Management of breast cancer patients with chemotherapy-induced neutropenia or febrile neutropenia.
Chemotherapy-induced neutropenia (CIN) is a common toxicity caused by the administration of anticancer drugs. This side effect is associated with life-threatening infections and may alter the chemotherapy schedule, thus impacting on early and long-term outcomes. Elderly breast cancer patients with impaired health status or advanced disease as well as patients undergoing dose-dense anthracycline/taxane- or docetaxel-based regimens have the highest risk of CIN. A careful assessment of the baseline risk for CIN allows the selection of patients who need primary prophylaxis with granulocyte colony-stimulating factor (G-CSF) and/or antimicrobial agents. Neutropenic cancer patients may develop febrile neutropenia and CIN-related severe medical complications. Specific risk assessment scores, along with comprehensive clinical evaluation, are able to define a group of febrile patients with low risk for complications who can be safely treated as outpatients. Conversely, patients with higher risk of severe complications should be hospitalized and should receive intravenous antibiotic therapy with or without G-CSF.


Contribution of the Mission in Afghanistan to the Burden of Past-Year Mental Disorders in Canadian Armed Forces Personnel, 2013.
The purpose of this study was to estimate the contribution of the mission in Afghanistan to the burden of mental health problems in the Canadian Armed Forces (CAF). Data were obtained from the 2013 Canadian Forces Mental Health Survey, which assessed mental disorders using the World Health Organization's Composite International Diagnostic Interview. The sample consisted of 6696 Regular Force (RegF) personnel, 3384 of whom had deployed in support of the mission. We estimated the association of past-year mental health problems with Afghanistan deployment status, adjusting for covariates using logistic regression; population attributable fractions (PAFs) were also calculated. Indication of a past-year mental disorder was identified in 18.4% (95% confidence interval [CI], 17.0% to 19.7%) of Afghanistan deployers compared with 14.6% (95% CI, 13.3% to 15.8%) in others. Afghanistan-related deployments contributed to the burden of a past-year disorder (PAF = 8.7%; 95% CI, 3.0% to 14.2%), with the highest PAFs being seen for panic disorder (34.7%) and posttraumatic stress disorder (32.1%). The PAFs for individual alcohol use disorders and suicide ideation were not different from zero. Child abuse, however, had a much greater PAF for any past-year disorder (28.7%; 95% CI, 23.4% to 33.7%) than did the Afghanistan mission. The mission in Afghanistan contributed significantly to the burden of mental disorders in the CAF RegF in 2013. However, the much stronger contribution of child abuse highlights the need for strong military mental health systems, even in peacetime, and the need to target the full range of determinants of mental health in prevention and control efforts.


Lumping analysis of biochemical reaction systems with time scale separation.
Due to the complexity of the systems, successful modelling of intracellular reaction networks must rely on lumping techniques which systematically reduce the number of variables and parameters. Fortunately, the time scale separation characteristics of biochemical systems provide opportunities for eliminating unnecessary details. Through the proper interpretation of eigenvalues and eigenvectors, this article presents a theoretical basis for systematic model reduction. Results are generalized as a semiheuristic basis for lumping systems without complete kinetic information. It is also illustrated that the simplified system can yield new insight which is otherwise unavailable.


Competitive enzyme-liked immunoassay with use of soluble enzyme/antibody immune complexes for labeling. I. Measurement of human choriogonadotropin.
We described the principle of a new enzyme-immunoassay, competitive enzyme-liked immunoassay (CELIA), for quantitative measurement of soluble antigens and haptens. In the assay, binding of antibody to antigen-immunosorbent is competitively inhibited by the free antigen to be measured. The amount of first antibody bound to the immunosorbent is measured by an enzymatic technique in which a heterologous bridging antibody and a soluble antibody/enzyme immune complex are applied in sequence. The soluble complex we used was rabbit antiperoxidase/horseradish peroxidase. Peroxidase activity is inversely proportional to the concentration in the original sample of the substance to be assayed. The enzyme-linked reagents are potentially widely applicable to any substance to be measured. To demonstrate the feasibility of CELIA, we report a preliminary study of its application to the measurement of human chloriogonadotropin in serum and urine. The assay described for this hormone has a working range of 1 to 50 int. units per milliliter of sample. The technique obviates the disadvantages associated with measurement and handling of radioisotopes in radioimmunoassays and the only major instrumentation required is a centrifuge and a conventional spectrophotometer.


Pathologic Response of HER2-positive Gastric Cancer to Trastuzumab-based Chemotherapy.
Trastuzumab-based chemotherapy is now a standard approach for patients with human epidermal growth factor receptor 2 (HER2)-positive advanced gastric cancer. However, histopathologic changes after treatment have yet to be elucidated. This study aims to characterize the histologic response of gastric cancer to trastuzumab treatment and its correlation with HER2 status. Twenty-one advanced HER2-positive gastric cancers treated with trastuzumab-based chemotherapy, including 10 surgically resected specimens and 11 biopsy samples from patients with inoperable tumors, were evaluated for the histologic responses and HER2 status of residual cells. We also reviewed, as controls, 10 cases undergoing surgical resection of tumors after chemotherapy without trastuzumab. Complete and partial histologic responses were obtained in 2 and 8 of the surgical cases, respectively. HER2-positive neoplastic cells were recognized at least focally in the 8 cases. Notably, the proportion of HER2-positive cells was always higher in superficial (mucosal/submucosal) layers than in deeper layers. Three specimens contained HER2-positive neoplastic cells exclusively in the superficial area or intravascular space, whereas deeply invasive or metastatic components almost completely disappeared or were HER2 negative when still present. In contrast, HER2-negative cells or residual tumor cells in control cases tended to survive as well or better in deeply invasive areas or in metastases than in superficial areas. Biopsy samples from nonoperative patients remained HER2 positive after treatment in 8 of 11 patients. Our observations suggest that HER2-positive neoplastic cells tend to survive within superficial areas or intravascular spaces after trastuzumab therapy, even when deeply invasive or metastatic lesions responded well to therapy.


Skeletal muscle metabolism as a target for drug therapy in peripheral arterial disease.
Peripheral arterial disease (PAD) is an atherosclerotic disease which modifies lower extremity hemodynamics. There is considerable evidence that skeletal muscle metabolism is altered in PAD. Several studies have demonstrated altered mitochondrial enzyme content in PAD muscle as compared with controls, and enzyme activity may not increase normally in PAD with exercise training. A variety of metabolic intermediates, including acylcarnitines, accumulate in muscle of PAD patients, suggesting incomplete oxidative metabolism. Studies employing 31P-NMR (nuclear magnetic resonance) also suggest a metabolic myopathy in PAD. Strikingly, while hemodynamics do not predict claudication-limited performance, metabolic injury as evidenced by acylcarnitine accumulation is strongly correlated with patients' functional status in PAD. Further, exercise rehabilitation improves claudication-limited performance without modifying large vessel hemodynamics. The stress placed on skeletal muscle during exercise in PAD and the observed evidence of metabolic dysfunction is similar to ischemia/reperfusion injury in cardiac muscle. Recognition of the role of cellular metabolic injury and function in PAD has formed the basis for novel therapeutic strategies in this disease.


COX-2 expression in highly aggressive thyroid malignancies - indication for a possible therapeutic option?
Both anaplastic thyroid carcinoma (ATC) and angiosarcoma of the thyroid (AST) are highly aggressive malignancies with very limited therapeutic options. Since selective inhibition of COX-2, for example, by celecoxib has been shown to suppress both tumour formation and progression, we investigated COX-2 protein expression in a series of ATC and AST (26 cases each) using immunohistochemistry. COX-2 expression was demonstrated in 13 ATC (50%) and 11 AST (42%); a strong COX-2 expression in more than 50% of vital tumour cells was found in 5 ATC and 5 AST, respectively. Although a recently performed phase II trial applying celecoxib failed overall to halt tumour progression in differentiated thyroid carcinoma, the two cases with partial or complete remission noted in this study were related to tumours with immunohistochemically proven strong COX-2 expression. The strong COX-2 expression observed in approximately 20% of our ATC and AST samples may thus indicate selective patients with a possible therapeutic option for an otherwise fatal disease.


Hospital personnel in surgical services in a transplant hospital faced with living liver donation: an attitudinal survey.
Living liver donation is becoming a more widely accepted practice given the decrease in donor morbidity and mortality and the beneficial results in the recipient. Use of this technique is unusual in Spain. There are a number of workers against its use within the health care system. The objective of this study was to analyze attitudes toward and the variables that affect them concerning living liver donation among surgical services in a transplant hospital. A random survey was stratified by surgical area and job category in a Spanish third-level hospital with an ongoing solid organ transplant program. Attitudes toward living liver donation were assessed using a questionnaire about organ donation and transplantation that evaluated various psychosocial variables. Student's t test and the chi square test were used. A total of 263 workers were surveyed (mean age as 40 +/- 9 years). Regarding attitudes toward living liver donation, the level of acceptance was 80% (n = 211) of respondents, whereas 10% were undecided (n = 26), and another 10% were against (n = 26), assuming that the donations were related. When we asked about unrelated living donation, the percentage in favor decreased to 10% (n = 27). No differences were found with respect to job category or type of service. The variables that are related to such an attitude are the following: (1) possibility of respondent needing an organ (P = .001); (2) favorable attitude toward living kidney donation (P < .000); and (3) a belief that medical errors exist (P = .004). An important finding was that attitudes toward living liver donation were not more favorable according to whether the respondent was in favor or against cadaveric organ donation (P = .175). There was a highly favorable attitude toward living liver donation among hospital personnel in surgical services, which is an important factor to take into account if this type of donation is to be encouraged in Spain.


Increased serum IgA in Fcα/μR-deficient mice on the (129 x C57BL/6) F1 genetic background.
Fcα/μR (CD351) is an Fc receptor for both IgA and IgM, which is abundantly expressed in the small intestine. However, the role of Fcα/μR in the intestinal tissue is largely unknown. Here, we found that Fcα/μR is highly expressed on follicular dendritic cells (FDCs) in Peyer's patches (PP) in the small intestine. Fcα/μR-deficient mice on the (129 x C57BL/6) F1 background showed increased serum, but not fecal, IgA level in response to gut-oriented antigens. IgA(+) B cells were increased in PP, but not in the lamina propria, of Fcα/μR-deficient mice, which was attenuated after reduction of commensal microbiota by oral treatment with antibiotics. Analyses of bone marrow chimeric mice, in which either FDCs or blood cells or both lack the expression of Fcα/μR, suggested that FDCs, but not blood cells, were responsible for the increased serum IgA concentration in Fcα/μR-deficient mice. Moreover, Fcα/μR-deficient mice showed enhanced germinal center formation against commensal microbiota in PP. Thus, serum IgA production against gut-oriented antigens is negatively regulated by Fcα/μR on FDCs in the F1 mice.


