Safety of early postoperative hydrotherapy in dogs undergoing thoracolumbar hemilaminectomy.
Hydrotherapy is a common part of the postoperative care after surgical treatment of thoracolumbar intervertebral disc extrusion in dogs. There are currently no guidelines on when to commence hydrotherapy after surgery. Early hydrotherapy may be associated with an increased risk of postoperative complications including surgical site infection and neurological deterioration. The goal of this study was to report the prevalence and types of postoperative complications in dogs receiving early hydrotherapy. Eighty-three dogs commencing hydrotherapy within 5 days after surgical treatment for thoracolumbar intervertebral disc extrusion were included in a retrospective descriptive study. All postoperative complications were recorded. Ten minor and 16 major complications were recorded in a total of 26 dogs. The majority of recorded complications were unlikely, but not excluded to be, caused by the initiation time of hydrotherapy. One dog developed a surgical site infection and one dog had a confirmed further extrusion of the originally operated intervertebral disc, which may have been influenced by early initiation of hydrotherapy. Early hydrotherapy is possibly associated with occurrence of postoperative complications. Associations between the reported complications and early hydrotherapy, and possible benefits of early hydrotherapy, need to be further investigated before early initiation of hydrotherapy can be recommended.


Effects of various knee-raising postures in a supine position on swelling in the lower legs and feet in elderly women.
The present study examined the effects of different knee-raising postures in the supine position on swelling in the lower legs and feet of elderly women. Seven elderly women with no illness-related swelling maintained a supine position for 40 min on a bed under four knee-raising postures: downhill, raising the knees and further raising the feet; horizon, raising the knees with the lower legs in a horizontal position; mountain, raising the knees to a height above the feet; and control, stretching the knees without elevation of the feet. While maintaining a supine position, downhill and horizon conditions showed significant decreases in circumferences and increased bio-impedance at all measured positions of the calf and foot, compared to the control condition. In contrast, the mountain condition did not show decreases in all circumferences. No significant differences were found for heart rate, blood pressure or scores of subjective comfort in any body areas among the conditions. These results suggest that the downhill and horizon conditions have effects on lower leg swelling without causing additional discomfort or circulatory strain.


TU-100 (Daikenchuto) and ginger ameliorate anti-CD3 antibody induced T cell-mediated murine enteritis: microbe-independent effects involving Akt and NF-κB suppression.
The Japanese traditional medicine daikenchuto (TU-100) has anti-inflammatory activities, but the mechanisms remain incompletely understood. TU-100 includes ginger, ginseng, and Japanese pepper, each component possessing bioactive properties. The effects of TU-100 and individual components were investigated in a model of intestinal T lymphocyte activation using anti-CD3 antibody. To determine contribution of intestinal bacteria, specific pathogen free (SPF) and germ free (GF) mice were used. TU-100 or its components were delivered by diet or by gavage. Anti-CD3 antibody increased jejunal accumulation of fluid, increased TNFα, and induced intestinal epithelial apoptosis in both SPF and GF mice, which was blocked by either TU-100 or ginger, but not by ginseng or Japanese pepper. TU-100 and ginger also blocked anti-CD3-stimulated Akt and NF-κB activation. A co-culture system of colonic Caco2BBE and Jurkat-1 cells was used to examine T-lymphocyte/epithelial cells interactions. Jurkat-1 cells were stimulated with anti-CD3 to produce TNFα that activates epithelial cell NF-κB. TU-100 and ginger blocked anti-CD3 antibody activation of Akt in Jurkat cells, decreasing their TNFα production. Additionally, TU-100 and ginger alone blocked direct TNFα stimulation of Caco2BBE cells and decreased activation of caspase-3 and polyADP ribose. The present studies demonstrate a new anti-inflammatory action of TU-100 that is microbe-independent and due to its ginger component.


The use of monoclonal antibodies for treatment of autoimmune disease.
Over the past decade monoclonal antibodies have been successfully employed in a number of animal models of autoimmune disease. We have used antibodies to the class II gene products of the major histocompatibility complex, the CD4 molecule on helper T cells, and the T-cell receptor. Monoclonal anti-class II antibodies have been administered to treat paralytic disease in the animal model of multiple sclerosis--experimental allergic encephalomyelitis. These antibodies not only reverse acute paralytic disease but also decrease the number of relapses in a model of relapsing/remitting multiple sclerosis when given after the first attack. The advantage of this form of therapy is that it is haplotype specific. In other words, in a heterozygous individual it is possible to block the major histocompatibility gene associated with disease susceptibility while leaving other major histocompatibility gene products free for antigen presentation. Thus, animals given this form of immunotherapy are not significantly immunosuppressed. Antibodies to the CD4 molecule have been equally effective in treating animal models of autoimmunity. We and others have reversed ongoing paralysis in experimental autoimmune encephalomyelitis. Relapses have been diminished after the administration of anti-CD4. Antibodies to CD4 have been used successfully to treat animal models of systemic lupus erythematosus, rheumatoid arthritis and myasthenia gravis. Recent trials with anti-CD4 have been successful in the treatment of rheumatoid arthritis and cutaneous T-cell lymphoma. The latter trial employed a chimeric human/mouse antibody. Antibodies to the variable region of the T-cell receptor have been employed to treat experimental autoimmune encephalomyelitis.(ABSTRACT TRUNCATED AT 250 WORDS)


Fecal shedding of Salmonella spp. by dairy cows on farm and at cull cow markets.
As part of a national study of the U.S. dairy cow population, fecal samples were collected from representative cows on 91 dairies and 97 cull dairy cow markets in 19 states. Salmonella spp. were recovered from 5.4% of milk cows, 18.1% of milk cows expected to be culled within 7 days, and 14.9% of culled dairy cows at markets. On a premise basis, Salmonella shedding in milk cows was detected on 21.1% of dairies and 66% of cull dairy cow markets. The percentage of herds with at least one cow with detectable Salmonella fecal shedding was higher during the sampling period from May through July, in herds with at least 100 milk cows, and in herds in the South region. The most common Salmonella serogroups isolated were E (30.8% of isolates) and C1 (28.6%); the most common serotypes isolated were Salmonella Montevideo (21.5% of isolates), Salmonella Cerro (13.3%), and Salmonella Kentucky (8.5%). Fecal shedding of Salmonella Typhimurium or Salmonella Typhimurium var. copenhagen was infrequent (2.8% of isolates). Most isolates (88.9%) were susceptible to all 17 antimicrobials evaluated; multiple resistance was an infrequent occurrence. This study provides information describing the distribution of Salmonella fecal shedding from dairy cows on farm and at markets and will serve as a baseline for future studies.


Enhanced phagocytosis of CD47-deficient red blood cells by splenic macrophages requires SHPS-1.
The interaction of CD47 on red blood cells (RBCs) with SHPS-1 on macrophages is implicated to prevent the phagocytosis of the former cells by the latter cells. Indeed, the rate of clearance of transfused CD47-deficient (CD47(-/-)) RBCs from the bloodstream of wild-type mice was markedly increased compared with wild-type RBCs. Conversely, the rate of clearance of transfused wild-type RBCs was markedly increased in mice that expressed a mutant form of SHPS-1 lacking most of the cytoplasmic region of the protein. However, we here found that the clearance of CD47(-/-) RBCs in SHPS-1 mutant mice was minimal. In addition, the phagocytosis of CD47(-/-) RBCs by splenic macrophages from SHPS-1 mutant mice was markedly reduced compared with wild-type macrophages. These results thus suggest an additional role for CD47 on RBCs in the negative regulation of phagocytosis by macrophages and in determination of the life span of circulating RBCs.


Physical properties of short-stretch compression bandages used to treat lymphedema.
This study examined the physical properties of six common brands of short-stretch compression bandages used to treat lymphedema. The physical properties examined were (a) maintenance of pressure over a 12-hr period, (b) variability of pressure across the width of the bandages, and (c) variability of pressure when the bandages were wrapped with a 50% overlap. The results of the study indicate that all six brands of bandages tested maintain pressure well over a 12-hr period. Each has a variance of pressure between the middle and edge of the bandage, with the edges measuring (in mmHg) between 6% and 28% lower than the middle. When the bandages were wrapped with an 50% overlap, all six brands measured fairly consistently in pressure readings (in mmHg) across the width. These results indicate that the six brands of short-stretch compression bandages tested have similar physical characteristics.


Development of measures of polyneuropathy impairment in hATTR amyloidosis: From NIS to mNIS + 7.
Hereditary transthyretin-mediated amyloidosis (hATTR amyloidosis) is a rare, life-threatening disease, caused by point mutations in the transthyretin gene. It is a heterogeneous, multisystem disease with rapidly progressing polyneuropathy (including sensory, motor, and autonomic impairments) and cardiac dysfunction. Measures used to assess polyneuropathy in other diseases have been tested as endpoints in hATTR amyloidosis clinical trials (i.e. Neuropathy Impairment Score [NIS], NIS-lower limb, and NIS + 7), yet the unique nature of the polyneuropathy in this disease has necessitated modifications to these scales. In particular, the heterogeneous impairment and the aggressive disease course have been key drivers in developing scales that better capture the disease burden and progression of polyneuropathy in hATTR amyloidosis. The modified NIS + 7 (mNIS + 7) scale was specifically designed to assess polyneuropathy impairment in patients with hATTR amyloidosis, and has been the primary endpoint in two recent, phase III studies in this disease. The mNIS + 7 uses highly standardized, quantitative, and referenced assessments to quantify decreased muscle weakness, muscle stretch reflexes, sensory loss, and autonomic impairment. Physicians using this scale in clinical trials should be specifically trained and monitored to minimize variability. This article discusses the different scales that have been/are being used to assess polyneuropathy in patients with hATTR amyloidosis, their correlation with other disease assessments, and reflects on how and why scales have evolved to the latest iteration of mNIS + 7.


External Validation of Randomized Trial Outcomes Following Carotid Interventions in the Modern Era.
An essential prerequisite of randomized controlled trials (RCTs) is "external validation," that is, results obtained in the "real world" closely replicate those from RCTs. In this respect, contemporary administrative data set registries reporting outcomes after carotid endarterectomy and carotid artery stenting (CAS) have recently reported death/stroke rates that exceed the accepted risk thresholds for intervening. The current article evaluates this controversy and offers reasons why this might continue to happen, namely: (1) difference in interventionists' competence/experience, (2) patient selection, and (3) advances in CAS technology/technique. As CAS is a continuously evolving technique, the results obtained from patients recruited into the landmark RCTs (as early as the late 1990s) do not reflect contemporary practice. Although RCTs are not always the perfect solution, the process of randomization ensures minimization of selection bias. A possible way forward may be the introduction of prospective, randomized, controlled clinical registries.


