resident memory T (TRM) cells making a dominant contribution. Notably, in severe asthmatics, a subset of CD4+ TRM cells (CD103-expressing) was significantly increased, comprising nearly 65% of all CD4+ T cells in the airways of male patients with severe asthma when compared to mild asthma (13%). This subset was enriched for transcripts linked to T cell receptor activation (HLA-DRB1, HLA-DPA1) and cytotoxicity (GZMB, GZMA) and, following stimulation, expressed high levels of transcripts encoding for pro-inflammatory non-TH2 cytokines (CCL3, CCL4, CCL5, TNF, LIGHT) that could fuel persistent airway inflammation and remodeling. Our findings indicate the need to look beyond the traditional T2 model of severe asthma to better understand the heterogeneity of this disease. This research was funded by the NIH.


Comparing pairwise-additive and many-body generalized Born models for acid/base calculations and protein design.
Generalized Born (GB) solvent models are common in acid/base calculations and protein design. With GB, the interaction between a pair of solute atoms depends on the shape of the protein/solvent boundary and, therefore, the positions of all solute atoms, so that GB is a many-body potential. For compute-intensive applications, the model is often simplified further, by introducing a mean, native-like protein/solvent boundary, which removes the many-body property. We investigate a method for both acid/base calculations and protein design that uses Monte Carlo simulations in which side chains can explore rotamers, bind/release protons, or mutate. The fluctuating protein/solvent dielectric boundary is treated in a way that is numerically exact (within the GB framework), in contrast to a mean boundary. Its originality is that it captures the many-body character while retaining the residue-pairwise complexity given by a fixed boundary. The method is implemented in the Proteus protein design software. It yields a slight but systematic improvement for acid/base constants in nine proteins and a significant improvement for the computational design of three PDZ domains. It eliminates a source of model uncertainty, which will facilitate the analysis of other model limitations. © 2017 Wiley Periodicals, Inc.


Evaluation of trap types and food attractants for Rhagoletis cerasi (Diptera: Tephritidae).
Trapping experiments were conducted during the period of flight activity of the cherry fruit fly Rhagoletis cerasi L. in the area of Thessaloniki, northern Greece, during the years 1993-1997 to test several traps alone and in combination with different food attractants. Yellow sticky-coated visual traps were more effective than McPhail-type traps baited with different food attractants. Of the visual traps, the most effective was the yellow Rebell trap. The Rebell trap, provided with a dispenser containing a slow release formulation of ammonium acetate attached to the lower part of the trap, was found to be the most effective of all treatments tested, capturing approximately 50% more R. cerasi flies than the Rebell trap without any attractant. Ammonium carbonate dispensers did not increase the performance of Rebell traps. More mature females were captured in Rebell traps baited with or without ammonium acetate than in McPhail-type traps baited with ammonium acetate. This study demonstrates that Rebell traps baited with an ammonium acetate dispenser can provide a more effective tool for monitoring and mass trapping of R. cerasi than the currently used unbaited Rebell traps.


Industrial scale-up of pH-controlled liquid hot water pretreatment of corn fiber for fuel ethanol production.
The pretreatment of cellulose in corn fiber by liquid hot water at 160 degrees C and a pH above 4.0 dissolved 50% of the fiber in 20 min. The pretreatment also enabled the subsequent complete enzymatic hydrolysis of the remaining polysaccharides to monosaccharides. The carbohydrates dissolved by the pretreatment were 80% soluble oligosaccharides and 20% monosaccharides with <1% of the carbohydrates lost to degradation products. Only a minimal amount of protein was dissolved, thus enriching the protein content of the undissolved material. Replication of laboratory results in an industrial trial at 43 gallons per minute (163 L/min) of fiber slurry with a residence time of 20 min illustrates the utility and practicality of this approach for pretreating corn fiber. The added costs owing to pretreatment, fiber, and hydrolysis are equivalent to less than 0.84 dollars/gal of ethanol produced from the fiber. Minimizing monosaccharide formation during pretreatment minimized the formation of degradation products; hence, the resulting sugars were readily fermentable to ethanol by the recombinant hexose and by pentose-fermenting Saccharomyces cerevisiae 424A(LNH-ST) and ethanologenic Escherichia coli at yields >90% of theoretical based on the starting fiber. This cooperative effort and first successful trial opens the door for examining the robustness of the pretreatment system under extended run conditions as well as pretreatment of other cellulose-containing materials using water at controlled pH.


Inhibition of the Ubc9 E2 SUMO-conjugating enzyme-CRMP2 interaction decreases NaV1.7 currents and reverses experimental neuropathic pain.
We previously reported that destruction of the small ubiquitin-like modifier (SUMO) modification site in the axonal collapsin response mediator protein 2 (CRMP2) was sufficient to selectively decrease trafficking of the voltage-gated sodium channel NaV1.7 and reverse neuropathic pain. Here, we further interrogate the biophysical nature of the interaction between CRMP2 and the SUMOylation machinery, and test the hypothesis that a rationally designed CRMP2 SUMOylation motif (CSM) peptide can interrupt E2 SUMO-conjugating enzyme Ubc9-dependent modification of CRMP2 leading to a similar suppression of NaV1.7 currents. Microscale thermophoresis and amplified luminescent proximity homogeneous alpha assay revealed a low micromolar binding affinity between CRMP2 and Ubc9. A heptamer peptide harboring CRMP2's SUMO motif, also bound with similar affinity to Ubc9, disrupted the CRMP2-Ubc9 interaction in a concentration-dependent manner. Importantly, incubation of a tat-conjugated cell-penetrating peptide (t-CSM) decreased sodium currents, predominantly NaV1.7, in a model neuronal cell line. Dialysis of t-CSM peptide reduced CRMP2 SUMOylation and blocked surface trafficking of NaV1.7 in rat sensory neurons. Fluorescence dye-based imaging in rat sensory neurons demonstrated inhibition of sodium influx in the presence of t-CSM peptide; by contrast, calcium influx was unaffected. Finally, t-CSM effectively reversed persistent mechanical and thermal hypersensitivity induced by a spinal nerve injury, a model of neuropathic pain. Structural modeling has now identified a pocket-harboring CRMP2's SUMOylation motif that, when targeted through computational screening of ligands/molecules, is expected to identify small molecules that will biochemically and functionally target CRMP2's SUMOylation to reduce NaV1.7 currents and reverse neuropathic pain.