Statin-induced Ras activation integrates the phosphatidylinositol 3-kinase signal to Akt and MAPK for bone morphogenetic protein-2 expression in osteoblast differentiation.
Lovastatin promotes osteoblast differentiation by increasing bone morphogenetic protein-2 (BMP-2) expression. We demonstrate that lovastatin stimulates tyrosine phosphorylation of the p85 regulatory subunit of phosphatidylinositol 3-kinase (PI3K), leading to an increase in its kinase activity in osteoblast cells. Inhibition of PI3K ameliorated expression of the osteogenic markers alkaline phosphatase, type I collagen, osteopontin, and BMP-2. Expression of dominant-negative PI3K and PTEN, an inhibitor of PI3K signaling, significantly attenuated lovastatin-induced transcription of BMP-2. Akt kinase was also activated in a PI3K-dependent manner. However, our data suggest involvement of an additional signaling pathway. Lovastatin-induced Erk1/2 activity contributed to BMP-2 transcription. Inhibition of PI3K abrogated Erk1/2 activity in response to lovastatin, indicating the presence of a signal relay between them. We provide, as a mechanism of this cross-talk, the first evidence that lovastatin stimulates rapid activation of Ras, which associates with and activates PI3K in the plasma membrane, which in turn regulates Akt and Erk1/2 to induce BMP-2 expression for osteoblast differentiation.


Acute Appendicitis: Epidemiological, Clinical, Surgical, and Post-surgical Characteristics in a Honduran General Hospital.
Acute appendicitis is a common surgical emergency worldwide, yet data specific to Central America, including Honduras, are limited. This study aimed to investigate the epidemiological, clinical, surgical, and post-surgical characteristics of acute appendicitis in a Honduran general hospital. A descriptive, quantitative, non-experimental, cross-sectional study was conducted at the Mario Catarino Rivas Hospital in San Pedro Sula, Honduras. The study sample consisted of 100 patients admitted with acute appendicitis from January to April 2022. Data on demographic factors, surgical interventions, appendicitis phases, appendix location, and laboratory findings were collected and analyzed. The mean age of the participants was 28.5 years, with a slight male predominance (52%). Timely surgical intervention was performed in 95% of cases within the first 12 hours. The gangrenous phase was observed in 30% of patients, followed by the perforated (24%), edematous (24%), and suppurative (22%). Retrocecal appendicitis accounted for the majority of cases (66%). Moderate leucocytosis (46%) and severe leucocytosis (39%) were associated with acute appendicitis severity. A higher neutrophil percentage was indicative of complicated appendicitis. Computed tomography was underutilized, with only one patient undergoing the examination. This study provides valuable insights into Honduras' epidemiological, clinical, and surgical characteristics of acute appendicitis. Early surgical intervention and laboratory findings, such as leukocyte count and neutrophil percentage, can aid in assessing disease severity. Further research is warranted to understand the unique aspects of acute appendicitis in Central America and optimize patient management. This study highlights the need for multi-centre studies and long-term follow-up to enhance our understanding of appendicitis in similar populations.


Effect of high-frequency oral airway and chest wall oscillation and conventional chest physical therapy on expectoration in patients with stable cystic fibrosis.
To compare the effect of high-frequency oral airway oscillation, high-frequency chest wall oscillation, and conventional chest physical therapy (CPT) on weight of expectorated sputum, pulmonary function, and oxygen saturation in outpatients with stable cystic fibrosis (CF). Prospective randomized trial. Pediatric pulmonary division of a tertiary care center. Fourteen outpatients with stable CF recruited from the CF center. Two modes of oral airway oscillation (1: frequency 8 Hz; inspiratory to expiratory [I:E] ratio 9:1; 2: frequency 14 Hz; I:E ratio 8:1), two modes of chest wall oscillation (1: frequency 3 Hz; I:E ratio 4:1; 2: frequency 16 Hz; I:E ratio 1:1, alternating with frequency 1.5 Hz, I:E ratio 6:1), and CPT (clapping, vibration, postural drainage, and encouraged coughing) were applied during the first 20 min of 4 consecutive hours. Sputum was collected on an hourly basis for a total of 6 consecutive hours. During the first and the last hour, patients collected sputum without having any treatment and underwent pulmonary function tests (PFTs). Oxygen saturation was measured at 30-min intervals during hours 1 to 6. For the first 20 min of the second to the fifth hour, patients received one of the treatments. To assess the effect of the intervention, the weight of expectorated sputum during hours 2 to 6 was averaged and expressed as percentage of the weight expectorated during the first hour (baseline). For the five treatment modalities, mean sputum dry and wet weights ranged between 122% and 185% of baseline. There was no statistically significant difference among the treatment modalities. As measured by sputum wet weight, all oscillatory devices tended to be less effective than CPT (p=0.15). As measured by dry weight, oral airway oscillation at 8 Hz with an I:E ratio of 9:1 and CPT tended to be more effective than the other treatment modalities (p=0.57). None of the treatment modalities had an effect on PFTs and oxygen saturation and all were well tolerated. In outpatients with stable CF, high-frequency oscillation applied via the airway opening or via the chest wall and CPT have comparable augmenting effects on expectorated sputum weight without changing PFTs or oxygen saturation. In contrast to CPT, high-frequency oral airway and chest wall oscillations are self-administered, thereby containing health-care expenses.


[Wegener's granulomatosis--diagnostic problem].
Wegener's granulomatosis there is a small--and middle--vessels vasculitis. The pathomorphological diagnostic criteria is known as Wegener's triad: 1) necrotizing granlomatous inflammation of upper and/or lower respiratory tract, 2) systemic or focal necrotizing vasculitis involving arteries and vein, and 3) focal segmental necrotizing crescentic gromerulonephritis. According to the current theory of pathogenesis of Wegener's granulomatosis, Staphylococcus aureus is involved. The rise in ANCA level during vascular inflammation is very important for monitoring disease. The main localization of Wagener's granulomatosis is in lung, nasal sinusites, nose, kidneys and nasal pharynx. The history of 60-years old female patient who was being treated in Department of Otolaryngology for chronic bilateral otitis media for many years is presented. During hospitalization the sinusites, rhinitis and renal failure additionally were found. The CT sinuses and head, histopatological and ANCA exam in patient were performed. Based on clinical, physical, additional exam Wegener's granulomatosis was diagnosed. 1) Surgical procedure is not indicated method of treatment of Wegener's granulomatosis, because it can increase the pathologic process. 2) The prognosis of Wegener's granulomatosis depends mainly on the ability to diagnose the early disease and the aplication of adequate treatment.


Developmental changes in extracellular matrix messenger RNAs in the mouse placenta during the second half of pregnancy: possible factors involved in the regulation of placental extracellular matrix expression.
Expression of procollagens (Col1a1/2, Col3a1, Col4a1/2, Col5a1/2) and fibronectin 1 (Fn1) in the mouse fetal placental tissue was examined during the second half of pregnancy. Ribonuclease protection assays (RPAs) revealed that levels of these mRNAs noticeably increased between Days 10 and 14 of pregnancy, and they remained at relatively constant levels thereafter. In situ hyridization showed that Col1a1 and Col4a1 mainly localized in the labyrinth, whereas Fn1 was expressed mainly in the spongiotrophoblast. Since members of the transforming growth factor-beta (TGFB) superfamily are involved in the regulation of extracellular matrix (ECM) expression in various tissues, mRNA levels of TGFB family members and their binding proteins were also examined by RPAs. Transforming growth factor-beta1-3 (Tgfb1-3), activin subunits (Inhba, Inhbb), follistatin (Fst), and follistatin-like 3 (Fstl3) were expressed in the placenta, whereas significant expression of myostatin (Mstn) was not detected. Although the expression patterns of Tgfb1-3 and Inhba in the placenta suggest possible involvement of TGFBs and activin A in the regulation of placental ECM expression, neither TGFBs nor activin A affected ECM mRNA levels in vitro. On the other hand, hypoxia significantly decreased Col1a1/2 and Col4a1/2 mRNAs in cultured placental cells, and a high-glucose condition significantly increased Col1a1 and Col3a1 mRNAs. Fn1 expression was increased under the high-glucose condition, although hypoxia also increased Fn1 expression to a lesser degree. These data suggest that an increase in oxygen tension and nutrient supply during placentation rather than TGFB family members may be responsible for the increase in the placental ECM mRNA expression.


Four novel GALC gene mutations in two Chinese patients with Krabbe disease.
Krabbe disease (OMIM #245200) is a rare autosomal recessive leukodystrophy caused by deficiency of galactocerebrosidase (GALC) activity. We identified four novel mutations of the GALC gene in two unrelated Chinese families with Krabbe disease: one insertion mutation, c.1836_1837insT, and one nonsense mutation, c.599C>A (p.S200X), in an infantile patient, and one deletion mutation, c.1911+1_1911+5delGTAAG, and one missense mutation, c.2041G>A, in an adult late-onset patient. This is the first identification of GALC mutations in the Chinese population.


miR-17-5p/20a are important markers for gastric cancer and murine double minute 2 participates in their functional regulation.
To investigate the potential roles and mechanisms of miR-17-5p/20a in human gastric cancer development and progression. Quantitative real-time polymerase chain reaction (qRT-PCR) was employed to determine miR-17-5p/20a expression profiles in 110 gastric cancer tissues. microRNAs' (miRNAs) mimics and inhibitors were used to reveal their function in gastric cancer. Antagomirs were applied to treating gastric cancer cell derived xenograft in vivo. Western blot and luciferase assays were performed to uncover the targets and mechanisms of miR-17-5p/20a. miR-17-5p/20a levels were upregulated in human gastric cancer tissues. Overexpression of miR-17-5p/20a promoted gastric cancer cell cycle progression and inhibited cell apoptosis, whereas knockdown of miR-17-5p/20a resulted in cell cycle arrest and increased apoptosis. p21 and tumour protein p53-induced nuclear protein 1 (TP53INP1) were validated as the targets of miR-17-5p/20a. Antagomirs against miR-17-5p/20a significantly inhibited gastric cancer growth via upregulation of p21 and TP53INP1 in a mouse xenograft model. The negative relationship between miR-17-5p/20a and TP53INP1 was observed in patient gastric cancer tissues. Murine double minute 2 (MDM2) was found to be involved in miRNA regulation and function. Targeted inhibition of MDM2 in a miRNA mimic-transfected gastric cancer cell line abolished miR-17-5p/20a function and inhibition of p21 expression. MDM2 restoration by pCMV-MDM2 rescued the functionality. Our findings indicate that miR-17-5p/20a promote gastric cancer cell proliferation and inhibit cell apoptosis via post-transcriptional modulation of p21 and TP53INP1. They may be promising therapeutic markers for gastric cancer. MDM2 contributes to miR-17-5p/20a function and inhibition of p21 in gastric cancer, and may be a novel mechanism underlying the oncogenic roles of miR-17-5p/20a.