Impact of Hunger, Satiety, and Oral Glucose on the Association Between Insulin and Resting-State Human Brain Activity.
To study the interplay of metabolic state (hungry vs. satiated) and glucose administration (including hormonal modulation) on brain function, resting-state functional magnetic resonance imaging (rs-fMRI) and blood samples were obtained in 24 healthy normal-weight men in a repeated measurement design. Participants were measured twice: once after a 36 h fast (except water) and once under satiation (three meals/day for 36 h). During each session, rs-fMRI and hormone concentrations were recorded before and after a 75 g oral dose of glucose. We calculated the amplitude map from blood-oxygen-level-dependent (BOLD) signals by using the fractional amplitude of low-frequency fluctuation (fALFF) approach for each volunteer per condition. Using multiple linear regression analysis (MLRA) the interdependence of brain activity, plasma insulin and blood glucose was investigated. We observed a modulatory impact of fasting state on intrinsic brain activity in the posterior cingulate cortex (PCC). Strikingly, differences in plasma insulin levels between hunger and satiety states after glucose administration at the time of the scan were negatively related to brain activity in the posterior insula and superior frontal gyrus (SFG), while plasma glucose levels were positively associated with activity changes in the fusiform gyrus. Furthermore, we could show that changes in plasma insulin enhanced the connectivity between the posterior insula and SFG. Our results indicate that hormonal signals like insulin alleviate an acute hemostatic energy deficit by modifying the homeostatic and frontal circuitry of the human brain.


A Systematic Review and Network Meta-analysis of Novel Androgen Receptor Inhibitors in Non-metastatic Castration-resistant Prostate Cancer.
Among men with high-risk non-metastatic castrate-resistant prostate cancer (nmCRPC), we used network meta-analysis to compare non-steroidal anti-androgens (NSAAs) and stratified class-level meta-analysis to identify subgroups with particular benefit from NSAAs with androgen deprivation therapy versus androgen deprivation therapy alone. We performed a systematic review of phase III parallel-group randomized controlled trials in adult men with nmCRPC. Primary outcome was metastasis-free survival (MFS). Secondary outcomes included overall survival (OS), prostate-specific antigen (PSA) progression-free survival (PFS), and rates of grade 3 to 4 adverse events (AEs). We assessed class-level effects using random effects models; effect modification owing to subgroup effects using random-effects models to pool study-level differences; and comparative outcomes between agents using fixed-effect network models in a Bayesian framework. Three randomized controlled trials were identified. Pooled MFS, PSA-PFS, and OS were significantly greater with NSAA versus placebo (hazard ratio [HR], 0.32; 95% confidence interval [CI], 0.25-0.41; HR, 0.08; 95% CI, 0.05-0.13; and HR, 0.74; 95% CI, 0.61-0.90, respectively). Subgroup analysis demonstrated a greater benefit with NSAAs in men with Eastern Cooperative Oncology Group performance status 0 (HR, 0.30; 95% CI, 0.24-0.38) versus 1 (HR, 0.45; 95% CI, 0.36-0.56; P = .005), but no difference owing to PSA doubling time (P = .43) or use of osteoclast targeting therapy (P = .77). Bayesian analysis showed apalutamide and enzalutamide had a 56% and 44% likelihood of maximizing MFS, respectively, with subgroup analysis demonstrating these agents were preferred regardless of PSA doubling time and performance status. There was a 44%, 41%, and 15% likelihood that apalutamide, darolutamide and enzalutamide offered the greatest OS benefit, respectively. Grade 3 to 4 AEs were more common with NSAAs (odds ratio [OR], 1.47; 95% CI, 1.27-1.71) and there was a 61% chance that darolutamide was preferred. NSAAs improve survival in high-risk nmCRPC. Apalutamide and enzalutamide may result in improved oncologic outcomes. Darolutamide may result in fewer AEs. Phase IV data are needed to validate these findings.


Milk intake and carbon dioxide production of piglets determined with the doubly labelled water technique.
The present study was undertaken to study different methodological aspects of quantifying CO2 production and milk intake of suckling piglets using the doubly labelled water (DLW) technique. In total, 37 piglets were enriched intraperitoneally with DLW to study equilibration time of 18O (n = 3), to validate the estimation of milk intake and CO2 production (n = 10) of piglets fed milk replacer and to quantify milk intake and CO2 production of piglets nursed ordinarily by sows (n = 24). Enrichment of 18O in expired air was analysed without any sample preparation, whereas enrichment of 18O in serum was analysed after a minimum step of sample preparation, which included pipetting of the sample, blowing gaseous CO2 into the vial for 3 s and equilibrating for 24 h. The 18O enrichment of CO2 in expired air was constant within 30-40 min of intraperitoneal injection, suggesting that DLW was equilibrated within the body water by that time. For piglets fed milk replacer, the estimation of the daily CO2 production by the DLW method (64.0 ± 2.7 l CO2/day) was in agreement with that obtained by respiration trials (64.7 ± 1.8 l CO2/day). Furthermore, the intake of milk replacer (891 ± 63 g/day) determined by deuterium oxide (D2O) dilution was similar in magnitude to that found by weighing the milk disappearance (910 ± 58 g/day). The milk intake of piglets fed milk replacer was comparable with that of sucking piglets, but sucking piglets had a remarkably higher CO2 production than artificially reared piglets, which likely was caused by a higher intake of milk solids and a higher activity level. For sucking piglets, the daily CO2 production increased curvilinearly with increasing live weight (LW) in kg: piglet CO2 production (l/day) = 25.75 × LW - 1.01 × LW2. In conclusion, 18O equilibrates fast within the body water pool when administered intraperitoneally, and the accuracy of assessing milk intake and rate of CO2 production using the DLW technique is promising. Assessment of excess enrichment of 18O in serum proved to be robust. Finally, the CO2 production of piglets fed milk replacer differs considerably from that of sucking piglets.


Sub-Saharan Africa Tackles COVID-19: Challenges and Opportunities.
As of May 2020, the global COVID-19 pandemic had reached 187 countries with more than 3.7 million confirmed cases and 263,000 deaths. While sub-Saharan Africa (SSA) has not been spared, the extent of disease is currently far less than in Europe or North America leading some to posit that climatic, genetic or other conditions will self-limit disease in this location. Nonetheless, infections in tropical Africa continue to rise at an alarming pace with the potential to soon exceed health resource availability and to exhaust a health care workforce that is already grossly under supported and ill-equipped. This perspective outlines the context of COVID-19 disease in Africa with a focus on the distinctive challenges faced by African nations and a potential best path forward.


Agents for gallstone dissolution.
Numerous methods are presently available for gallstone dissolution, including oral bile salts; cholesterol solvents such as mono-octanoin and methyl tert-butyl ether; calcium or pigment solvents such as EDTA and polysorbate; mechanical extraction techniques through a T-tube tract or after endoscopic sphincterotomy; or fragmentation methods such as ultrasonography or electrohydraulic lithotripsy, lasers, and extracorporeal shock waves. Which, if any, of these methods will be appropriate for an individual patient depends on the type of stones, whether they are in the gallbladder or bile ducts, whether access to the biliary tree is available, the patient's age and general medical condition, and the availability of expert radiologists, endoscopists, and newer equipment. In the United States, the only available oral bile salt for cholesterol gallstone dissolution is chenodeoxycholate. Ursodeoxycholate, which is more rapid and less toxic, has not been approved by the Federal Drug Administration. These agents are most effective in thin women with small, floating, radiolucent cholesterol gallstones in a functioning gallbladder. Only about half of this small subset of patients, however, will experience partial or complete dissolution of stones in 6 to 12 months. Moreover, recurrence is very likely, and the potential toxicity of long-term therapy is unknown. Thus, for most patients, cholecystectomy remains the most cost-effective and, perhaps, safest option. Intragallbladder instillation of methyl tert-butyl ether and extracorporeal shock wave therapy are also likely to be applicable to only small subsets of patients and to be associated with high recurrence rates. In patients with retained ductal cholesterol stones and access to the biliary tree, mono-octanoin therapy is advantageous in that it can be begun as soon as cholangiography demonstrates no extravasation. In properly selected patients, a 90 percent success rate with mono-octanoin infusion can be expected within a week. Radiologic or endoscopic extraction techniques require maturation of a relatively straight T-tube tract but are not dependent on the type of stone. In the hands of experts, these techniques are highly successful. In postcholecystectomy patients without access to the biliary tree, endoscopic sphincterotomy has become the preferred method of management and can be expected to succeed in more than 90 percent of patients. At this point, the exact role for ultrasonic or electrohydraulic lithotripsy and lasers is unknown. However, these techniques may be applicable in the future in patients with retained bile duct stones in whom extraction and infusion techniques have failed.


Host immune response to tuberculous meningitis.
Tuberculous meningitis (TBM) is a severe complication of tuberculosis predominantly affecting young children. Early treatment is vital to prevent morbidity and mortality, emphasizing the importance of early diagnosis. The lack of sensitive methods for early diagnosis is the most common cause of delay. Attempts have been made to develop simplified tests for tuberculosis, but their diagnostic power remains poor. The clinical picture of TBM is mainly driven by the host's immune response to Mycobacterium tuberculosis; therefore, identification of disease-specific biomarkers may have diagnostic and therapeutic value and improve our understanding of its pathogenesis. We investigated disease-specific biomarkers of childhood TBM in a cohort of children aged 3 months-13 years with symptoms and signs suggestive of meningitis. Cerebrospinal fluid (CSF) and serum from 56 patients with and 55 patients without TBM were assessed for 28 soluble mediators. Unsupervised hierarchical clustering analysis revealed a disease-specific pattern of biomarkers for TBM relative to other types of meningitis. A biomarker-based diagnostic prediction model for childhood TBM based on CSF concentrations of interleukin 13 (cutoff value, 37.26 pg/mL), vascular endothelial growth factor (cutoff value, 42.92 pg/mL), and cathelicidin LL-37 (cutoff value, 3221.01 pg/mL) is presented with a sensitivity of 0.52 and a specificity of 0.95. These data highlight the potential of biosignatures in the host's CSF for diagnostic applications and for improving our understanding of the pathogenesis of TBM to discover strategies to prevent immunopathological sequelae.


