Activation of alpha2-adrenergic receptors into the lateral parabrachial nucleus enhances NaCl intake in rats.
Water and NaCl intake is strongly inhibited by the activation of alpha(2)-adrenergic receptors with clonidine or moxonidine (alpha(2)-adrenergic/imidazoline agonists) injected peripherally or into the forebrain and by serotonin and cholecystokinin in the lateral parabrachial nucleus (LPBN). Considering that alpha(2)-adrenergic receptors exist in the LPBN and the similar origin of serotonergic and adrenergic afferent pathways to the LPBN, in this study we investigated the effects of bilateral injections of moxonidine alone or combined with RX 821002 (alpha(2)-adrenergic antagonist) into the LPBN on 1.8% NaCl and water intake induced by the treatment with s.c. furosemide (10mg/kg)+captopril (5 mg/kg). Additionally, we investigated if moxonidine into the LPBN would modify furosemide+captopril-induced c-fos expression in the forebrain. Male Holtzman rats with cannulas implanted bilaterally in the LPBN were used. Contrary to forebrain injections, bilateral LPBN injections of moxonidine (0.1, 0.5 and 1 nmol/0.2 microl) strongly increased furosemide+captopril-induced 1.8% NaCl intake (16.6+/-2.7, 44.5+/-3.2 and 44.5+/-4.3 ml/2 h, respectively, vs. vehicle: 6.9+/-1.5 ml/2 h). Only the high dose of moxonidine increased water intake (23.3+/-3.8 ml/2 h, vs. vehicle: 12.1+/-2.6 ml/2 h). Prior injections of RX 821002 (10 and 20 nmol/0.2 microl) abolished the effect of moxonidine (0.5 nmol) on 1.8% NaCl intake. Moxonidine into the LPBN did not modify furosemide+captopril-induced c-fos expression in forebrain areas related to the control of fluid-electrolyte balance. The results show that the activation of LPBN alpha(2)-adrenergic receptors enhances furosemide+captopril-induced 1.8% NaCl and water intake. This enhancement was not related to prior alteration in the activity of forebrain areas as suggested by c-fos expression. Previous and present results indicate opposite roles for alpha(2)-adrenergic receptors in the control of sodium and water intake according to their distribution in the rat brain.


Homocysteine and renal disease.
Hyperhomocysteinemia refers to an elevated circulating level of the sulfur-containing amino acid homocysteine and has been shown to be a risk factor for vascular disease in the general population. In patients with renal failure, hyperhomocysteinemia is a common feature. The underlying pathophysiological mechanism for this phenomenon is unknown. Proposed mechanisms include reduced renal elimination of homocysteine and impaired nonrenal disposal, possibly because of inhibition of crucial enzymes in the methionine-homocysteine metabolism by the uremic milieu. Absolute or relative deficiencies of folate, vitamin B6, or vitamin B12 may also play a role. Several case-control and prospective studies have now indicated that hyperhomocystenemia is an independent risk factor for atherothrombotic disease in patients with predialysis and end-stage renal disease. In renal patients, plasma homocysteine concentration can be reduced by administration of folic acid in doses ranging from 1 to 15 mg per day. In more than 50% of the cases, however, the homocysteine concentration remains above 15 micromol/L. The effects of vitamin B12 or vitamin B6 are unclear. Large intervention trials are now needed to establish whether homocysteine-lowering therapy will reduce atherothrombotic events in patients with renal failure. These studies are now planned or are ongoing.


Curcumin induces apoptosis through mitochondrial pathway and caspases activation in human melanoma cells.
Melanoma is the most malignant skin cancer and is highly resistant to chemotherapy and radiotherapy. Curcumin is a component of turmeric, the yellow spice derived from the rhizome of Curcuma longa. It has been demonstrated to modulate multiple cell signaling pathways, including apoptosis, proliferation, angiogenesis and inflammation. In this study, we studied the signaling pathways involved in melanoma cell death after treatment with curcumin using western blotting. Colorimetric assays (MTT) assessed cell viability. Flow cytometry and DNA laddering evaluated cell apoptosis. Fluorescent microscopy was used to evaluate of Hoechst 33342 staining of nuclei. The result demonstrated that curcumin could induce apoptosis and inhibit proliferation in melanoma cells. Curcumin stimulated the expression of pro-apoptotic Bax, and inhibited the activation of anti-apoptotic Mcl-1 and Bcl-2. During curcumin treatment, caspase-8 and Caspase-3 were cleaved in time and dose-dependent manners. Curcumin treatment also altered the expressions of apoptosis associated proteins NF-κB, p38 and p53. Curcumin induced DNA double strand breaks, which were indicated by phosphorylated H2AX. Our data suggested that curcumin could be used as a novel and effective approach for the treatment of melanoma.