Association of energy intake and expenditure with obesity: A cross-sectional study of 150 pediatric patients following treatment for leukemia.
Increased obesity in leukemia survivors has been attributed to chemotherapy and radiation. Data on total energy intake (TEI) and total energy expenditure (TEE) are lacking in obese childhood leukemia patients after completion of therapy from India. We conducted a cross-sectional study in pediatric acute leukemia patients after completion of therapy wherein energy intake was assessed by 24-hour recall method. TEE was calculated using Harris-Benedict equation, by assessing the physical activity level using Physical Activity Questionnaire for children and basal metabolic rate by World Health Organization equation. Indian Academy of Pediatrics 2015 guidelines for BMI were used for defining overweight and obesity. Nutritional status was assessed in 150 leukemia patients after completion of therapy. Twenty-five percent of leukemia patients after completion of therapy were overweight and obese versus 11% of healthy controls (p = 0.042). The mean ratio of TEI/required energy intake (REI), TEE/required energy expenditure (REE), and (TEI:REI)/(TEE:REE) were significantly higher in overweight and obese group versus nonobese survivors (p < 0.001, p = 0.091, p < 0.001, respectively). Multivariate analysis showed higher income (HR-2.3, p = 0.04), increased TEI/REI (HR-4, p = 0.049) and higher (TEI:REI)/(TEE:REE) (HR-3.1, p = 0.039) to be significant factors predicting obesity. Obesity in leukemia patients after completion of therapy is associated with increased energy intake, causing imbalance between energy intake and TEE in these patients.


A Simple, Cleated DNA Walker That Hangs on to Surfaces.
We designed and demonstrated a single-legged or unipedal walker that has a "cleat" that allows it to persistently associate with a track and make autonomous decisions about movement. The walker is highly processive over long periods of time, as shown by its movement over a microparticle surface suffused with substrate. The simple design can be readily optimized on the basis of simple energetic considerations. The walker can be used for signal amplification and should prove especially valuable for programming amorphous computations within chemical reaction networks.


Chronic thromboembolic pulmonary hypertension and vascular transformation of the lymph node sinuses.
Chronic thromboembolic pulmonary hypertension (CTEPH) is a disease often not recognized until it is far advanced. Medical management does not reverse the effects of the disease, nor does it prevent its progression. Pulmonary thromboendarterectomy is the preferred treatment. We present the case of a patient in whom CTEPH was mistaken for an interstitial lung disease with mediastinal lymph node enlargement. The mediastinal lymph node enlargement was due to vascular transformation of the lymph node sinuses (VTS). This is an unusual case of chronic thromboembolic pulmonary hypertension with vascular transformation of the lymph node sinuses.


Efficient and stable CRISPR/Cas9-mediated genome-editing of human type 2 innate lymphoid cells.
Innate lymphoid cells (ILCs) are a family of innate lymphocytes with important roles in immune response coordination and maintenance of tissue homeostasis. The ILC family includes group 1 (ILC1s), group 2 (ILC2s) and group 3 (ILC3s) 'helper' ILCs, as well as cytotoxic Natural Killer (NK) cells. Study of helper ILCs in humans presents several challenges, including their low proportions in peripheral blood or needing access to rare samples to study tissue resident ILC populations. In addition, the lack of established protocols harnessing genetic manipulation platforms has limited the ability to explore molecular mechanism regulating human helper ILC biology. CRISPR/Cas9 is an efficient genome editing tool that enables the knockout of genes of interest, and is commonly used to study molecular regulation of many immune cell types. Here, we developed methods to efficiently knockout genes of interest in human ILC2s. We discuss challenges and lessons learned from our CRISPR/Cas9 gene editing optimizations using a nucleofection transfection approach and test a range of conditions and nucleofection settings to obtain a protocol that achieves effective and stable gene knockout while maintaining optimal cell viability. Using IL-4 as a representative target, we compare different ribonucleoprotein configurations, as well as assess effects of length of time in culture and other parameters that impact CRISPR/Cas9 transfection efficiency. Collectively, we detail a CRISPR/Cas9 protocol for efficient genetic knockout to aid in studying molecular mechanism regulating human ILC2s.


Electromyographic biofeedback for gait training after stroke.
To examine the effects of electromyographic (EMG) biofeedback training on the recovery of gait in the acute phase post stroke. Patients were randomly assigned to EMG biofeedback or control groups. They received treatment three times a week for six weeks. All patients were assessed prior to treatment, after 18 treatment sessions, and at three months follow-up. The study was carried out at Scunthorpe General Hospital in North Lincolnshire. The subjects were acute stroke patients who had been admitted on to the medical and elderly wards. The EMG biofeedback group were treated using EMG as an adjunct to physiotherapy. The patients were encouraged to facilitate or inhibit abnormal muscle tone via auditory or visual signals transmitted from electrodes placed over the appropriate muscles. The control group were treated using the same techniques, electrodes were used with this group of patients, but the EMG machine was turned off and faced away from the patient and the therapist to control the placebo effect. A large battery of outcome measures was used for physical and psychological assessment. The physical measures consisted of active movement, muscle tone, sensation, proprioception, mobility and activities of daily living (ADL). The psychological measures included orientation, memory, spatial performance, language and IQ. Twenty-one patients were included in the study. Scores were combined into four groups: mild EMG, severe EMG, mild control and severe control. Results showed that there was an improvement in physical scores for active movement, mobility and ADL over time, but there was no significant difference between the EMG and control groups. Scores on the psychological tests were within normal limits, and there was no difference in performance between the EMG and control groups. This study showed no significant differences in the rate of improvement after stroke between the two groups. Although EMG biofeedback was used as an adjunct to physiotherapy and represented clinical practice, the results provide little evidence to support the clinical significance of using EMG biofeedback to improve gait in the acute phase after stroke.


Brazilian guidelines on prevention of cardiovascular disease in patients with diabetes: a position statement from the Brazilian Diabetes Society (SBD), the Brazilian Cardiology Society (SBC) and the Brazilian Endocrinology and Metabolism Society (SBEM).
Since the first position statement on diabetes and cardiovascular prevention published in 2014 by the Brazilian Diabetes Society, the current view on primary and secondary prevention in diabetes has evolved as a result of new approaches on cardiovascular risk stratification, new cholesterol lowering drugs, and new anti-hyperglycemic drugs. Importantly, a pattern of risk heterogeneity has emerged, showing that not all diabetic patients are at high or very high risk. In fact, most younger patients who have no overt cardiovascular risk factors may be more adequately classified as being at intermediate or even low cardiovascular risk. Thus, there is a need for cardiovascular risk stratification in patients with diabetes. The present panel reviews the best current evidence and proposes a practical risk-based approach on treatment for patients with diabetes. The Brazilian Diabetes Society, the Brazilian Society of Cardiology, and the Brazilian Endocrinology and Metabolism Society gathered to form an expert panel including 28 cardiologists and endocrinologists to review the best available evidence and to draft up-to-date an evidence-based guideline with practical recommendations for risk stratification and prevention of cardiovascular disease in diabetes. The guideline includes 59 recommendations covering: (1) the impact of new anti-hyperglycemic drugs and new lipid lowering drugs on cardiovascular risk; (2) a guide to statin use, including new definitions of LDL-cholesterol and in non-HDL-cholesterol targets; (3) evaluation of silent myocardial ischemia and subclinical atherosclerosis in patients with diabetes; (4) hypertension treatment; and (5) the use of antiplatelet therapy. Diabetes is a heterogeneous disease. Although cardiovascular risk is increased in most patients, those without risk factors or evidence of sub-clinical atherosclerosis are at a lower risk. Optimal management must rely on an approach that will cover both cardiovascular disease prevention in individuals in the highest risk as well as protection from overtreatment in those at lower risk. Thus, cardiovascular prevention strategies should be individualized according to cardiovascular risk while intensification of treatment should focus on those at higher risk.


Helicobacter pylori CagA and gastric carcinogenesis.
This study aimed to demonstrate the tyrosine phosphorylation motif (TPM) and 3' region structure of the Helicobacter pylori CagA gene as well as its SHP-2 binding activity in AGS cells and relation to gastric carcinogenesis. Sixteen clinical isolate H. pylori strains from eight duodenal ulcer and eight gastric adenocarcinoma patients were studied for CagA repeat sequence EPIYA motifs, C-terminal structure, and western blot analysis of CagA protein expression, translocation, and SHP-2 binding in AGS cells. Except for strain 547, all strains from the gastric adenocarcinoma patients were positive for CagA by PCR and had three EPIYA copy motifs. Western blotting showed that all strains were positive for CagA protein expression (100%), CagA protein translocation (100%), and SHP-2 binding (100%). CagA protein expression was significantly higher in the gastric adenocarcinoma patients than in the duodenal ulcer patients (P=0.0023). CagA protein translocation and SHP-2 binding in the gastric adenocarcinoma patients were higher than those in the duodenal ulcer patients, but no significant differences were found between the two groups (P=0.59, P=0.21, respectively). The TPMs and 3' region structures of the H. pylori CagA gene in the duodenal ulcer and gastric adenocarcinoma patients have no significant differences.