Age differences in care practices and outcomes for hospitalized patients with cancer.
To identify age group differences in care practices and outcomes for seriously ill hospitalized patients with malignancy. Prospective cohort study (SUPPORT project). Five United States teaching hospitals; data was gathered between 1989 and 1994. Nine hundred twenty five older (age > or = 65 years), 983 middle aged (age = 45-64 years), and 274 younger (age = 18-44 years) hospitalized patients receiving care for non-small cell lung cancer, colon cancer metastasized to the liver, or multi-organ system failure associated with malignancy. Care practices and patient outcomes were determined from hospital records. Length of survival was identified using the National Death Index. After adjusting for important variables, including severity of illness (i.e., SUPPORT model estimate for 2-month survival, cancer condition), hospital site, selection to intervention and sociodemographic variables, age group differences in care practices and outcomes were identified using general linear models. Older patients with cancer had lower resource utilization during hospitalization (P < .04) and were less likely to receive cancer-related treatments (i.e., chemotherapy, platelet infusions, scheduled intravenous medications) than middle-aged and young-adult patients in the first week of hospitalization (P < or = .01). More care topics were discussed with older patients and their families then with younger patients and their families (P < .001). Length of stay and total hospital costs were lower for older and middle-aged patients than for younger patients. Although more older patients had discussions about transfer to hospice (P < .001), older patients were no more likely to be discharged with supportive care (inpatient hospice or home with home/ hospice care). Older patients died sooner than middle-aged patients (P < .01). Patient age influenced care decisions and outcomes. Older patients (age > or = 65 years) received less aggressive care, had more discussions about care decisions, and died sooner than younger patients with cancer. Younger patients had longer stays, higher hospital costs, and greater probability of rehospitalization. Although well over half of patients died within 6 months of hospitalization, few patients in any age group were discharged with supportive care. Future studies should examine age differences in palliation, as well as acute care of cancer patients across inpatient and ambulatory care settings and should assess quality of care at the end of life.


Hypermentalizing in adolescent inpatients: treatment effects and association with borderline traits.
Sharp et al. (2011) recently demonstrated that in adolescents with borderline traits the loss of mentalization is more apparent in the emergence of unusual alternative strategies (excessive theory of mind or hypermentalizing) than in the loss of the capacity per se (no mentalizing or undermentalizing). This suggests that hypermentalizing could be a worthwhile social-cognitive treatment target in adolescents with borderline traits. The aim of the current study was to examine (1) whether a reduction in excessive theory of mind or hypermentalizing is achieved between admission and discharge for adolescent inpatients; (2) whether the hypothesized reduction is more apparent in adolescents meeting criteria for BPD compared with psychiatric controls; and (3) whether other forms of mentalizing would also be sensitive to and malleable by inpatient treatment in the same way we expected hypermentalizing to be. The "Movie for the Assessment of Social Cognition" Task (Dziobek et al., 2006) was administered to consecutive admissions to an adolescent inpatient setting (n = 164) at admission and discharge, alongside measures of borderline symptomology and interview-based diagnosis of BPD. Results demonstrated that 41% (n = 68) of the sample met full or intermediate criteria for BPD on an interview-based measure of BPD. A relation between borderline traits and hypermentalizing that appears to be independent of internalizing and externalizing problems was demonstrated. Hypermentalizing, but not other forms of social-cognitive reasoning (as measured by the Child Eyes Test, Basic Empathy Scale and the Mentalizing Stories Test for Adolescents), was found to be malleable through a milieu-based inpatient treatment. Clinical implications of the findings for the organization of treatment settings for adolescents are discussed.


Emergency department at the cutting edge.
This paper reviews the experience of penetrating chest trauma over a 3-year period in one UK emergency department. A retrospective review was performed of patients assessed in the emergency department resuscitation room between 1 January 2002 and 31 December 2005. Patients with penetrating chest trauma, either isolated or in combination with other injuries, were included. A Medline search was performed using the terms 'chest', 'trauma' and 'penetrating'. A total of 120 patients presented with penetrating chest trauma. Ninety-two percent were male. Ninety-six percent (115) of the patients survived to hospital discharge. Seventy-eight percent of the patients presented at night (20.00 and 8.00 h). A single wound accounted for 52% (63) of patients, multiple wounds 43% (52) with 2% (two) gun-shot wounds and 3% (three) impalings. The mean prehospital time of patients in cardiac arrest was 42 min with a mean on-scene time of 24 min. The mean prehospital time for patients undergoing formal emergency surgery was 39 min with a mean on-scene time of 16 min. Twenty-three patients required one or more tube thoracostomies to be performed in the emergency department and six underwent emergency department thoracotomy. Sixteen patients required immediate formal emergency surgery for haemorrhage control. Penetrating chest trauma contributes significantly to our trauma workload with a high proportion of patients sustaining life-threatening injuries requiring immediate intervention. Significant prehospital delays occur. Overall mortality of 4.2% is comparable with that of a major American case series. Further education and protocol development is required to ensure that prehospital and emergency department management of these patients reflects the latest evidence-based guidelines.


High cerebrospinal fluid antioxidants and interleukin 8 are protective of hypoxic brain damage in newborns.
The objective was to explain the discrepancy in the development of hypoxic ischemic brain injury (HIE) in some asphyxiated newborns rather than others. Forty newborns were classified according to their cerebrospinal neuron-specific-enolase (CSF-NSE) levels on their 5th-day of life; group 1 with low-NSE (n = 25). The remaining 15 newborns had high-NSE and were further divided into a group with no HIE (n = 10, group 2) and another with HIE (n = 5, group 3). CSF-NSE, total-hydroperoxide (TH), biological-antioxidant-potentials (BAPs), 12 cytokines and erythropoietin (EPO) were measured. The TH/BAP gave the oxidative-stress-index (OSI). The BAPs of serial dilutions of three types of EPO were tested. CSF-NSE and TH and mean OSIs were higher in group 3. IL-8 and mean BAPs were higher in group 2 than in group 1. EPO was less detected in group 3. Serial EPO dilutions correlated with their BAPs. Compensatory antioxidants and IL-8 elevation could be protective of perinatal asphyxic brain injury. Antioxidative effect of EPO could be neuroprotective.


Vitamin D and calcium intakes from food or supplements and mammographic breast density.
A better understanding of factors that affect breast density, one of the strongest breast cancer risk indicators, may provide important clues about breast cancer etiology and prevention. This study evaluates the association of vitamin D and calcium, from food and/or supplements, to breast density in premenopausal and postmenopausal women separately. A total of 777 premenopausal and 783 post-menopausal women recruited at two radiology clinics in Quebec City, Canada, in 2001 to 2002, completed a food frequency questionnaire to assess vitamin D and calcium. Breast density from screening mammograms was assessed using a computer-assisted method. Associations between vitamin D or calcium and breast density were evaluated using linear regression models. Adjusted means in breast density were assessed according to the combined daily intakes of the two nutrients using generalized linear models. In premenopausal women, total intakes of vitamin D and calcium were inversely related to breast density (beta = -1.4; P = 0.004 for vitamin D; beta = -0.8; P = 0.0004 for calcium). In multivariate linear regression, simultaneous increments in daily total intakes of 400 IU vitamin D and 1,000 mg calcium were associated with an 8.5% (95% confidence interval, 1.8-15.1) lower mean breast density. The negative association between dietary vitamin D intake and breast density tended to be stronger at higher levels of calcium intake and vice versa. Among postmenopausal women, intakes of vitamin D and calcium were not associated with breast density. These findings show that higher intakes of vitamin D and calcium from food and supplements are related to lower levels of breast density among premenopausal women. They suggest that increasing intakes of vitamin D and calcium may represent a safe and inexpensive strategy for breast cancer prevention.


Regulation of hypoxia-inducible mRNAs by the von Hippel-Lindau tumor suppressor protein requires binding to complexes containing elongins B/C and Cul2.
The von Hippel-Lindau tumor suppressor protein (pVHL) binds to elongins B and C and posttranscriptionally regulates the accumulation of hypoxia-inducible mRNAs under normoxic (21% O2) conditions. Here we report that pVHL binds, via elongin C, to the human homolog of the Caenorhabditis elegans Cul2 protein. Coimmunoprecipitation and chromatographic copurification data suggest that pVHL-Cul2 complexes exist in native cells. pVHL mutants that were unable to bind to complexes containing elongin C and Cul2 were likewise unable to inhibit the accumulation of hypoxia-inducible mRNAs. A model for the regulation of hypoxia-inducible mRNAs by pVHL is presented based on the apparent similarity of elongin C and Cul2 to Skp1 and Cdc53, respectively. These latter proteins form complexes that target specific proteins for ubiquitin-dependent proteolysis.


Isolation of an active form of the ATP + Mg2+-dependent protein phosphatase stimulated by the deinhibitor protein and by p-nitrophenyl phosphate.
A 75 kDa + 37 kDa-subunit high-Mr form of the ATP + Mg2+-dependent phosphatase, containing no modulator or inhibitor-1 and specific towards the beta-subunit of phosphorylase kinase, was isolated from dog liver. The phosphorylase phosphatase activity is stimulated by the deinhibitor protein, by p-nitrophenyl phosphate or by trypsin treatment. The sensitivity to modulator or inhibitor-1 can be increased dramatically by partial proteolysis. The enzyme inactivation by modulator after trypsin treatment can be reversed by protein kinase FA. The effects of p-nitrophenyl phosphate and the deinhibitor protein are synergistic, and p-nitrophenyl phosphate does not influence the sensitivity of the phosphatase to the heat-stable inhibitor proteins. These observations confirm that p-nitrophenyl phosphate stimulation is a suitable criterion for the identification of the active enzyme forms of the ATP + Mg2+-dependent phosphatase.