Studies on cation-induced membrane vesicle aggregation of porcine intestinal brush borders.
Cation specificity and effects of pH and temperature on membrane aggregation of porcine intestinal brush borders were studied in terms of the turbidity change of the membrane suspensions due to the addition of polyvalent cations. Monovalent cations (K+ and Na+) did not induce the membrane aggregation at 5 mM or below. Divalent and trivalent cations induced membrane aggregation even below 0.5 mM in the following orders of effectiveness: Hg2+ much greater than Cd2+ greater than Zn2+ greater than Mn2+ greater than Ca2+ greater than or equal to Ba2+ greater than Sr2+ greater than or equal to Mg2+ and Lu3+ greater than Yb3+ greater than Nd3+ = Er3+ = Tb3+ greater than Ce3+ greater than La3+, respectively. The aggregation of membranes induced by these polyvalent cations was reversible. Membrane aggregation induced by either divalent or trivalent cations followed saturation kinetics with increasing cation concentration and the apparent dissociation constants of these cations for the membranes were estimated to be at 4-6 mM for divalent cations (Ca2+, Mn2+, and Sr2+) and 1-2 mM for trivalent cations (Lu3+, Tb3+, and La3+). Cd2+ and Hg2+ had peculiarly much lower dissociation constants than any other divalent cations tested, being rather comparable to trivalent ones. The membrane aggregation induced by Ca2+, Mn2+, or Sr2+ was independent of pH in the range of 6.6 to 8.0, whereas that induced by Tb3+ or Cd2+ was linearly enhanced with increasing pH, very steeply above pH 7.0. The membrane aggregation induced by Cd2+, Hg2+, or Tb3+ was enhanced steadily with increasing temperature between 15 and 45 degrees C. On the other hand, Ca2+-induced aggregation showed a peak at around 30 degrees C in the temperature dependence profile, still being enhanced above 40 degrees C.


Relationship between health service use and health information technology use among older adults: analysis of the US National Health Interview Survey.
Older adults are the most frequent and heaviest users of health services in the United States; however, previous research on older adults' use of health information technology (HIT) has not examined the possible association of HIT use among older adults with their use of health services. This study examined the relationship between US older adults' use of health services and their use of the Internet for health-related activities, controlling for socioeconomic characteristics and aging-related limitations in sensory and cognitive function. It also examined gender differences in the pattern of association between the types of health services used and HIT use. The data for this study were drawn from the 2009 US National Health Interview Survey (NHIS), which was the first nationally representative household survey to collect data on HIT (Internet) use. First, the rates of lifetime and 12-month HIT use among sample adults (n = 27,731) by age group (18-29 to 85 and over) were analyzed. Second, bivariate analysis of sociodemographic characteristics, health status, and health service use by HIT use status among those aged 65 or older (n = 5294) was conducted. Finally, multivariate binary logistic regression analysis was used to test the study hypotheses with 12-month HIT use as the dependent variable and 12-month health service uses among the age group 65 or older as possible correlates. The rates of HIT use were significantly lower among the age groups 65 or older compared with the younger age groups, although the age group 55 to 64 was not different from those younger. The rates of HIT use decreased from 32.2% in the age group 65 to 74 to 14.5% in the age group 75 to 84 and 4.9% in the 85 and older age group. For both genders, having seen or talked to a general practitioner increased the odds of HIT use. However, having seen or talked to a medical specialist, eye doctor, or physical therapist/occupational therapist (PT/OT) were significantly associated with HIT use only for older women, while having seen or talked to a mental health professional only marginally increased the odds of HIT use only for older men. Having visited or talked to a chiropractor and having had overnight hospitalization, surgery, and/or homecare services were not associated with the odds of HIT use for either gender. Older-adult users of general health services were more likely to use HIT than nonusers of general health services, while older-adult users of specialized health services were not different from nonusers of specialized health services in their odds of HIT use. The findings have implications for narrowing the age-related and socioeconomic status-related gaps in HIT use. The access gaps among racial/ethnic minority older adults and poorly educated and/or low-income older adults are especially striking and call for concerted efforts to facilitate Internet access and HIT use among these disadvantaged older adults.


Spatio-temporal expression of chromogranin A during zebrafish embryogenesis.
Chromogranin A (CHGA), a protein participating in the biogenesis of dense core secretory granules in various neuroendocrine tissues, plays a critical role in the release of hormones/peptides and the pathogenesis of pheochromocytoma. However, little is known about the developmental origin of CHGA-expressing cells during embryogenesis. Here, we report the structural characterization and spatio-temporal expression pattern of zebrafish (Danio rerio) ortholog of mammalian CHGA. The earliest expression of chga transcripts was observed at 16 h post fertilization in the developing cranial ganglia as six distinct cellular masses arranged bilaterally as strings of beads in the dorsal root ganglia (DRG) precursors along the dorsal trunk. With development advancing, the chga transcripts were expressed abundantly in diencephalon, mesencephalon, and rhombencephalon as well as in the DRG. Interestingly, double in situ hybridization assay of chga with genes expressed in pronephros (Wilms' tumor suppressor 1, wt1), adrenal cortex (side-chain cleavage enzyme, scc), and sympathoadrenal neuron/chromaffin cell (dopamine-beta-hydroxylase, dbh), respectively, showed that the chga-expressing cells are spatially separated from wt1-, scc-, and dbh-positive cell populations during early embryonic development. The pronephros region does not express chga even up to 7 days post fertilization, while chga positive-staining cells bind in the brain and DRG, indicating that chga may play an important role in nervous system development during the early embryonic stages.


Discovery and characterization of the tuberculosis drug lead ecumicin.
The new tuberculosis (TB) lead ecumicin (1), a cyclic tridecapeptide, was isolated from Nonomuraea sp. MJM5123, following a high-throughput campaign for anti-TB activity. The large molecular weight of 1599 amu detected by LC-HR-MS precluded the initial inference of its molecular formula. The individual building blocks were identified by extensive NMR experiments. The resulting two possible planar structures were distinguished by LC-MS(2). Determination of absolute configuration and unambiguous structural confirmation were carried out by X-ray crystallography and Marfey's analysis.


ASSOCIATION BETWEEN SHORT STATURE AND HAIR ELEMENTS.
Aim - assessment of hair elemental status, determination of elemental imbalances and heavy metal concentration and evaluation its impact on child liner growth. Case-control study involves 112 children less than 5 years old (target group - 54 children with short stature, control - 58 children with normal physical development). Medical history, nutritional status, anthropological parameters were assessed. Patients with genetic and endocrine disorders, family short stature were excluded from the study. Child elemental status was detected in the hair, with roentgen-fluorescence spectrometer method. Statistical analysis was conduced using SPPS19. Assessment of the dietary history does not reveal any significant differences between the groups, the only exception was consumption of fish, that was statistically significantly lower than in control (p<0,05). The study revealed deficiency of some elements in both (study and control) groups, but there were significant difference between the groups: Zn deficiency 90% versus 40% (p<0,05), Ca deficiency 62 and 36 (p<0,05), Cu deficiency 50 and 16 (p<0,05) and Mn deficiency 30 and 6 (p<0,05) accordingly. Level of some elements (K, S, Br, Cl, Co, Ag, V, Ni, Rb, Sr, Ti, Ba, As, Cd, Zr, Sb) have normal values. Some element deficiency (Fe, Mo, Se) was higher in study group but the difference was not significant. The study revealed high level of hair lead in both groups: all patients (100%) of target group have elevated level of hair lead, among them in 35,8% the content of lead was above so-called minimal allowed level (p<5,0 ppm) and in 64,5% the content of lead in the hair was above so-called maximal allowed level (p≥5,0 ppm). In the control group, lead in the hair was detected in 78% of cases, though the concentration was only in 2% of cases higher than so called maximum allowed. Our study clearly indicated deficiency of trace elements, particularly essential ones and high lead contamination in children with short stature. Our study proves, that analyzing of trace elements and heavy metals levels and evaluation of its influence on health and development have great importance especially during early years, in the critical period of active growth and development.


Risk factors for anorexia nervosa: a national cohort study.
To identify possible risk factors for anorexia nervosa through national registers. The study includes the entire Swedish population of 989,871 individuals born between 1973 and 1982. Patients with anorexia nervosa were identified through the Swedish Hospital Discharge Register from November 1990 to December 1999. Information about sociodemographic, perinatal, and psychosocial variables was obtained from different national registers. Attributable risk (AR) was calculated for potential risk factors. Females had the highest AR for hospital admission because of anorexia nervosa (89.2%). Another important risk factor was having parents born in northern, central, or eastern Europe (AR: 49.3%). Psychosocial risk factors also were associated with an increased risk for anorexia nervosa (AR: 7.6%), whereas perinatal complications had an AR of only 3.6%. The most important risk factors were related to the sociocultural context of the individual, thus supporting hypotheses of a sociocultural etiology of anorexia nervosa.


Endothelin-1-induced myocardial ischaemia and oedema in the rat: involvement of the ETA receptor, platelet-activating factor and thromboxane A2.
1. The objectives of the present experiments were to assess the role of ETA receptors in mediating endothelin-1 (ET-1)-induced myocardial ischaemia and oedema and to study the involvement of platelet-activating factor (PAF) and thromboxane A2 (TxA2) in these actions of ET-1 in rats. 2. Intravenous bolus injection of ET-1 (0.1-2 nmol kg-1) into anaesthetized rats induced ST segment elevation of the electrocardiogram in a dose-dependent manner without causing arrhythmias. ST segment elevation developed within 20-90 s and persisted for at least 10-20 min following administration of ET-1. 3. Pretreatment of the animals with the selective endothelin ETA receptor antagonist, FR 139317 (2.5 mg kg-1, i.v.) inhibited by 86% the ST segment elevation elicited by ET-1 (1 nmol kg-1). Pretreatment with intravenous administration of BM 13505 (1 mg kg-1), a TxA2 receptor antagonist, OKY-046 (10 mg kg-1), a thromboxane synthase inhibitor or the specific PAF receptor antagonist, WEB 2086 (1 mg kg-1) or BN 52021 (10 mg kg-1) markedly suppressed ST segment elevation in response to ET-1. Infusion of indomethacin (3 mg kg-1 bolus plus 2 mg kg-1 h-1) did not significantly affect ET-1-induced ST segment elevation. 4. Bolus injection of ET-1 (1 nmol kg-1, i.v.) to conscious rats resulted in a prolonged pressor effect preceded by a transient depressor response. Corresponding to changes in blood pressure, a small transient tachycardia was followed by a sustained bradycardia. ET-l enhanced albumin leakage by 87 and 120% in the left ventricle and right atrium, respectively, as measured by the extravasation of Evans blue dye.5. The selective ETA receptor antagonist, FR 139317 (2.5 mg kg-1) significantly blunted the pressor action of ET-1 and the accompanying bradycardia without affecting the depressor response. Furthermore,FR 139317 almost completely abolished the permeability effect of ET-l in both vascular beds studied.6. Pretreatment of the animals with BM 13505 (1 mg kg-1), OKY-046 (10mg kg-1), WEB 2086(1 mg kg-1) or BN 52021 (10mg kg-1) significantly reduced ET-1 (1 nmol kg-1)-induced albumin extravasation both in the left ventricle and right atrium. The PAF receptor antagonists, WEB 2086 and BN 52021 were equally potent inhibitors in the left ventricle, whereas BN 52021 appeared to be a more potent inhibitor than WEB 2086 in the right atrium. Pretreatment with indomethacin (3 mg kg-1 plus 2 mg kg-1 h-1) did not modify the permeability response to ET-1. None of these compounds affected significantly ET-l-induced changes in mean arterial blood pressure and heart rate.7. These results indicate that intravenous administration of ET-1 provokes ST segment elevation and myocardial oedema and suggest that these events are mediated, in part, through release of secondary mediators, such as PAF and TxA2 via the activation of ETA receptors.