The crystal structure of Citrobacter freundii tyrosine phenol-lyase complexed with 3-(4'-hydroxyphenyl)propionic acid, together with site-directed mutagenesis and kinetic analysis, demonstrates that arginine 381 is required for substrate specificity.
The X-ray structure of tyrosine phenol-lyase (TPL) complexed with a substrate analog, 3-(4'-hydroxyphenyl)propionic acid, shows that Arg 381 is located in the substrate binding site, with the side-chain NH1 4.1 A from the 4'-OH of the analog. The structure has been deduced at 2.5 A resolution using crystals that belong to the P2(1)2(1)2 space group with a = 135.07 A, b = 143.91 A, and c = 59.80 A. To evaluate the role of Arg 381 in TPL catalysis, we prepared mutant proteins replacing arginine with alanine (R381A), with isoleucine (R381I), and with valine (R381V). The beta-elimination activity of R381A TPL has been reduced by 10(-4)-fold compared to wild type, whereas R381I and R381V TPL exhibit no detectable beta-elimination activity with L-tyrosine as substrate. However, R381A, R381I, and R381V TPL react with S-(o-nitrophenyl)-L-cysteine, beta-chloro-L-alanine, O-benzoyl-L-serine, and S-methyl-L-cysteine and exhibit k(cat) and k(cat)/Km values comparable to those of wild-type TPL. Furthermore, the Ki values for competitive inhibition by L-tryptophan and L-phenylalanine are similar for wild-type, R381A, and R381I TPL. Rapid-scanning-stopped flow spectroscopic analyses also show that wild-type and mutant proteins can bind L-tyrosine and form quinonoid complexes with similar rate constants. The binding of 3-(4'-hydroxyphenyl)propionic acid to wild-type TPL decreases at high pH values with a pKa of 8.4 and is thus dependent on an acidic group, possibly Arg404, which forms an ion pair with the analog carboxylate, or the pyridoxal 5'-phosphate Schiff base. R381A TPL shows only a small decrease in k(cat)/Km for tyrosine at lower pH, in contrast to wild-type TPL, which shows two basic pKas with an average value of about 7.8. Thus, it is possible that Arg 381 is one of the catalytic bases previously observed in the pH dependence of k(cat)/Km of TPL with L-tyrosine [Kiick, D. M., & Phillips. R. S. (1988) Biochemistry 27, 7333-7338], and hence Arg 381 is at least partially responsible for the substrate specificity of TPL.


Human papilloma virus in head and neck cancers-role and relevance in clinical management.
The biology and clinical behavior of Head and Neck Squamous Cell Carcinomas (HNSCCs) is very distinct within different subgroups due to the distinct molecular profiles for the HPV positive versus HPV negative tumors. HPV status is the most important independent prognostic variable in multivariate analysis taking into account all other prognostic factors like tumour stage, smoking status, age and performance status. The debate today is whether the intense therapy is too aggressive in this group of patients since they show a superior survival regardless of treatment strategies. A highly divergent prognosis and distinct biology of HPV positive and HPV negative HNSCCs underlines the fact that treating them as distinct diseases is the need of the hour. Infection with HPV is associated with less aggressive disease, better loco regional control and lower rates of second primary cancers. An important caveat that remains is the emergence of intermediate prognosis of HPV positive smokers and HPV negative non smokers. Though molecular biology has provided important data on the interaction of the HPV onco proteins with genes important in cell cycle control, also speculated to be involved in pathogenesis of HNSCC, more basic research is needed to describe the differential mechanisms of tumorigenesis among the HNSCCs that show presence and absence of HPV. This is clinically relevant to reduce morbidity without compromising tumour control in HPV positive patients and improving tumour control and co-morbid illness that could be pre-existing or treatment related in HPV negative patients. There may be a need for treatment intensification and incorporation of newer agents into induction chemotherapy protocols for the HPV negative patients and so HPV detection is important to aid in this selection. HPV tumour status is therefore more important than just providing the prognostic information in these classes of tumours. This article discusses the role and clinical relevance of HPV in HNSCCs.


Clinical documentation improvement for outpatients by implementing electronic medical records.
This observational study was conducted before and after implementing an electronic medical record (EMR) system to evaluate the change in outpatient workflow by implementation of EMR and the effectiveness of clinical documentation improvement (CDI). The number of hours for patient care increased by 89.2% (p < .05) and the hours for writing medical records after consulting decreased after implementation of EMR by 27.3% (p < .01). Implementation of EMR reduced nurses' workload to handle medical records by 78.8 (p < .05) but not changed for physicians. The necessary change in the information management process occurred after using the CDI indicator. We recommend that the "working hours of health professionals" and "handling hours for information resources" should be used widely as CDI indicators to improve workflow when implementing EMR.


Valuing Changes in Time Use in Low- and Middle-Income Countries.
Valuing changes in time use is often a critical element of economic analyses of development projects. In this paper we review the literature on the monetary value of time in low- and middle-income countries and find support for a commonly used benchmark of 50% of after-tax wages for time changes in activities in the informal sector, such as collecting water or traveling to health clinics. We offer recommendations to analysts who are conducting benefit-cost analyses in these settings about what methods they can use to estimate the value of time. These include a benefits transfer approach and also a relatively simple stated preference approach that might be deployed in a specific context if the project recommendation is sensitive to the assumption of the value of time or if the distribution of the benefits of time savings is especially important.


Biventricular repair of right atrial isomerism with complex congenital anomalies.
Biventricular repair is usually difficult to achieve in patients who have right atrial isomerism, which is typically associated with other complex cardiac anomalies. The procedure can be used in patients who have balanced ventricular structures. Herein, we report a successful surgical reconstruction, including biventricular repair, in a 4-year-old boy. The child's right atrial isomerism was associated with double-outlet right ventricle, a large atrial septal defect, a subaortic ventricular septal defect, valvular and infundibular pulmonary stenosis, left persistent superior vena cava, and hemiazygos continuation of an interrupted inferior vena cava. Balanced ventricles enabled biventricular repair, which we consider to be preferable to the Fontan procedure in such circumstances.


Novel AroA from Pseudomonas putida confers tobacco plant with high tolerance to glyphosate.
Glyphosate is a non-selective broad-spectrum herbicide that inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS, also designated as AroA), a key enzyme in the aromatic amino acid biosynthesis pathway in microorganisms and plants. Previously, we reported that a novel AroA (PpAroA1) from Pseudomonas putida had high tolerance to glyphosate, with little homology to class I or class II glyphosate-tolerant AroA. In this study, the coding sequence of PpAroA1 was optimized for tobacco. For maturation of the enzyme in chloroplast, a chloroplast transit peptide coding sequence was fused in frame with the optimized aroA gene (PparoA1(optimized)) at the 5' end. The PparoA1(optimized) gene was introduced into the tobacco (Nicotiana tabacum L. cv. W38) genome via Agrobacterium-mediated transformation. The transformed explants were first screened in shoot induction medium containing kanamycin. Then glyphosate tolerance was assayed in putative transgenic plants and its T(1) progeny. Our results show that the PpAroA1 from Pseudomonas putida can efficiently confer tobacco plants with high glyphosate tolerance. Transgenic tobacco overexpressing the PparoA1(optimized) gene exhibit high tolerance to glyphosate, which suggest that the novel PpAroA1 is a new and good candidate applied in transgenic crops with glyphosate tolerance in future.


Scleroderma and evidence based non-pharmaceutical treatment modalities for digital ulcers: a systematic review.
Digital ulcers are difficult to heal, increasing the chance of infection, gangrene, amputation and limited functional use of hands. They are a complication in scleroderma or systematic sclerosis (SSc) and occur in approximately 50% of patients. This is a systematic review of the evidence supporting the use of non-pharmaceutical therapeutic modalities and their effectiveness to facilitate the healing of chronic digital ulcers in patients with scleroderma. A comprehensive review of computerised databases from 2000-2013: PubMed/MEDLINE, CINAHL, Pedro, OT Seeker, OT Search, OVID, and Proquest as well as manual review of other resources was completed using the following search terms scleroderma or systemic sclerosis and/or digital ulcers, specific modalities (low level laser therapy, electrical stimulation, intermittent compression, ultrasound, vitamin E, myofascial release, wound dressings, iontophoresis, negative pressure therapy, and exercise), chronic wounds, and wound care. English language studies, from 2000 to January 2013, which used therapeutic modalities to facilitate healing of digital ulcers and use healing of the digital ulcer as an outcome measure were reviewed. Of the 403 identified articles, only 11 studies addressed non-pharmaceutical treatment modalities to facilitate healing for digital ulcers. Exercise had no direct effect on healing ulcers. The following studies were positive but have limitations in design and sample size:: hyperbaric oxygen therapy (n=2), negative pressure therapy (n=1), intermittent compression (n=27) and acoustic pressure wound healing (n=1). Vitamin E gel showed a significant difference compared to a control group (n=27). Iontophoresis studies have shown that the modality increases blood flow but the results in five different studies are mixed and the application and intensity were inconsistent. No one modality was proven to be the most effective. Larger efficacy studies on treating digital ulcers are needed in order to develop appropriate care guidelines to improve outcomes, promote function and lower health-care costs.


Therapeutic Advances and Treatment Options in Metastatic Melanoma.
Over the past several years, management of advanced melanoma has been transformed by the development and approval of novel therapeutic approaches. Genetically targeted therapies are now effective treatment options for the approximately 50% of patients whose melanomas harbor activating point mutations in BRAF. Combination regimens of small-molecule inhibitors have been developed to delay the onset of acquired resistance. Specifically, combined BRAF and MEK inhibition improves response rates and survival compared with single-agent BRAF inhibitors and has now received regulatory approval. During the same time frame, excitement has surrounded the development of immunotherapy with checkpoint inhibitors. New immune checkpoint inhibitors blocking cytotoxic T lymphocyte antigen-4 (CTLA4) or programmed death-1 receptor/ligand (PD-1/PD-L1) improve patient outcomes by promoting an antitumor immune response. These agents have been associated with an increasing number of durable responses and are being developed in various combinations. In this review, we discuss the development of these targeted and immune therapies, review current patient management, and highlight future directions.


Left ventricular unloading during veno-arterial ECMO: a review of percutaneous and surgical unloading interventions.
Short-term mechanical support by veno-arterial extracorporeal membrane oxygenation (VA ECMO) is more and more applied in patients with severe cardiogenic shock. A major shortcoming of VA ECMO is its variable, but inherent increase of left ventricular (LV) mechanical load, which may aggravate pulmonary edema and hamper cardiac recovery. In order to mitigate these negative sequelae of VA ECMO, different adjunct LV unloading interventions have gained a broad interest in recent years. Here, we review the whole spectrum of percutaneous and surgical techniques combined with VA ECMO reported to date.


Cancer stem cells in colorectal cancer: a review.
Colorectal cancer (CRC) is the second most common cancer in women and the third most common in men. Adenocarcinoma accounts for 90% of CRC cases. There has been accumulating evidence in support of the cancer stem cell (CSC) concept of cancer which proposes that CSCs are central in the initiation of cancer. CSCs have been the focus of study in a range of cancers, including CRC. This has led to the identification and understanding of genes involved in the induction and maintenance of pluripotency of stem cells, and markers for CSCs, including those investigated specifically in CRC. Knowledge of the expression pattern of CSCs in CRC has been increasing in recent years, revealing a heterogeneous population of cells within CRC ranging from pluripotent to differentiated cells, with overlapping and sometimes unique combinations of markers. This review summarises current literature on the understanding of CSCs in CRC, including evidence of the presence of CSC subpopulations, and the stem cell markers currently used to identify and localise these CSC subpopulations. Future research into this field may lead to improved methods for early detection of CRC, novel therapy and monitoring of treatment for CRC and other cancer types.


Thyroid dysfunction in patients with type 2 diabetes mellitus in Jordan.
To investigate the prevalence of thyroid dysfunction and autoimmunity in type 2 diabetic patients. The study was conducted at the National Center for Diabetes, Endocrinology and Genetics, Jordan University Hospital, Amman, Jordan, between March 2000 and September 2000. A group of 908 type 2 diabetic patients (T2DM) were recruited in the study and underwent investigations for thyroid functions; free thyroxine (FT4), free tri-iodothyronine (FT3) and thyroid stimulating hormone (TSH). Six hundred had performed thyroid autoantibodies, thyroid peroxidase antibodies (TPOab) or antimicrosomal antibodies (AMA) and thyroglobulin antibodies (Tgab). They were compared with 304 non-diabetics, of those 282 had performed thyroid antibodies. Fifty-three (5.9%) of diabetic patients were known to have thyroid disease. As a direct result of screening, new thyroid disease cases were diagnosed in 6.6% of the patients. Thus, the overall prevalence of thyroid disease was found to be 12.5%. The most common was subclinical hypothyroidism (4.1%). In the control group, the prevalence of thyroid disease was 6.6%. The most common was subclinical hypothyroidism (5%). There was a significant difference between diabetics and control subjects p=0.0064. Positive TPOab was found in 8.3% of T2DM patients (N=600) versus 10.3% in the control group (N=282) p=0.412. Positivity for both TPOab and Tgab was found to be 2.5% of T2DM versus 6% of the control subjects p=0.0155. This study suggests that diabetic patients should be screened for asymptomatic thyroid dysfunction.


Acceptability, Feasibility, and Quality of Telehealth for Adolescent Health Care Delivery During the COVID-19 Pandemic: Cross-sectional Study of Patient and Family Experiences.
Data regarding the acceptability, feasibility, and quality of telehealth among adolescents and young adults (AYA) and their parents and caregivers (caregivers) are lacking. The aim of this study was to assess the noninferiority of telehealth versus in-person visits by comparing acceptability with respect to efficiency, effectiveness, equity, patient-centeredness, and confidentiality. Cross-sectional web-based surveys were sent to caregivers and AYA following video visits within an Adolescent Medicine subspecialty clinic in May-July 2020. Proportions of AYA and caregivers who rated telehealth as noninferior were compared using chi-squared tests. Feasibility was assessed via items measuring technical difficulties. Deductive thematic analysis using the Institute of Medicine dimensions of health care quality was used to code open-ended question responses. Survey response rates were 20.5% (55/268) for AYA and 21.8% (123/563) for caregivers. The majority of the respondents were White cisgender females. Most AYA and caregivers rated telehealth as noninferior to in-person visits with respect to confidentiality, communication, medication management, and mental health care. A higher proportion of AYA compared to caregivers found telehealth inferior with respect to confidentiality (11/51, 22% vs 3/118, 2.5%, P<.001). One-quarter (14/55) of the AYA patients and 31.7% (39/123) of the caregivers reported technical difficulties. The dominant themes in the qualitative data included advantages of telehealth for efficiency and equity of health care delivery. However, respondents' concerns included reduced safety and effectiveness of care, particularly for patients with eating disorders, owing to lack of hands-on examinations, collection of vital signs, and laboratory testing. Telehealth was highly acceptable among AYA and caregivers. Future optimization should include improving privacy, ameliorating technical difficulties, and standardizing at-home methods of obtaining patient data to assure patient safety.


Ambulatory monitoring of pulmonary artery pressure. A preliminary clinical evaluation.
Traditional measurement and recording methods are inadequate for continuous monitoring of ambulatory pulmonary artery pressure. Therefore a new miniaturised solid state system has been developed and assessed. A manometer tipped catheter, inserted via a subclavian or cephalic vein, was used together with an isolated amplifier and peak detectors to determine systolic and diastolic pressures. Pressures were averaged over 30 seconds and stored in digital memory. After a 24 hour recording period data were rapidly transferred to a microcomputer for numerical or graphical display. Thirteen patients had continuous ambulatory monitoring performed for between 24 and 96 hours, in seven to evaluate symptoms of dyspnoea in subjects with valvular or coronary disease (group 1), and in six to achieve optimal oral treatment for left heart failure (group 2). The catheter was calibrated before insertion and was rechecked after removal. There was less than 1% zero level drift and similar gain stability. Systolic pressures ranged from 10 to 97 (mean 39.5) mmHg, and diastolic from 1 to 46 (mean 15.3) mmHg. Four patients in group 1 had symptoms of dyspnoea associated with normal pressures, while three had raised pressures. Four of the six patients monitored in group 2 had major alterations in their treatment based on data obtained during monitoring. There were no complications. This system, which allows safe, reliable, and prolonged recording of ambulatory pulmonary artery pressure, represents a considerable advance in the ability to assess the cause of dyspnoea and to manage left heart failure.


An efficient algorithm for automatically generating multivariable fuzzy systems by Fourier series method.
By exploiting the Fourier series expansion, we have developed a new constructive method of automatically generating a multivariable fuzzy inference system from any given sample set with the resulting multivariable function being constructed within any specified precision to the original sample set. The given sample sets are first decomposed into a cluster of simpler sample sets such that a single input fuzzy system is constructed readily for a sample set extracted directly from the cluster independent of the other variables. Once the relevant fuzzy rules and membership functions are constructed for each of the variables completely independent of the other variables, the resulting decomposed fuzzy rules and membership functions are integrated back into the fuzzy system appropriate for the original sample set requiring only a moderate cost of computation in the required decomposition and composition processes. After proving two basic theorems which we need to ensure the validity of the decomposition and composition processes of the system construction, we have demonstrated a constructive algorithm of a multivariable fuzzy system. Exploiting an implicit error bound analysis available at each of the construction steps, the present Fourier method is capable of implementing a more stable fuzzy system than the power series expansion method of ParNeuFuz and PolyNeuFuz, covering and implementing a wider range of more robust applications.


Cholagogic effect of ginger and its active constituents.
The effect of bile secretion in rats was examined in order to clarify the stomachic action of ginger and also to investigate its active constituents. The results showed that mainly the acetone extracts of ginger, which contain essential oils and pungent principles, caused an increase in the bile secretion. Further analyses for the active constituents of the acetone extracts through column chromatography indicated that [6]-gingerol and [10]-gingerol, which are the pungent principles, are mainly responsible for the cholagogic effect of ginger.


Selective impairment of slow axonal transport after optic nerve injury in adult rats.
To investigate cellular responses of injured mammalian CNS neurons, we examined the slow transport of cytoskeletal proteins in rat retinal ganglion cell (RGC) axons within the ocular stump of optic nerves that were crushed intracranially. RGC proteins were labeled by an intravitreal injection of 35S-methionine, and optic nerves were examined by SDS PAGE at different times after injury. In one group of rats, the RGC proteins were labeled 1 week after crushing. From 14 to 67 d after axotomy, the labeling of tubulin and neurofilaments was reduced in relation to other labeled proteins and to the labeling of tubulin and neurofilaments in the intact optic nerve of controls. To determine whether this reduction in labeling was due to an alteration in axonal transport after axotomy, we prelabeled RGC proteins 1 week before crushing. In such experiments, the rate of slow axonal transport of tubulin and neurofilaments decreased approximately 10-fold from 6 to 60 d after injury. Our results cannot be due only to the retrograde degeneration of RGCs and injured axons caused by axotomy in the optic nerve, because fast axonal protein transport and the fluorescent labeling of many axons were preserved in the ocular stumps of these optic nerves. This selective failure of the slow axonal transport of tubulin and neurofilaments may affect the renewal of the cytoskeleton and contribute to the gradual degeneration of RGCs that is observed after axotomy. The alterations in slow transport we document here differ from the enhanced rates we previously reported when injured RGC axons regenerated along peripheral nerve segments grafted to the ocular stump of transected optic nerves (McKerracher et al., 1990).


Headspace solid-phase microextraction combined with mass spectrometry as a powerful analytical tool for profiling the terpenoid metabolomic pattern of hop-essential oil derived from Saaz variety.
Hop (Humulus lupulus L., Cannabaceae family) is prized for its essential oil contents, used in beer production and, more recently, in biological and pharmacological applications. In this work, a method involving headspace solid-phase microextraction and gas chromatography-mass spectrometry was developed and optimized to establish the terpenoid (monoterpenes and sesquiterpenes) metabolomic pattern of hop-essential oil derived from Saaz variety as a mean to explore this matrix as a powerful biological source for newer, more selective, biodegradable and naturally produced antimicrobial and antioxidant compounds. Different parameters affecting terpenoid metabolites extraction by headspace solid-phase microextraction were considered and optimized: type of fiber coatings, extraction temperature, extraction time, ionic strength, and sample agitation. In the optimized method, analytes were extracted for 30 min at 40°C in the sample headspace with a 50/30 μm divinylbenzene/carboxen/polydimethylsiloxane coating fiber. The methodology allowed the identification of a total of 27 terpenoid metabolites, representing 92.5% of the total Saaz hop-essential oil volatile terpenoid composition. The headspace composition was dominated by monoterpenes (56.1%, 13 compounds), sesquiterpenes (34.9%, 10), oxygenated monoterpenes (1.41%, 3), and hemiterpenes (0.04%, 1) some of which can probably contribute to the hop of Saaz variety aroma. Mass spectrometry analysis revealed that the main metabolites are the monoterpene β-myrcene (53.0 ± 1.1% of the total volatile fraction), and the cyclic sesquiterpenes, α-humulene (16.6 ± 0.8%), and β-caryophyllene (14.7 ± 0.4%), which together represent about 80% of the total volatile fraction from the hop-essential oil. These findings suggest that this matrix can be explored as a powerful biosource of terpenoid metabolites.