Are women with breast cancer more likely to develop colorectal cancer? Critical review and meta-analysis.
Breast cancer has been reported to confer greater risk for colorectal cancer. In order to estimate the magnitude of this risk, we critically reviewed all studies evaluating the association of breast and colorectal cancer and performed a meta-analysis. The combined results from seven cohort studies demonstrated a weak association between breast cancer and the subsequent risk of colorectal cancer [pooled relative risk (RR) = 1.15; 95% confidence interval (CI) = 0.99-1.31]. Pooled results from five cohort studies showed that the risk of breast cancer after colorectal cancer was similar (pooled RR = 1.10; 95% CI = 1.03-1.17). The combined results from five cross-sectional/case-control studies revealed a positive association between breast cancer and colorectal adenomas (pooled RR = 1.74; 95% CI = 1.27-2.21). Population-based cohort studies, the strongest research design, show essentially no increase in risk of colorectal cancer in women with previous breast cancer. Based upon this evaluation, there appears little justification for special colorectal surveillance of women with previous breast cancer.


Dosimetric verification of the 95% isodose surface for a conformal irradiation technique.
In the treatment planning of conformal radiotherapy, field shapes are often designed in such a way that a high-value isodose surface fully encompasses the target volume. Therefore, knowledge about the accuracy with which the treatment planning system calculates the position of that isodose surface is essential to prevent field shapes which are either too large or too small. To determine this accuracy for a conformal multi-field technique, the dose in the high-dose region must be measured with a high spatial resolution. A method is presented to reconstruct and evaluate the experimental high-dose region from a set of water phantom scans. This method, which assesses combined dose profiles for multi-field irradiation techniques, can be used for the commissioning and/or quality assurance of a 3-D treatment planning system. For a specific conformal technique, the measured and calculated 95% isodose positions along lines in several directions have been compared. It is shown that different dose values of single beam profiles determine the resulting 95% isodose position, which is important to recognize for quality assurance of treatment planning calculations. It is further found that the uncertainty in the calculated 95% isodose surface can be described by a standard deviation in dose value, which relates to a positional uncertainty through the local dose gradient. Thus the confidence region of the calculated 95% isodose can be indicated in the treatment plan by plotting isodoses at the 95% level plus and minus its standard deviation. Such a procedure is recommended instead of plotting the 95% isodose with a constant width. In addition, restrictions for the cumulative dose-volume histogram of acceptable treatment plans can be formulated, based on the sensitivity of the actual target coverage on the uncertainty with which the prescribed isodose surface is calculated.


Military suicide coverage: AP news wire & suicide guidelines.
The authors evaluated how well Associated Press News Wire stories adhered to the Recommendations for Reporting on Suicide™ during 2012, a peak year of military suicide. They included individual suicide stories (N = 167) from randomly selected days. They also evaluated differences in the military versus civilian coverage. Military and civilian stories typically had about five negative practices, and less than one positive practice, with military stories significantly more likely to miss valuable opportunities to promote help-seeking. The findings, combined with previous evidence, suggest the need for the development of specific military suicide reporting guidelines.


Insomnia as a sequela of sarin toxicity several years after exposure in Tokyo subway trains.
More than 5,000 passengers on Tokyo subway trains were injured with toxic chemicals including the nerve gas "sarin" on March 20, 1995. The purpose of this study was to identify the effect of sarin exposure on insomnia in a cross-sectional study. A self-administered questionnaire concerning sleep-related items was distributed to victims of sarin exposure in October and November, 2003. Questionnaires were completed by 161 of the 163 participants (98.8%), who were selected from 1,500 subjects. Among them, the authors selected 75 women 30 to 69 years of age. Control participants were collected from inhabitants living in Maebachi City, Gunma Prefecture, Japan. For the younger exposed group (under 50 yr. of age), percentages of poor sleep, difficulty falling asleep, intermittent awakening, early morning awakening, a feeling of light overnight sleep, and insomnia were significantly higher than those for the control group. In contrast, the older exposed group (ages 50 to 69 years) had significantly higher prevalence of poor sleep, a feeling of light overnight sleep, and early morning awakening for the exposed group when compared with the control group. The high prevalence of insomnia and insomnia-related factors for victims especially under 50 years of age suggests a need for research on sleep quality after sarin exposure. Although posttraumatic stress disorder is assumed to be a psychological effect of exposure to a toxic substance, a cause-and-effect relationship has not been established.


[Necrotizing fasciitis].
The management of a right foot necrotizing fasciitis and severe sepsis in an old diabetic patient is presented. The early and aggressive surgery, adequate antibiotherapy and correction of organic disfunction (cardio-circulatory, renal, respiratory) eliminated the vital risk and resulted in a satisfactory morpho-functional recovery of the foot.


Closure of the greater saphenous vein with endoluminal radiofrequency thermal heating of the vein wall in combination with ambulatory phlebectomy: 50 patients with more than 6-month follow-up.
Incompetence of the saphenofemoral junction (SFJ) with reflux into the greater saphenous vein (GSV) is one cause of chronic venous hypertension that may lead to the development of varicose and telangiectatic leg veins. To evaluate with long-term follow-up a novel method for closing the incompetent GSV at its junction with the femoral vein through an endoluminal approach. Fifty patient legs with reflux at the SFJ into the GSV were treated with radiofrequency (RF) heating of the vein wall through an endoluminal catheter. Patients were evaluated at 6, 9, 12, and 18-24 months to determine treatment efficacy as well as adverse sequelae. Nine patients could not return for evaluation after 3 months. The remaining 41 patient legs were evaluated. Sixty-eight percent of treated legs achieved complete closure of the SFJ and GSV. Twenty-two percent of legs had an open GSV without reflux. Three patients (four legs) (10%) had an open and refluxing GSV. Three of 41 legs had recurrent varicose veins. Only 1 of 41 legs had recurrent symptoms. Complete treatment took an average of 20 minutes. Adverse sequelae were minimal, with 28 legs showing purpura less than 2 weeks, 5 legs with erythema less than 1 week, and 8 legs with an indurated fibrous cord lasting for up to 6 months posttreatment. Endoluminal RF thermal heating of an incompetent GSV has been shown to be easily accomplished and efficacious throughout the 24-month-follow-up period.


Grouper (Epinephelus coioides) TCR signaling pathway was involved in response against Cryptocaryon irritans infection.
T cell activation is a complicated process accompanying with the activation of T cell receptor (TCR) signaling pathway, which is not well described in teleost fish. The initiation of this pathway depends on the interaction of membrane TCR co-receptors (e.g. CD4/8, CD3 and CD45) and a series of cytoplasmic protein tyrosine kinases (e.g. Lck, Fyn and ZAP70). Cyptocaryon irritans is a ciliate pathogen of marine fish white spot disease causing huge economic lost in marine aquaculture. This parasite can infect fish gill and skin and is considered to be a good pathogen model for fish gill and skin mucosal immunity. Our previous studies showed the locally mucosal antibody response was important for fish defense against this parasite. While how TCR signaling pathway involved in T cell activation to help B cell activation in C. irritans infected fish is still not known. In the present study, we cloned a grouper TCR co-receptor gene EcCD3ε (537 bp) and its three kinase genes, including EcLck (1512 bp), EcFyn (1605 bp) and EcZAP70 (1893 bp). Homology analysis showed that they all shared the highest identity with corresponding genes from Takifugu rubripes (EcCD3ε 41%, EcLck 88%, EcFyn 98% and EcZAP70 93%), and their conserved motifs involved in the signaling transduction were analyzed. The tissue distribution analysis showed these four genes were high expressed in thymus, and it is interesting to find their comparative high expression in skin, gill and midgut mucosal immune tissues. In C. irritans infected grouper, the expression of three TCR co-receptors (EcCD4-1, EcCD3ε and EcCD45) and three kinases (EcLck, EcFyn and EcZAP70) was tested in skin, gill, head kidney and spleen at 0, 12 h, 24 h, 2 d, 3 d, 5 d and 7 d. All six genes were significantly up-regulated in skin at most tested time points, which indicate the possibility of skin local T cell activation to support the local antibody response. Compared to three TCR co-receptors, significantly up-regulation of three kinases were seen in the spleen, and the spleen fold changes of these three kinases were much higher than head kidney, which indicates spleen maybe the major systematic immune organs for T cell activation in C. irritans infected fish.


Dendritic reticulum cell-related immunostaining for laminin in follicular and diffuse B-cell lymphomas.
We performed a comparative immunohistocytochemical study of the distribution patterns of laminin and follicular dendritic reticulum cells (DRCs) within their follicular microenvironment in both nodular or diffuse B-cell non Hodgkin's lymphomas (NHLs). Twenty nine cases of immunophenotypically diagnosed B-cell NHLs (19 of follicular center cell origin-FCCL- and 10 of the diffuse well differentiated lymphocytic type-WDLL-) and five reactive lymph nodes with follicular hyperplasia were analyzed by immunoperoxidase and immunofluorescence techniques. Serial frozen sections and cytospin preparations were tested either with single antibodies anti laminin and DRC-1, or paired reagents in double labeling immunofluorescence. Our results indicated consistently that within both the reactive germinal centers and the neoplastic nodules of FCCL laminin immunostaining visualized a punctate-granular pattern apart from the linear vascular basement membrane positivity. Double immunofluorescence assay demonstrated that there was a close parallelism between this laminin staining pattern and DRC-1 distribution showing a well developed DRCs meshwork; in the diffuse tumour areas of both FCCL and WDLL, laminin immunoreactivity was found only in those cases in which nests of DRCs were observed. Double immunofluorescence studies performed on cytospin preparations demonstrated that the groups of cells containing DRC-1 positive cells, contained a positivity for laminin, although within the cell the staining for DRC-1 was intense and diffuse, while that for laminin was granular and more sparse. Our results suggested that these laminin and DRC-1 positive reactive sites may be present on the same cells. Since the reduction in number or loss of both DRCs and their related immunostaining for laminin within the microenvironment was consistently associated with a loss of nodularity by lymphoma cells, whereas nodularity in reactive and neoplastic conditions was associated with a rich DRCs meshwork and the related laminin immunostaining, a trapping function of DRCs exercised in the presence of laminin should be considered.