Low-level exposure to lead and atherosclerosis in the carotid arteries: Results from the Swedish population-based cohort SCAPIS.
Lead exposure is associated with cardiovascular disease. Atherosclerosis has been hypothesized to be one of the underlying mechanisms behind this association. To investigate whether lead exposure is associated with an increased risk of atherosclerosis in the carotid arteries in a large Swedish population-based cohort. We performed a cross-sectional study using data from the population-based Swedish CardioPulmonary bioImage Study (SCAPIS), including 5622 middle-aged men and women, enrolled 2013-2018. Blood lead (B-Pb), measured by inductively coupled plasma mass spectrometry, was used as exposure biomarker. The presence of atherosclerotic plaque in the carotid arteries (yes/no), total plaque area (mm2) and the presence of large plaques (>25 mm2) were determined by ultrasonography. Associations between B-Pb and the different outcomes were analysed using Poisson and linear regression models, adjusted for potential confounders. Atherosclerotic plaque was present in 57% of the individuals, for whom the median total plaque area was 16 mm2 (range: 0.2-222). The median B-Pb concentration was 14 μg/L (range: 0.75-203). After adjusting for potential confounders, individuals in the fourth quartile of B-Pb (Q4) had a prevalence ratio (PR) for plaque of 1.08 (95% CI: 1.01, 1.16) when compared with the first quartile (Q1). A 10 μg/L increase in B-Pb concentrations was associated with an increase of 0.92 mm2 (95% CI: 0.14, 1.71) in total plaque area. The PR for large plaque was 1.09 (95% CI: 0.84, 1.42 for Q4 vs Q1). This study shows an association between B-Pb and atherosclerosis in the carotid arteries providing some support for the hypothesis that atherosclerosis is one of the mechanisms underlying the association between lead exposure and cardiovascular disease.


Comparison of two methods forecasting binding rate of plasma protein.
By introducing the descriptors calculated from the molecular structure, the binding rates of plasma protein (BRPP) with seventy diverse drugs are modeled by a quantitative structure-activity relationship (QSAR) technique. Two algorithms, heuristic algorithm (HA) and support vector machine (SVM), are used to establish linear and nonlinear models to forecast BRPP. Empirical analysis shows that there are good performances for HA and SVM with cross-validation correlation coefficients Rcv(2) of 0.80 and 0.83. Comparing HA with SVM, it was found that SVM has more stability and more robustness to forecast BRPP.


Electron densities inferred from plasma wave spectra obtained by the Waves instrument on Van Allen Probes.
The twin Van Allen Probe spacecraft, launched in August 2012, carry identical scientific payloads. The Electric and Magnetic Field Instrument Suite and Integrated Science suite includes a plasma wave instrument (Waves) that measures three magnetic and three electric components of plasma waves in the frequency range of 10 Hz to 12 kHz using triaxial search coils and the Electric Fields and Waves triaxial electric field sensors. The Waves instrument also measures a single electric field component of waves in the frequency range of 10 to 500 kHz. A primary objective of the higher-frequency measurements is the determination of the electron density ne at the spacecraft, primarily inferred from the upper hybrid resonance frequency fuh. Considerable work has gone into developing a process and tools for identifying and digitizing the upper hybrid resonance frequency in order to infer the electron density as an essential parameter for interpreting not only the plasma wave data from the mission but also as input to various magnetospheric models. Good progress has been made in developing algorithms to identify fuh and create a data set of electron densities. However, it is often difficult to interpret the plasma wave spectra during active times to identify fuh and accurately determine ne . In some cases, there is no clear signature of the upper hybrid band, and the low-frequency cutoff of the continuum radiation is used. We describe the expected accuracy of ne and issues in the interpretation of the electrostatic wave spectrum.


Multiple factors contribute to bimodal toxin gene expression in Clostridioides (Clostridium) difficile.
Clostridioides (formerly Clostridium) difficile produces two major toxins, TcdA and TcdB, upon entry into stationary phase. Transcription of tcdA and tcdB requires the specialized sigma factor, σTcdR , which also directs RNA Polymerase to transcribe tcdR itself. We fused a gene for a red fluorescent protein to the tcdA promoter to study toxin gene expression at the level of individual C. difficile cells. Surprisingly, only a subset of cells became red fluorescent upon entry into stationary phase. Breaking the positive feedback loop that controls σTcdR production by engineering cells to express tcdR from a tetracycline-inducible promoter resulted in uniform fluorescence across the population. Experiments with two regulators of tcdR expression, σD and CodY, revealed neither is required for bimodal toxin gene expression. However, σD biased cells toward the Toxin-ON state, while CodY biased cells toward the Toxin-OFF state. Finally, toxin gene expression was observed in sporulating cells. We conclude that (i) toxin production is regulated by a bistable switch governed by σTcdR , which only accumulates to high enough levels to trigger toxin gene expression in a subset of cells, and (ii) toxin production and sporulation are not mutually exclusive developmental programs.


Profile of patients with gastrointestinal stromal tumors (GIST).
There is an improvement on the GIST treatment in last decade due to biomolecular research and adjuvant therapy with tyrosine kinases inibitors. However, both modalities of treatment rarely are available in Brazilian public hospital. Evaluate GIST patients profile in public oncologic hospital. A retrospective study was made on patients with GIST diagnosed and treated between 2001 and 2013. Sixty-nine patients were included, mean age 59 years with slight predominance in females (51%). The main symptom was abdominal pain associated with incidental imaging finding. The occurrence of other associated neoplasm was in 28.8% of cases. The positivity of CD117 was 97.1%. The most frequent location was the stomach in 55.1% of cases. The R0 resection was possible in 63.8% and the recurrence rate was 20.3 %, with liver and peritoneum the main affected sites. Overall survival in the whole sample was 71%. Free survival rate of disease was 64%. The use of imatinib was limited to patients with residual disease (unresectable disease, R2 and R1 resection), metastatic disease or recurrence. In order to improve GIST treatment is necessary to add the biomolecular analysis to risk stratification. However, for this to occur, incentive in biomolecular research is required, to increase the possibility of patient survival.


Frontal Hyperostosis in the Patients of Craniosynostosis After Cranial Distraction Osteogenesis.
Conventional fronto-orbital advancement and distraction osteogenesis (DOG) have been used to treat craniosynostosis, both of which are considered effective. During the authors' practice, a phenomenon of frontal hyperostosis has been observed in the patients of craniosynostosis after DOG, which has yet to be reported in the literature. The purpose of this study is trying to identify the factors related to the phenomenon. From 1997 to 2010, all patients of craniosynostosis undergoing DOG were reviewed. The patient's age at operation, consolidation period, numbers of distractor, distance of distraction, and duration from removal of the distractors to identification of the phenomenon on computed tomography were recorded. The phenomenon was considered positive when the hyperostosis appeared on the frontal bone, where it was neither the osteotomy site nor the previous position of distractor. A total of 61 patients were included in this study, including 26 syndromic and 35 nonsyndromic patients. Two syndromic and 6 nonsyndromic patients had the phenomenon. There was no statistical difference between the patients with and without the phenomenon in comparison with the age, number of the distractor, consolidation period, and the distance of distraction. Frontal hyperostosis happened in some patients of craniosynostosis after DOG. Although no significant difference was demonstrated, the incidence of hyperostosis was higher in nonsyndromic patients and the patients of hyperostosis had shorter distance of distraction in both syndromic and nonsyndromic groups. Although the definite cause was unknown, we should pay attention to the phenomenon after distraction.


Androgen dynamics in vitro in the human prostate gland. Effect of oestradiol-17 .
Normal, hyperplastic and adenocarcinomatous human prostatic tissue was perfused in vitro with radioactively labelled androstenedione, testosterone and 5alpha-dihydrotestosterone with and without added oestradiol-17beta. Various parameters of tissue-steroid relationship were measured at the steady state. When oestradiol (0.11 or 0.22mumol/l) was added to the perfusing medium, the entry of the steroids into the tissue and their metabolism was increased in the majority of the glands studied. The ;uptake' of all the steroids varied, in response to the addition of oestradiol, in both normal and adenocarcinomatous glands in a way differing from the response of hyperplastic glands. As a consequence, the tissue clearance of the steroids, particularly of androstenedione and testosterone, increased in normal and adenocarcinomatous glands in the presence of oestradiol, and decreased in the hyperplastic tissues. At a concentration 0.33mumol/l, oestradiol decreased the entry of the steroids in all the tissues studied, while the clearance of steroids tended to decrease. The significance of these findings in terms of the regulation of androgen dynamics in vivo in the normal and diseased human prostate, with particular regard to the response to oestrogen treatment, is discussed.


Optimal Multivariate Tuning with Neuron-Level and Population-Level Energy Constraints.
Optimality principles have been useful in explaining many aspects of biological systems. In the context of neural encoding in sensory areas, optimality is naturally formulated in a Bayesian setting as neural tuning which minimizes mean decoding error. Many works optimize Fisher information, which approximates the minimum mean square error (MMSE) of the optimal decoder for long encoding time but may be misleading for short encoding times. We study MMSE-optimal neural encoding of a multivariate stimulus by uniform populations of spiking neurons, under firing rate constraints for each neuron as well as for the entire population. We show that the population-level constraint is essential for the formulation of a well-posed problem having finite optimal tuning widths and optimal tuning aligns with the principal components of the prior distribution. Numerical evaluation of the two-dimensional case shows that encoding only the dimension with higher variance is optimal for short encoding times. We also compare direct MMSE optimization to optimization of several proxies to MMSE: Fisher information, maximum likelihood estimation error, and the Bayesian Cramér-Rao bound. We find that optimization of these measures yields qualitatively misleading results regarding MMSE-optimal tuning and its dependence on encoding time and energy constraints.


Cystic disease of the renal medulla associated with retinitis pigmentosa and imino acid abnormalities.
A 22 year old man with renal failure associated with salt wasting, retinitis pigmentosa, and imino acid abnormalities is reported. Renal tissue showed changes compatible with medullary cystic disease or juvenile nephronophtisis. Special studies were carried out to clarify the mechanism of renal salt wasting. Retinal pigmentary changes were found in three siblings and a male cousin; in two siblings studied abnormal retinal function was demonstrated. the parents who were first cousins once removed were normal. Fifty kindred members were examined and none showed unequivocal signs of renal disease. Autosomal recessive transmission of ocular and renal disease appeared likely. The separation of medullary cystic disease from juvenile nephronophthisis on the basis of onset and mode of inheritance is probably artificial.