An imaging technique for detection and absolute calibration of scintillation light.
Triggered by the need of a detection system to be used in experiments of nuclear fusion in laser-generated plasmas, we developed an imaging technique for the measurement and calibration of the scintillation light yield of scintillating materials. As in such experiments, all the reaction products are generated in an ultrashort time frame, the event-by-event data acquisition scheme is not feasible. As an alternative to the emulsion technique (or the equivalent CR39 sheets) we propose a scintillating screen readout by means of a high performance charge coupled device camera. Even though it is not strictly required in the particular application, this technique allows the absolute calibration of the scintillation light yield.


Oxidative Stress, Cytotoxicity, and Genotoxicity Induced by Methyl Parathion in Human Gingival Fibroblasts: Protective Role of Epigallocatechin-3-Gallate.
Organophosphorous (OP) compounds are pesticides frequently released into the environment because of extensive use in agriculture. Among these, methyl parathion (mPT) recently received attention as a consequence of illegal use. The predominant route of human exposure to mPT is via inhalation, but inadvertent consumption of contaminated foods and water may also occur. The goal of this study was to investigate the in vitro effects of mPT on cells in the oral cavity and evaluate the potential protective role of epigallocathechin-3-gallate (EGCG) on these effects. Human gingival fibroblasts (HGF) were exposed to 10, 50, or 100 μ g/ml mPT for 24 h and assessed for oxidative stress, as evidenced by reactive generation of oxygen species (ROS), induction of apoptotic cell death, DNA damage (comet assay and cytochinesis-block micronucleus test), and nitric oxide (NO) production. The results showed that mPT produced significant oxidative stress, cytotoxicity, and genotoxicity and increased NO levels through stimulation of inducible NO synthase expression. Finally, data demonstrated that EGCG (10, 25, or 50 μ M) was able to inhibit the pesticide-induced effects on all parameters studied. Data indicate that cytotoxic and genotoxic effects may be associated with oxidative stress induced by mPT observed in HGF cultures and that EGCG plays a protective role via antioxidant activities.


[Murine pulmonary fibrosis model induced by repeated low-dose intravenous injection and intratracheal instillation of bleomycin].
To determine the characteristics and differences in bleomycin-induced lung fibrosis model by repeated low-dose intravenous injection and single dose intratracheal instillation of bleomycin. Forty male ICR (Institute for Cancer Research) mice were randomly divided into a model group I, a model group II, and 2 control groups (10 mice in each group). In model group I, bleomycin was injected intravenously at 10 mg/(kg.d) for 14 consecutive days; and in model group II, bleomycin was instilled intratracheally at 5 mg/kg. The 2 control groups were given isotonic saline solution. At the 28th day, the mice were sacrificed and the bronchoalveolar lavage fluid (BALF) was collected. The total cells and proteins in the BALF, pulmonary coefficient, and hydroxyproline (HYP) content were determined. The pathological changes were observed by the eosin staining and Masson's trichrome staining. 1) Both intravenous injection and intratracheal instillation of bleomycin resulted in severe and extensive inflammation and fibrosis in the lungs. The total cells and proteins in the BALF, HYP content, pulmonary coefficient and the pathological score of pulmonary fibrosis were all significantly increased in the 2 model groups (P<0.01). 2) Fibrosis was mainly under the pleura or around the vessel in model group I, and it was located near the bronehia and bronchioles in model group II. 3) The death rate was higher in the model group II than that in the model group I. 4) Proteins in the BALF were significantly higher in model group II than that in model group I (P<0.05). There was no difference in the total cells in the BALF, the pulmonary coefficient, the HYP content, and the pathological score of pulmonary fibrosis between the 2 groups (P>0.05). The pulmonary fibrosis model can be successfully established by intravenous injection or intratracheal instillation of bleomycin, but the sites of pulmonary fibrosis are different. The histological changes caused by the repeated low-dose intravenous injection of bleomycin is more similar to idiopathic pulmonary fibrosis than that by the single dose intratracheal instillation.


Disinhibition in the human motor cortex is enhanced by synchronous upper limb movements.
The phasic modulation of wrist flexor corticomotor disinhibition has previously been demonstrated during the flexion phase of rhythmical passive flexion-extension of the human wrist. Here we ask if rhythmical bimanual flexion-extension movements of the wrists of neurologically intact subjects, modulate inhibitory activity in the motor cortex. In the first experiment intracortical inhibition was assessed when one wrist was passively flexed and extended on its own, with the addition of the opposite limb voluntarily moving synchronously in a mirror symmetric pattern, and also in a near-symmetric asynchronous pattern. Two subsequent experiments investigated firstly the modulation of spinal reflex pathway activity during the same three movement conditions, and secondly the effect of contralateral wrist movement alone on the excitability of corticomotoneuronal pathways to a static test limb. When the wrist flexors of both upper limbs were shortening simultaneously (i.e. synchronously), intracortical inhibition associated with flexor representations was suppressed to a greater extent than when the two muscles were shortening asynchronously. The results of the three experiments indicate that modulation of inhibitory activity was taking place at the cortical level. These findings may have further application in the study of rehabilitation procedures where the effects of simultaneous activation of affected and unaffected upper limbs in hemiparetic patients are to be investigated.


Adipose conversion of mouse bone marrow fibroblasts in vitro: their alkaline phosphatase activity.
By an in vitro colony assay and cytochemical staining, we investigated the capacity of mouse bone marrow fibroblasts to differentiate into adipocytes and to express alkaline phosphatase (ALP) activity. Glucocorticoids enhanced colony formation of the fibroblasts, and stimulated their adipose conversion (55-65% of the colonies became adipocyte-positive), but they did not affect ALP activity. The fibroblasts became heterogeneous in size and morphology after growing in vitro then differentiated into adipocytes. All the cell types had ALP activity, and more than 95% of the colonies contained ALP-positive cells. ALP staining was strongest in cells in the early stage of adipose conversion, gradually decreasing with maturation. Our results indicate that the majority of the mouse bone marrow fibroblasts that formed colonies under our culture conditions are preadipocytes. We conclude that these fibroblasts originate from adventitial reticular cells present in bone marrow stroma because reticular cells have been reported to possess high ALP activity and have been suggested to differentiate into adipocytes.


Suggestions for improving the study of health program implementation.
More will be learned about health programs and the implementation of health policy in this country if we pay more attention to issues of program implementation. Of particular use would be more studies which explicitly link program implementation with program outcomes and which recognize the need to combine quantitative and qualitative analysis of program implementation; the use of triangulated methods in focusing on the relationship between program implementation and program outcomes; the incorporation and study of planned variation in the methods of implementing programs; recognition that the process is essentially one of organizational change and innovation, and the incorporation of existing theory and evidence relevant to these issues; and recognition that the ongoing nature of the implementation process requires longitudinal study designs for implementation as well as for outcome assessment. Cronbach [9] has remarked that evaluation research "lights a candle in the darkness, but it never brings dazzling clarity." It may be that more attention to program implementation and better research on the process, such as that suggested in this note, will provide a little more light and will bring if not dazzling , at least modest, improvements in clarity.


Fulminant liver failure after radical prostatectomy in the hyperlordotic position.
In the hyperlordotic position, the patient's hips are above the feet and head. This position has previously been associated with partial vena caval occlusion, increased venous pressures, and spinal cord infarction. We describe a case of fulminant hepatic failure after radical retropubic prostatectomy performed in the hyperlordotic position.


Reciprocal EMG control of elbow extension by FES.
Elbow extension is critical in performing activities of daily living. Individuals with a C5-C6 spinal cord injury have paralyzed elbow extensors, yet retain weak to strong voluntary control of elbow flexion. Previous studies have shown that functional electrical stimulation (FES) of the triceps provides sufficient elbow extension strength and control to greatly improve function. With triceps stimulation applied at a constant level, elbow angle is controlled naturally by voluntary flexion opposing the stimulated extension-referred to as voluntary antagonist control. We have investigated an alternative reciprocal control scheme employing biceps electromyogram (EMG) to modulate triceps stimulation. With reciprocal control, increasing biceps EMG proportionally reduces triceps stimulation. A personal computer (PC)-based lab system was designed to test the feasibility of reciprocal control. Reciprocal control increased the range of elbow moments, was stable during maintained elbow angle or isometric moment, and used less stimulation. Reciprocal control of triceps stimulation using biceps EMG is an effective method for restoring elbow extension to C5-C6 spinal cord injury patients, and could be extended to other situations where a voluntarily controlled muscle can be opposed by stimulating an antagonist.


[The complex travel of cinchona barks between Peru and Québec Hôtel-Dieu, in the middle of the eighteenth century].
Duplessis sisters, who were religious nuns in Hôtel-Dieu (Quebec hospital), were exchanging letters with a French apothecary from Dieppe in Normandy, named Jacques-Tranquilain Féret. They asked him to send them in Quebec the drugs and medicines their apothecary needed. Amongst these drugs were cinchona barks that came from Callao in Peru by boat, passed Cape Horn and then sailed to Cadiz, the great Spanish port. Then they embarked to Rouen, which was the French port for goods coming from overseas. The goods from Peru had then to be transported on little fishing boats to Dieppe, where Féret received the barks. The apothecary sent these drugs to Quebec by boats sailing either from Rouen or from La Rochelle. So these Peruvian drugs had to cross two times the Ocean before accessing to North America.


Flexible Wearable Capacitive Sensors Based on Ionic Gel with Full-Pressure Ranges.
Flexible positive pressure sensors have been studied extensively and have been used in a lot of scenarios. However, negative pressure detection is also in demand in some scenarios, such as fluid mechanics analysis, air pressure sensing, and so on. Flexible wearable sensors that can detect both positive and negative pressures will greatly broaden the application field. In this paper, we report a flexible highly sensitive ionic gel (IG) pressure sensor, which is simple and of low cost to prepare and can reliably detect a large pressure range from -98 to 100 kPa under an atmospheric pressure of about 982 hPa. The IG dielectric layer is composed of polyvinyl alcohol and phosphoric acid with a random microstructure of sandpaper inversion. The sensor exhibits flexibility, cycling stability, and high sensitivity under both negative and positive pressures (S = 84.45 nF/kPa for the negative pressure section, S = 25.61 nF/kPa for the positive pressure section). These sensors could be worn on the body not only to test breathing and pulse but also to measure air pressure for estimating the altitude, showing that the flexible full-pressure sensors have a wider application range in wearable electronics.