Chemical compositions, antioxidant and antimicrobial activities of essential oils of Piper caninum Blume.
Chemical composition, antioxidant and antimicrobial activities of the fresh leaves and stems oils of Piper caninum were investigated. A total of forty eight constituents were identified in the leaves (77.9%) and stems (87.0%) oil which were characterized by high proportions of phenylpropanoid, safrole with 17.1% for leaves and 25.5% for stems oil. Antioxidant activities were evaluated by using β-carotene/linoleic acid bleaching, DPPH radical scavenging and total phenolic content. Stems oil showed the highest inhibitory activity towards lipid peroxidation (114.9 ± 0.9%), compared to BHT (95.5 ± 0.5%), while leaves oil showed significant total phenolic content (27.4 ± 0.5 mg GA/g) equivalent to gallic acid. However, the essential oils showed weak activity towards DPPH free-radical scavenging. Evaluation of antimicrobial activity revealed that both oils exhibited strong activity against all bacteria strains with MIC values in the range 62.5 to 250 μg/mL, but weak activity against fungal strains. These findings suggest that the essential oils can be used as antioxidant and antimicrobial agents for therapeutic, nutraceutical industries and food manufactures.


Reconstructive Surgery of Inguinal Defects: A Systematic Literature Review of Surgical Etiology and Reconstructive Technique.
This study aimed to evaluate the literature regarding surgical etiology demanding inguinal reconstructive surgery, associated reconstructive techniques and outcomes. A systematic literature search was performed according to the PRISMA statement between 1996-2016. A total of 64 articles were included, comprising 816 patients. Two main subgroups of patients were identified: Oncological resections (n=255, 31%), and vascular surgery (n=538, 66%). Oncological resection inguinal defects were treated with pedicled myocutaneous flaps (n=166, 65%), fasciocutaneous flaps (77, 31%), muscle flaps (7, 3%) and direct closure (3, 1%). Vascular surgery complications were treated with muscle flaps (n=513, 95%). Complications for the respective subgroup (oncological resections, vascular surgery) were: infection (24%, 14%), seroma (34%, 7.5%), flap dehiscence/delayed healing (20.6%, 40.8%,). The total reintervention rate was 20%. Reconstruction of inguinal defects should be addressed on a case-by-case basis. Myocutaneous flaps were favoured after oncological resections, while muscle flaps were preferred after vascular surgery.


Immunoglobulin synthesis in the "resting" breast.
Nulliparous women have a greater risk of developing breast cancer than women who have borne children, but so far no functional differences in breast tissue have been reported between parous women and nulliparae. Macroscopically and histologically normal breast tissue was obtained from 74 women of reproductive age during biopsy of benign breast lesions and was examined for the presence of plasma cells by immunfluorescence. Immunoglobulin synthesis was detected by an in-vitro culture technique. Synthesis of IgA was detected in 81% of specimans of IgG in 45%, and of IgM in 3%. IgA synthesis much more intense than IgG or IgM synthesis. Plasma cells containing IgA were seen in 71% of the specimens examined, and 88% of specimens had deposits of IgA in the ductules. The findings were not significantly incluenced by the nature of the condition necessitating biopsy or by oral contraception. Nulliparous women showed no cyclical changes, but among parous women IgA synthesis was more intense during luteal phase of the menstrual cycle. This suggests that after the first pregnancy the breast is more sensitive to progesterone.


Pharmacophoric requirements for the cannabinoid side chain. Probing the cannabinoid receptor subsite at C1'.
Earlier work from our laboratories has provided evidence for the existence of a subsite within the CB1 and CB2 cannabinoid receptor binding domain corresponding to substituents at the benzylic side chain position of classical cannabinoids. The existence and stereochemical features of this subsite have now been probed through the synthesis of a novel series of (-)-Delta(8)-tetrahydrocannabinol analogues bearing C1'-ring substituents. Of the compounds described here, those with C1'-dithiolane (1c), C1'-dioxolane (2d), and cyclopentyl (2a) substituents exhibited the highest affinities for CB1 and CB2. We used molecular modeling approaches to better define the stereochemical limits of the putative subsite. In vitro pharmacological testing found 1c to be a potent CB1 agonist.


Cross-platform prediction of gene expression signatures.
Gene expression signatures can predict the activation of oncogenic pathways and other phenotypes of interest via quantitative models that combine the expression levels of multiple genes. However, as the number of platforms to measure genome-wide gene expression proliferates, there is an increasing need to develop models that can be ported across diverse platforms. Because of the range of technologies that measure gene expression, the resulting signal values can vary greatly. To understand how this variation can affect the prediction of gene expression signatures, we have investigated the ability of gene expression signatures to predict pathway activation across Affymetrix and Illumina microarrays. We hybridized the same RNA samples to both platforms and compared the resultant gene expression readings, as well as the signature predictions. Using a new approach to map probes across platforms, we found that the genes in the signatures from the two platforms were highly similar, and that the predictions they generated were also strongly correlated. This demonstrates that our method can map probes from Affymetrix and Illumina microarrays, and that this mapping can be used to predict gene expression signatures across platforms.


Targeting the fatty acid biosynthesis enzyme, beta-ketoacyl-acyl carrier protein synthase III (PfKASIII), in the identification of novel antimalarial agents.
The importance of fatty acids to the human malaria parasite, Plasmodium falciparum, and differences due to a type I fatty acid synthesis (FAS) pathway in the parasite, make it an attractive drug target. In the present study, we developed and a utilized a pharmacophore to select compounds for testing against PfKASIII, the initiating enzyme of FAS. This effort identified several PfKASIII inhibitors that grouped into various chemical classes of sulfides, sulfonamides, and sulfonyls. Approximately 60% of the submicromolar inhibitors of PfKASIII inhibited in vitro growth of the malaria parasite. These compounds inhibited both drug sensitive and resistant parasites and testing against a mammalian cell line revealed an encouraging in vitro therapeutic index for the most active compounds. Docking studies into the active site of PfKASIII suggest a potential binding mode that exploits amino acid residues at the mouth of the substrate tunnel.


Electrochemical nonenzymatic glucose sensors catalyzed by Au nanoclusters on metallic nanotube arrays and polypyrrole nanowires.
Monitoring blood glucose level is critical, since its abnormality leads to diabetes and causes death, even though glucose is essential for human living. Herein, the sensing study was performed on electrochemical nonenzymatic glucose sensors, which are composed of an Au nanocluster (AuNC) catalyst deposited on a metallic nanotube array (MeNTA) and polypyrrole nanowire (PPyNW). The AuNC was produced by irradiating a femtosecond pulse laser to the Au precursor solution, and it is a simple and facile method. The successful deposition of AuNC on both MeNTA and PPyNW was confirmed by means of the surface morphology and the Au content increase. On the exploration by cyclic voltammetry in alkaline condition, AuNC/MeNTA electrodes showed better performance than AuNC/PPyNW electrodes: The former was a remarkable electrocatalytic detector towards glucose oxidation with better sensitivity, lower detection limit, wider linear range, and longer-term stability without interference from potential interfering agents such as ascorbic acid, urea, NaCl, KCl, etc. Moreover, nonenzymatic AuNC/MeNTA electrodes exhibited high precision and accuracy in real human blood samples and, thus, can be a promising candidate in glucose sensing applications.


An Ultrasound Classification of Anterior Talofibular Ligament (ATFL) Injury.
To develop a classification of ATFL injury based on the ultrasonography. The ultrasound images of 560 cases that had chronic ankle instability were studied from May 2012 to May 2015. All the patients accepted ultrasonography to type the ATFL injury. The ATFL injuries could be divided into six subtypes based on ultrasound imaging: I: Intact ligament; II: Injury at the fibular side; III:Injury at the talar side; IV: Tear at the midsubstance; V:Ligament absorbed; VI:Combined injury. Ultrasound could be used to evaluate and type the injury of ATFL. This kind of classification could be helpful in the preoperative decision of ATFL procedure.


Semaphorin-5A maintains epithelial phenotype of malignant pancreatic cancer cells.
Pancreatic cancer (PC) is a highly aggressive disease, and the lethality of this disease stems from early metastatic dissemination where surgical removal cannot provide a cure. Improvement of the therapeutic outcome and overall survival of PC patients requires to understand the fundamental processes that lead to metastasis such as the gain of cellular migration ability. One such family of proteins, which are essential players of cellular migration, is Semaphorin. Previously, we have identified one of the Semaphorin family member, Semaphorin-5A (SEMA5A) to be involved in organ-specific homing during PC metastasis. We have also demonstrated that SEMA5A has a constitutive expression in PC cell lines derived from metastatic sites in comparison with low endogenous expression in the primary tumor-derived cell line. In this study, we examined whether constitutive SEMA5A expression in metastatic PC cells regulates tumor growth and metastatic potential. We generated SEMA5A knockdown in T3M-4 and CD18/HPAF cells and assessed their phenotypes on in vitro motility, tumor growth, and metastatic progression. In contrary to our initial expectations, orthotopic injection of SEMA5A knockdown cells into nude mice resulted in a significant increase in both tumor burden and liver metastases in comparison with the Control cells. Similarly, we observed higher in vitro migratory potential with pronounced morphological changes associated with epithelial-mesenchymal transition (EMT), a decrease in the expression of epithelial marker E-cadherin (E-Cad), increase in the expression of mesenchymal markers N-cadherin (N-Cad) and Snail and the activation of the Wnt-signaling pathway in SEMA5A knockdown cells. Furthermore, re-establishing SEMA5A expression with a knockdown resistant mouse Sema5A in SEMA5A knockdown cells resulted in a reversion to the epithelial state (mesenchymal-epithelial transition; MET), as indicated by the rescue of E-Cad expression and a decrease in N-Cad and Snail expression. Collectively, our data suggest that SEMA5A expression maintains epithelial phenotype in the metastatic microenvironment.


Breaking Continuous Flash Suppression: A New Measure of Unconscious Processing during Interocular Suppression?
Until recently, it has been thought that under interocular suppression high-level visual processing is strongly inhibited if not abolished. With the development of continuous flash suppression (CFS), a variant of binocular rivalry, this notion has now been challenged by a number of reports showing that even high-level aspects of visual stimuli, such as familiarity, affect the time stimuli need to overcome CFS and emerge into awareness. In this "breaking continuous flash suppression" (b-CFS) paradigm, differential unconscious processing during suppression is inferred when (a) speeded detection responses to initially invisible stimuli differ, and (b) no comparable differences are found in non-rivalrous control conditions supposed to measure non-specific threshold differences between stimuli. The aim of the present study was to critically evaluate these assumptions. In six experiments we compared the detection of upright and inverted faces. We found that not only under CFS, but also in control conditions upright faces were detected faster and more accurately than inverted faces, although the effect was larger during CFS. However, reaction time (RT) distributions indicated critical differences between the CFS and the control condition. When RT distributions were matched, similar effect sizes were obtained in both conditions. Moreover, subjective ratings revealed that CFS and control conditions are not perceptually comparable. These findings cast doubt on the usefulness of non-rivalrous control conditions to rule out non-specific threshold differences as a cause of shorter detection latencies during CFS. Thus, at least in its present form, the b-CFS paradigm cannot provide unequivocal evidence for unconscious processing under interocular suppression. Nevertheless, our findings also demonstrate that the b-CFS paradigm can be fruitfully applied as a highly sensitive device to probe differences between stimuli in their potency to gain access to awareness.