The nurse director role is more challenging than ever.
I was a nurse director for more than 20 years, initially at Royal Brompton Hospital, then at Royal Brompton and Harefield NHS Foundation Trust. It was a complex role and an overwhelmingly positive experience. Over the years, however, much has changed, and today the average tenure is about three years.


An update on Experimental and Clinical Psychopharmacology: Something old, something new, something borrowed, something green?
In this editorial, the author provides an update on Experimental and Clinical Psychopharmacology in several areas. First, the journal will continue to accept original research reports and full reviews as it has in past years. The author hopes to still receive outstanding manuscripts in the journal's primary areas of strength, such as clinical research on alcohol use and cigarette smoking. The journal will also continue to publish an annual special issue on a current topic in the field. Second, the journal now accepts brief communications, brief reviews, and case reports. The authors sees these new formats as opportunities to publish cutting edge, novel findings that may not be suitable as original research reports or full reviews-such work would previously not have fit with the journal. Third, the author has borrowed an idea from colleagues who serve as editors for other journals in the field: the addition of an editorial fellowship at Experimental and Clinical Psychopharmacology. Finally, the "something green" part of the title refers to the new, bright green cover color of the print version of the journal. (PsycINFO Database Record


Vulval hidradenoma papilliferum: a clinical and dermoscopic study.
Hidradenoma papilliferum (HP) is a rare, slow-growing, benign adnexal tumour with apocrine differentiation. It usually affects the anogenital region in adult women and is clinically polymorphous, mimicking other benign or malignant neoformations. The dermoscopic features of vulval HP have not been reported yet. We report the clinical and dermoscopic features in a case series of histopathologically proven vulval HPs. Dermoscopy may be a useful tool for the diagnosis of vulval HP. To our knowledge, our paper represents the first report of dermoscopic findings in a series of vulval HPs in a multiracial population.


The role of mononuclear phagocytes in Ebola virus infection.
The filovirus, Zaire Ebolavirus (EBOV), infects tissue macrophages (Mϕs) and dendritic cells (DCs) early during infection. Viral infection of both cells types is highly productive, leading to increased viral load. However, virus infection of these two cell types results in different consequences for cellular function. Infection of Mϕs stimulates the production of proinflammatory and immunomodulatory cytokines and chemokines, leading to the production of a cytokine storm, while simultaneously increasing tissue factor production and thus facilitating disseminated intravascular coagulation. In contrast, EBOV infection of DCs blocks DC maturation and antigen presentation rendering these cells unable to communicate with adaptive immune response elements. Details of the known interactions of these cells with EBOV are reviewed here. We also identify a number of unanswered questions that remain about interactions of filoviruses with these cells.


[Acute gastro-enteritis in children in France: estimates of disease burden through national hospital discharge data].
To estimate the burden of hospitalized infectious gastroenteritis of children younger than 5 years of age and associated costs. We analyzed 1997 hospital discharges with a primary diagnosis of gastroenteritis or a secondary diagnosis of gastroenteritis with gastroenteritis symptoms or complications as primary diagnosis and compared the deaths with those of the national mortality data. Gastroenteritis was associated with 51,125 hospitalizations which accounted for 11.4% of hospitalization discharges for this age group and an annual rate of 1,385 per 100,000 children <5-year-old. Most gastroenteritis (56%) were registered as "probably infectious", 36% as "viral" (43% of which were coded "rotavirus") and 8% as "bacterial" (of which 60% were coded "Salmonella"). The seasonal peak was winter for rotaviral, viral and "probably infectious" gastroenteritis, summer for those related to salmonellosis. Incidence increased inversely with age: 3606/100,000 infants <1-year-old, 257/100,000 4-year-old children. Complications (especially dehydration) were observed in 21% of viral gastroenteritis and 17% of bacterial gastroenteritis. At least, 14 deaths were found in both hospital discharge and mortality data. The mean duration of stay (3.2 days) was significantly higher in infants <1-year-old, viral etiology, association with complications or bronchiolitis. The costs of hospitalization could be estimated to 62 million Euros. Our results are similar to those obtained in other developed countries. Despite variations in encoding the discharge reports, data has proven to be effective to describe national trends for this health event. Our study indicates that the public health burden and economic impact of prevention and control measures can be monitored through hospital discharge surveillance.


Bipolar disorder preceded by substance abuse: a different phenotype with not so poor outcome?
The co-occurrence of bipolar disorder and substance use is frequent. The question whether substance use precedes, induces or follows bipolar disorder is still unresolved. Substance use has been typically represented as a negative prognostic factor for the clinical course of bipolar illness and it has been associated with decreased compliance and treatment resistance. To extend these previous findings we examined the clinical course and outcome of patients with bipolar disorder I or II whose bipolar onset was preceded by substance/alcohol use. The impact of substance use as a predisposing or precipitating factor of a bipolar episode was evaluated by comparing a group of bipolar subjects whose mood symptomatology onset was preceded by substance/alcohol use (N=145) (SUBP) with a similar sized representative sample of bipolar patients (either substance users/abusers or not using substances) whose first mood episode was unrelated to substance use (N=144) (NSUBP). RESULTS; The analysis of clinical and socio-demographic variables revealed that patients in the SUBP group showed less severe psychopathology, as indicated by fewer total hypomanic and depressive episodes during the course of their illness. Depressive onset was less frequent amongst the SUBP group. SUBP patients, compared to NSUBP, were poorly compliant to treatment. Bipolar disorder preceded by substance misuse may represent a clinically milder subtype of bipolar illness. This subtype would be less "primary" and might be more early targeted by primary prevention with programmes focused on substance misuse.


Intracellular ATP levels determine cell death fate by apoptosis or necrosis.
Although apoptosis and necrosis are morphologically distinct manifestations of cell death, apoptosis and some necroses share common features in the death signaling pathway involving functional steps of death-driving interleukin 1beta-converting enzyme family proteases and anti-cell death protein Bcl-2. One evident physiological difference in cells undergoing apoptosis versus necrosis is in intracellular levels of ATP. In this study, we specifically addressed the question of whether apoptosis depends on intracellular ATP levels, since longer incubation under ATP-depleting conditions results in necrotic cell death. Incubation of cells in glucose-free medium with an inhibitor of mitochondrial F0F1-ATPases reduces intracellular ATP levels and completely blocks Fas/Apo-1-stimulated apoptosis. ATP supplied through glycolysis or oxidative phosphorylation restores the apoptotic cell death pathway. ATP depletion also leads to a block in Fas-induced activation of CPP32/Yama(-like) proteases, and when ATP is depleted after the activation of the proteases, subsequent apoptosis is significantly blocked. Thus, ATP-dependent steps exist both upstream and downstream of CPP32/Yama(-like) protease activation in apoptotic signal transduction. Treatment with the calcium ionophore induces apoptosis under ATP-supplying conditions but induces necrotic cell death under ATP-depleting conditions, indicating that ATP levels are a determinant of manifestation of cell death.


Recall and recognition hypermnesia for Socratic stimuli.
In two experiments, we investigate hypermnesia, net memory improvements with repeated testing of the same material after a single study trial. In the first experiment, we found hypermnesia across three trials for the recall of word solutions to Socratic stimuli (dictionary-like definitions of concepts) replicating Erdelyi, Buschke, and Finkelstein and, for the first time using these materials, for their recognition. In the second experiment, we had two "yes/no" recognition groups, a Socratic stimuli group presented with concrete and abstract verbal materials and a word-only control group. Using signal detection measures, we found hypermnesia for concrete Socratic stimuli-and stable performance for abstract stimuli across three recognition tests. The control group showed memory decrements across tests. We interpret these findings with the alternative retrieval pathways (ARP) hypothesis, contrasting it with alternative theories of hypermnesia, such as depth of processing, generation and retrieve-recognise. We conclude that recognition hypermnesia for concrete Socratic stimuli is a reliable phenomenon, which we found in two experiments involving both forced-choice and yes/no recognition procedures.


Maternal depression and parental distress among families in the Early Head Start Research and Evaluation Project: Risk factors within the family setting.
This study examined the influence of risk factors within the family environment (inadequate resources, insufficient caregiving support from child's father, higher family conflict) on maternal well being (depression and parental distress) using a longitudinal panel research design. Participants consisted of 2,040 low-income mothers of young children enrolled in the Early Head Start (EHS) Research and Evaluation Project. Findings indicated that greater family resources at 14 months had a protective effect on maternal well-being at 36 months, controlling for demographic variables and maternal well-being at 14 months. Furthermore, the effect of family resources on maternal well-being was moderated by father caregiving support, such that family resources were protective, particularly when father caregiving support was lower. Family conflict at 14 months was associated with lower maternal well-being at 36 months, controlling for demographic variables and maternal well-being at 14 months. Our findings suggest that low-income mothers need the support and resources of programs such as EHS to overcome the hurdles and challenges of parenting in high-risk settings. Implications for clinical practice and EHS are discussed.


The Intranodal Presence of Coexisting Granulomatous Inflammation and Carcinoma During Transbronchial Needle Aspiration of Intrathoracic Lymphadenopathy.
The presence of intrathoracic lymphadenopathy in patients with suspected malignancy remains concerning, often prompting further evaluation with tissue sampling. The presence of nodal granulomatous inflammation in patients with underlying malignancy is well reported. However, review of 3 recent large trials of endobronchial ultrasound-guided transbronchial needle aspiration in patients with granulomatous inflammation and malignancy did not identify the presence of coexisting, intranodal malignancy, and granulomatous inflammation, rather these diagnoses remained nodally exclusive. We present a case of coexisting granulomatous inflammation and metastatic carcinoma within the same lymph node aspirates, reviewing the potential diagnostic pitfalls and implications of this rare occurrence.


[Phosphate binders : What are the recent evidences ?]
Hyperphosphatemia is a late complication of chronic kidney disease (CKD). It is a major non-traditional cardiovascular risk factor in CKD. The management of hyperphosphatemia is based on restriction of phosphate intake, phosphate binders and dialysis. Phosphate binders are divided into calcium-based and non-calcium-based binders. Recent evidence suggest restricting the dose of calcium-based phosphate binders because of the associated increased mortality in compared to non-calcic ones. New treatments are under development such as inhibitors of kidney or intestinal phosphate absorption. These therapies could improve patients' compliance.


[Clinical experience with sumatriptan ++ in the treatment of migraine symptoms].
Although sumatriptan has proven effective in treating migraine symptoms, there have been no studies comparing this drug with non-steroid anti-inflammatory drugs (NSAIDs) and ergotamine compounds under ideal dosage and delivery conditions. Our study aimed to establish the objective and subjective effects of treatment of severe migraine attacks with subcutaneous sumatriptan in a series of patients who were resistant to or intolerant of NSAIDs or ergotamine derivatives. Twenty-three patients were treated for a total of 140 attacks of migraine (mean +/- SD = 6 +/- 4 attacks). In 26% of the patients and in 23% of the attacks the response to sumatriptan was inadequate, while the response was clearly positive in 74% of the patients and in 77% of the attacks. More than half the patients responded within the first 30 min after injection of the drug. In 52% side effects were reported, although none were severe. Side effects continued with the same intensity or decreased in 91% of the patients in the course of the study. No patient considered treatment with sumatriptan worse than treatment with ergotamines or NSAIDs, while 86% and 91% labelled sumatriptan as "better" or "much better" than NSAIDs and ergotamines, respectively. Eighteen (78%) stated that they would continue using sumatriptan for future migraine attacks. Finally, most considered price to be a factor to be taken into consideration in deciding whether to inject the drug indiscriminately.(ABSTRACT TRUNCATED AT 250 WORDS)


Intracemetery biological variation at the fort ancient sunwatch village.
We investigate intracemetery biological variation at the Fort Ancient SunWatch village (800-500 BP.) to test the hypothesis that the SunWatch population consisted of spatially structured biologically differentiated kin groups consisting of distinct local Late Woodland and non-local Mississippian biological populations. The SunWatch sample contains 166 individuals: 63 adults and 103 subadults. We analyze intracemetery biological variation using two feature sets: the buccolingual diameters of the polar teeth in the permanent (n = 37) and deciduous (n = 26) dentitions. We apply matrix correlation models to biodistance and grave distance matrices in both data sets, evaluate burial outliers and individuals exhibiting a rare dental feature with interindividual biological distances, and evaluate potential cultural subgroups with Mahalanobis' distance. Matrix correlation analyses for both feature sets as well as interindividual and subgroup distances indicate the SunWatch village cemetery was kin structured, contained a single primary biological population, and also contained a small number of non-local individuals most of whom were from closely related populations. We thus reject the hypothesis that the SunWatch population consisted of biologically distinct Late Woodland and Mississippian biological populations. Although SunWatch village exhibited cultural attributes characteristic of both local Late Woodland and non-local Mississippian groups, biologically the village was composed of one primary population. The lack of evidence for marked biological differentiation in the SunWatch village area at about 800-500 BP. suggests diffusion and acculturation may account for Mississippian cultural characteristics in southwestern Ohio Fort Ancient villages. If gene flow or migrations from biologically distinct Mississippian populations into southwestern Ohio occurred, either or both likely occurred well before 800 BP. This would have allowed the process of admixture time to produce a relatively homogeneous, nondifferentiated population. The latter alternative is consistent with the appearance in southwestern Ohio of non-local individuals at one site linked to surrounding Mississippian regions at about 950 BP. Am J Phys Anthropol 160:719-728, 2016. © 2016 Wiley Periodicals, Inc.


Understanding the role of root-related traits in salinity tolerance of quinoa accessions with contrasting epidermal bladder cell patterning.
To compensate for the lack of capacity for external salt storage in the epidermal bladder cells, quinoa plants employ tissue-tolerance traits, to confer salinity stress tolerance. Our previous studies indicated that sequestration of toxic Na+ and Cl- ions into epidermal bladder cells (EBCs) is an efficient mechanism conferring salinity tolerance in quinoa. However, some halophytes do not develop EBCs but still possess superior salinity tolerance. To elucidate the possible compensation mechanism(s) underlying superior salinity tolerance in the absence of the external salt storage capacity, we have selected four quinoa accessions with contrasting patterns of EBC development. Whole-plant physiological and electrophysiological characteristics were assessed after 2 days and 3 weeks of 400 mM NaCl stress. Both accessions with low EBC volume utilised Na+ exclusion at the root level and could maintain low Na+ concentration in leaves to compensate for the inability to sequester Na+ load in EBC. These conclusions were further confirmed by electrophysiological experiments showing higher Na+ efflux from roots of these varieties (measured by a non-invasive microelectrode MIFE technique) as compared to accessions with high EBC volume. Furthermore, accessions with low EBC volume had significantly higher K+ concentration in their leaves upon long-term salinity exposures compared to plants with high EBC sequestration ability, suggesting that the ability to maintain high K+ content in the leaf mesophyll was as another important compensation mechanism.


Optimizing therapy of seizures in stroke patients.
Stroke is the leading cause of symptomatic epilepsy in adults, accounting for up to one-third of newly diagnosed seizures among the elderly. About 3% to 5% of stroke patients will suffer a remote seizure, 54% to 66% of whom will develop epilepsy. Thus far, the optimal timing and type of antiepileptic treatment for patients with post-stroke seizure and epilepsy have not been specifically assessed. Although several studies suggest that seizures alter the functional recovery after a stroke, it remains difficult to determine whether or not the occurrence of a second seizure in an untreated stroke patient might hamper the overall outcome. The decision to initiate antiepileptic drug (AED) treatment after a first or a second post-stroke seizure should therefore be individualized, primarily based on the functional impact of the first seizure episode and the patient's preference. Several converging findings suggest that the majority of first-generation AEDs, particularly phenytoin, are not the most appropriate choice in stroke patients because of their potential harmful impact on functional recovery and bone health, their suboptimal pharmacokinetic profile and interaction with anticoagulants or salicylates, their greater likelihood to be poorly tolerated, and the lack of level A evidence regarding their specific use in elderly patients. Among the new-generation AEDs that do not interact with anticoagulants, antiplatelet agents, or bone health, lamotrigine and gabapentine are the only two drugs that proved to be more effective than immediate-release carbamazepine in elderly patients, providing level A evidence for their use in this indication. In addition, gabapentin remains the only drug that has been specifically evaluated in stroke patients, demonstrating a high rate of long-term seizure freedom. At present, low-dose lamotrigine or gabapentin appears to represent the optimal first-line therapy for post-stroke seizure and epilepsy in elderly patients or in younger patients requiring anticoagulants. However, low-dose extended-release carbamazepine might be a reasonable and less expensive option in patients with appropriate bone health who do not requiring anticoagulation.


Serum withdrawal leads to reduced aryl hydrocarbon receptor expression and loss of cytochrome P4501A inducibility in PLHC-1 cells.
Changes in the expression of the aryl hydrocarbon receptor (AHR) have been documented in several systems and in response to a variety of treatments. The significance of these findings is unclear, because the effects of such changes on subsequent responses to AHR ligands seldom have been measured. We tested the ability of changes in serum used in cell culture medium to alter expression of the AHR and induction of cytochrome P4501A (CYP1A) in PLHC-1 teleost hepatoma cells. Culture of early-passage cells in serum-free medium for 2 days led to a loss of CYP1A inducibility by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In contrast, culture in 10% delipidated calf serum increased the TCDD-induced levels of both CYP1A protein and enzymatic activity relative to levels in cells cultured in 10% complete calf serum. These effects were consistent between 8 and 24hr post-treatment, indicating that the kinetics of induction were unaffected. In cells cultured in serum-free medium for 1 and 2 days there was a progressive loss of CYP1A inducibility. This loss of response paralleled a time-dependent decline in AHR protein, as measured by specific binding of [3H]TCDD. Using an operational model for AHR action in PLHC-1 cells, the measured reduction in AHR could be shown to predict the loss of CYP1A induction. Expression of AHR protein was unaffected by culture in 10% delipidated serum. The effects of serum-free medium and delipidated serum were found only in early-passage cells; inducibility of CYP1A and expression of AHR protein in late-passage cells were unaffected by serum withdrawal. Comparison of early- and late-passage cells revealed a 2-fold greater rate of proliferation in the latter, suggesting that a growth advantage is coincident with loss of the serum-dependency of AHR expression. These results provide a quantitative link between changes in receptor expression and a downstream response, establishing a foundation for future studies of receptor expression and sensitivity to toxic responses in vitro and in vivo.


Potential of mean force between charged colloids: effect of dielectric discontinuities.
The potential of mean force between two spherical and like-charged macroions in a salt-free aqueous solution has been determined using an extended primitive model and canonical Monte Carlo simulations. The systems considered covered the range from a purely repulsive to a purely attractive potential of mean force as the electrostatic coupling was increased. The macroions were modeled as spherical dielectric cavities, and the polarization surface charge densities occurring at the dielectric discontinuities were expanded in spherical harmonics. The surface polarization gave rise to (i) an attenuation of the counterion accumulation at the macroion surfaces at all cases considered, (ii) an enhanced repulsive potential of mean force in the weak to intermediate electrostatic coupling regime, and (iii) a less attractive at short separation and an enhanced attractive potential of mean force at longer macroion-macroion separation in the strong electrostatic coupling regime.


SLC26A4 targeted to the endolymphatic sac rescues hearing and balance in Slc26a4 mutant mice.
Mutations of SLC26A4 are a common cause of human hearing loss associated with enlargement of the vestibular aqueduct. SLC26A4 encodes pendrin, an anion exchanger expressed in a variety of epithelial cells in the cochlea, the vestibular labyrinth and the endolymphatic sac. Slc26a4 (Δ/Δ) mice are devoid of pendrin and develop a severe enlargement of the membranous labyrinth, fail to acquire hearing and balance, and thereby provide a model for the human phenotype. Here, we generated a transgenic mouse line that expresses human SLC26A4 controlled by the promoter of ATP6V1B1. Crossing this transgene into the Slc26a4 (Δ/Δ) line restored protein expression of pendrin in the endolymphatic sac without inducing detectable expression in the cochlea or the vestibular sensory organs. The transgene prevented abnormal enlargement of the membranous labyrinth, restored a normal endocochlear potential, normal pH gradients between endolymph and perilymph in the cochlea, normal otoconia formation in the vestibular labyrinth and normal sensory functions of hearing and balance. Our study demonstrates that restoration of pendrin to the endolymphatic sac is sufficient to restore normal inner ear function. This finding in conjunction with our previous report that pendrin expression is required for embryonic development but not for the maintenance of hearing opens the prospect that a spatially and temporally limited therapy will restore normal hearing in human patients carrying a variety of mutations of SLC26A4.