Effects of HIV on executive function and verbal fluency in Cameroon.
HIV-associated neurocognitive disorders (HAND) are frequently associated with impaired executive function and verbal fluency. Given limited knowledge concerning HAND in Sub-Saharan-Africa and lack of Cameroonian adult neuropsychological (NP) test norms, we administered four executive function [Halstead Category Test (HCT), Wisconsin Card Sorting Test (WCST), Color Trails-II (CTT2), and Stroop Color-Word-Interference (SCWT)] and three verbal fluency (Category, Action, and Letter Fluency) tests to 742 adult Cameroonians (395 HIV-, 347 HIV+). We developed demographically-corrected NP test norms and examined the effects of HIV and related variables on subjects' executive function and verbal fluency. HIV+ subjects had significantly lower T-scores on CTT2 (P = 0.005), HCT (P = 0.032), WCST (P < 0.001); lower executive function composite (P = 0.002) and Action Fluency (P = 0.03) T-scores. ART, viremia, and CD4 counts did not affect T-scores. Compared to cases harboring other viral subtypes, subjects harboring HIV-1 CRF02_AG had marginally higher CTT2 T-scores, significantly higher SCWT (P = 0.015) and executive function (P = 0.018) T-scores. Thus, HIV-1 infection in Cameroon is associated with impaired executive function and some aspects of verbal fluency, and viral genotype influenced executive function. We report the first normative data for assessing executive function and verbal fluency in adult Cameroonians and provide regression-based formulas for computing demographically-adjusted T-scores. These norms will be useful for investigating HIV/AIDS and other diseases affecting cognitive functioning in Cameroon.


[Salmonella serotypes identified in Mexican health services].
To identify the different Salmonella strain serotypes isolated at public and private laboratories in Mexico and at the Institute for Epidemiologic Diagnosis and Referral (InDRE). A total of 24,394 Salmonella strains collected from 1972 to 1999 in public and private health laboratories of Mexico were analyzed with the Kauffmann-White method, using antisera produced by InDRE, according to Centers for Disease Control and Prevention (CDC, Atlanta, GA) standards; 15,843 (64.9%) samples were from human sources and 8,551 (35.1%) from non-human sources. One hundred ninety nine different serotypes were identified. The most frequent serotype in human beings was S. Typhimurium (20.4%), followed by S. Enteritidis (18.3%). In the past few years, the frequency of S. Enteritidis has been increasing, surpassing that of S. Typhimurium since 1991. Presently S. Enteritidis is the most frequently isolated serotype. In non-human sources, S. Derby (13.8%) and S. Anatum (8.5%) are the most frequent strains. Salmonella serotypes most frequently isolated in Mexico are: S. Typhimurium, S. Enteritidis, S. Derby, S. Agona y S. Anatum. From the epidemiologic standpoint, it is necessary to identify circulating and emerging Salmonella serotypes in order to target pertinent preventative interventions.


Regulation of apoptosis signal-regulating kinase 1 in redox signaling.
Apoptosis signal-regulating kinase 1 (ASK1) is a member of the mitogen-activated protein kinase (MAPK) kinase kinase family and elicits a wide variety of cellular responses to various types of stress through activation of the JNK and p38 MAPK pathways. ASK1 is preferentially activated in response to oxidative stress, but this regulatory mechanism is still not completely understood. In our previous report, thioredoxin (Trx), which is an antioxidant protein and plays pivotal roles in maintaining intracellular redox balance, inhibited ASK1 kinase activity by direct binding to ASK1 under normal conditions. Under oxidative conditions, ASK1 is dissociated from Trx and therefore fully activated. The active site of Trx contains two cysteine residues that undergo reversible oxidation to form a disulfide bond with each other, so that the conformation of Trx is changed by intracellular redox conditions. Thus, the oxidative stress-induced conformational change of Trx is particularly important for interaction with and regulation of ASK1, and elucidation of the regulatory mechanisms of ASK1 by Trx is critical to understanding the intracellular redox signaling. In this chapter, we review the regulatory mechanisms of ASK1 activity by Trx, and describe a method for monitoring in vitro binding between Trx and ASK1 under various redox conditions. In addition, we present methods to detect the oxidative stress-induced activation of ASK1 in the cells by Western blot analysis and in vitro kinase assay. The techniques presented in this chapter will be useful for a range of investigations into intracellular redox signaling.


Chromatin organization in the homogeneously staining regions of a methotrexate-resistant mouse cell line: interspersion of inactive and active chromatin domains distinguished by acetylation of histone H4.
We have analyzed the organization of the homogeneously staining regions (HSRs) in chromosomes from a methotrexate-resistant mouse melanoma cell line. Fluorescence in situ hybridization techniques were used to localize satellite DNA sequences and the amplified copies of the dihydrofolate reductase (DHFR) gene that confer drug-resistance, in combination with immunofluorescence using antibody probes to differentiate chromatin structure. We show that the major DNA species contained in the HSRs is mouse major satellite, confirming previous reports, and that this is interspersed with DHFR DNA in an alternating tandem array that can be resolved at the cytological level. Mouse minor satellite DNA, which is normally located at centromeres, is also distributed along the HSRs, but does not appear to interfere with centromere function. The blocks of major satellite DNA are coincident with chromatin domains that are labelled by an autoantibody that recognizes a mammalian homologue of Drosophila heterochromatin-associated protein 1, shown previously to be confined to centric heterochromatin in mouse. An antiserum that specifically recognizes acetylated histone H4, a marker for active chromatin, fails to bind to the satellite DNA domains, but labels the intervening segments containing DHFR DNA. We can find no evidence for the spreading of the inactive chromatin domains into adjacent active chromatin, even after extended passaging of cells in the absence of methotrexate selection.


The Wessex Fit-4-Cancer Surgery Trial (WesFit): a protocol for a factorial-design, pragmatic randomised-controlled trial investigating the effects of a multi-modal prehabilitation programme in patients undergoing elective major intra-cavity cancer surgery.
Background: Surgical resection remains the primary curative treatment for intra-cavity cancer. Low physical fitness and psychological factors such as depression are predictive of post-operative morbidity, mortality and length of hospital stay. Prolonged post-operative morbidity is associated with persistently elevated risk of premature death. We aim to investigate whether a structured, responsive exercise training programme, a psychological support programme or combined exercise and psychological support, delivered between treatment decision and major intra-cavity surgery for cancer, can reduce length of hospital stay, compared with standard care. Methods: WesFit is a pragmatic , 2x2 factorial-design, multi-centre, randomised-controlled trial, with planned recruitment of N=1560. Participants will be randomised to one of four groups. Group 1 (control) will receive usual pre-operative care, Group 2 (exercise) patients will undergo 2/3 aerobic, high-intensity interval training sessions per week supervised by personal trainers. Group 3 (psychological support) patients are offered 1 session per week at a local cancer support centre. Group 4 will receive both exercise and psychological support. All patients undergo baseline and pre-operative cardiopulmonary exercise testing, complete self-report questionnaires and will be followed up at 30 days, 12 weeks and 12 months post-operatively. Primary outcome is post-operative length-of-stay. Secondary outcomes include disability-adjusted survival at 1-year postoperatively, post-operative morbidity, and health-related quality of life. Exploratory investigations include objectively measured changes in physical fitness assessed by cardiopulmonary exercise test, disease-free and overall mortality at 1-year postoperatively, longer-term physical activity behaviour change, pre-operative radiological tumour regression, pathological tumour regression, pre and post-operative body composition analysis, health economics analysis and nutritional characterisation and its relationship to post-operative outcome. Conclusions: The WesFit trial will be the first randomised controlled study investigating whether an exercise training programme +/- psychological intervention results in improvements in clinical and patient reported outcomes in patients undergoing major inter-cavity resection of cancer. ClinicalTrials.gov registration: NCT03509428 (26/04/2018).


Hypothesis of a potential BrainBiota and its relation to CNS autoimmune inflammation.
Infectious agents have been long considered to play a role in the pathogenesis of neurological diseases as part of the interaction between genetic susceptibility and the environment. The role of bacteria in CNS autoimmunity has also been highlighted by changes in the diversity of gut microbiota in patients with neurological diseases such as Parkinson's disease, Alzheimer disease and multiple sclerosis, emphasizing the role of the gut-brain axis. We discuss the hypothesis of a brain microbiota, the BrainBiota: bacteria living in symbiosis with brain cells. Existence of various bacteria in the human brain is suggested by morphological evidence, presence of bacterial proteins, metabolites, transcripts and mucosal-associated invariant T cells. Based on our data, we discuss the hypothesis that these bacteria are an integral part of brain development and immune tolerance as well as directly linked to the gut microbiome. We further suggest that changes of the BrainBiota during brain diseases may be the consequence or cause of the chronic inflammation similarly to the gut microbiota.


Antiemetic effect of perphenazine versus prochlorperazine intravenously before cisplatin therapy.
Antiemetic effects of perphenazine and prochlorperazine, both administered by continuous i.v. infusion after a loading dose, were compared in patients receiving cisplatin. Study subjects were 6 men and 13 women for whom other antiemetic therapy had failed; each patient was studied during two courses of cisplatin therapy. Patients were randomly selected to receive either perphenazine or prochlorperazine during the first course; for the second course, each received the other antiemetic. During drug administration, nausea, retching, vomiting, and side effects of the antiemetic were recorded hourly by the patient and concurrently by a pharmacist observer (both blinded). Each patient's scores on nausea, retching, and vomiting were compared by drug and by treatment sequence. Evaluable data for 17 patients showed that aggregate differences between responses to the two drugs were not significant. Fourteen patients had significantly less nausea, retching, and vomiting during the second course of treatment. Few side effects were reported. Nervousness was experienced with prochlorperazine in four patients and perphenazine in one, and drowsiness occurred with prochlorperazine in four patients and perphenazine in three. Perphenazine and prochlorperazine, when given in equal doses and administered by continuous i.v. infusion after a loading dose, were equally effective in controlling nausea and vomiting associated with cisplatin therapy.