Inhibition of cancer cell proliferation and prostaglandin E2 synthesis by Scutellaria baicalensis.
Scutellaria baicalensis is a widely used Chinese herbal medicine that has been used historically in anti-inflammatory and anticancer therapy. The purpose of this study is to verify its anticancer activity on head and neck squamous cell carcinoma (HNSCC) in vitro and in vivo and to investigate its effect on cyclooxygenase-2 (COX-2), which converts arachidonic acid to prostaglandin E(2) (PGE(2)) and is highly expressed in HNSCC. Two human HNSCC cell lines (SCC-25 and KB) and a nontumorigenic cell line (HaCaT) were tested in vitro for growth inhibition, proliferation cell nuclear antigen expression, and COX-2 activity and expression after treatment with Scutellaria baicalensis extract. Its effects were compared with those of baicalein (a flavonoid isolated from Scutellaria baicalensis), indomethacin (a nonselective COX inhibitor), and celecoxib (a selective COX-2 inhibitor). Four nude mice with s.c. inoculation of KB cells were tested for its anticancer activity in vivo by oral administration of Scutellaria baicalensis at a dose of 1.5 mg/mouse (75 mg/kg), five times/week for 7 weeks. Scutellaria baicalensis and other agents demonstrated a strong growth inhibition in both tested human HNSCC cell lines. No growth inhibition of HaCaT cells was observed with Scutellaria baicalensis. The IC(50)s were 150 micro g/ml for Scutellaria baicalensis, 25 micro M for celecoxib, and 75 micro M for baicalein and indomethacin. Scutellaria baicalensis, as well as celecoxib and indomethacin, but not baicalein, suppressed proliferation cell nuclear antigen expression and PGE(2) synthesis in both cell types. Scutellaria baicalensis inhibited COX-2 expression, whereas celecoxib inhibited COX-2 activity directly. A 66% reduction in tumor mass was observed in the nude mice. Scutellaria baicalensis selectively and effectively inhibits cancer cell growth in vitro and in vivo and can be an effective chemotherapeutic agent for HNSCC. Inhibition of PGE(2) synthesis via suppression of COX-2 expression may be responsible for its anticancer activity. Differences in biological effects of Scutellaria baicalensis compared with baicalein suggest the synergistic effects among components in Scutellaria baicalensis.


Impact of GLP-1 analogue on oxidative damage and hepatic regeneration in experimental 70% hepatectomy model.
The purpose of our study is researching into impact of glucagon like peptide 1 (GLP 1) analogue on liver regeneration after major hepatectomy. 24 wistar albino rats were consecutively divided into 3 groups. Group 1: Control (sham) group day 14 (n = 8), Group 2: Liver resection group day 14 (n = 8); 70% Liver resection was performed, Group 3: Study group day 14 (n = 8); Subsequent to performing 70% liver resection, GLP-1 analogue was administered 2 times a day. (10 μgr/70 kg x 2 times). After 14 day, rats were sacrificed. Oxi- dative stress and antioxidant enzymes and mitochon- drial permeability transition, cytochrome-c, Bax, Bcl- 2, caspase-3, caspase-8 and caspase-3 activity were examined. 70% Liver resection induced oxi- dative stress of liver tissue was ameliorated by GLP-1 induction. Administration of GLP increased Bcl-2 ex- pression. Decreased expression of cytochrome-c was accompanied by a decrease caspase-3, caspase-8, and Bax expression and caspase-3 activity. Glp 1 induction plays a regenerative role in the major hepatectomy. This effect is dependent on modulation of the antiapoptotic and antioxidative pathways by GLP 1 expression.


Microspectroscopic imaging tracks the intracellular processing of a signal transduction protein: fluorescent-labeled protein kinase C beta I.
We have devised a microspectroscopic strategy for assessing the intracellular (re)distribution and the integrity of the primary structure of proteins involved in signal transduction. The purified proteins are fluorescent-labeled in vitro and reintroduced into the living cell. The localization and molecular state of fluorescent-labeled protein kinase C beta I isozyme were assessed by a combination of quantitative confocal laser scanning microscopy, fluorescence lifetime imaging microscopy, and novel determinations of fluorescence resonance energy transfer based on photobleaching digital imaging microscopy. The intensity and fluorescence resonance energy transfer efficiency images demonstrate the rapid nuclear translocation and ensuing fragmentation of protein kinase C beta I in BALB/c3T3 fibroblasts upon phorbol ester stimulation, and suggest distinct, compartmentalized roles for the regulatory and catalytic fragments.


Inhibition of interleukin 10 by rituximab results in down-regulation of bcl-2 and sensitization of B-cell non-Hodgkin's lymphoma to apoptosis.
Treatment of patients with non-Hodgkin's lymphoma (NHL) is frequently hampered by development of chemoresistance. Rituximab is a chimeric mouse antihuman CD20 antibody that offers an alternative; however, its mechanism of action is not clearly understood. Treatment of lymphoma cell lines with Rituximab sensitizes the cells to the cytotoxic and apoptotic effects of therapeutic drugs, e.g., cisplatin, fludarabine, vinblastine, and Adriamycin. This study investigated the mechanism(s) involved in the reversal of drug resistance by Rituximab therapy. NHL cells synthesize and secrete antiapoptotic cytokines implicated in drug resistance, including interleukin (IL)-6, IL-10, and tumor necrosis factor alpha. We hypothesized, therefore, that sensitization by Rituximab may be due in part to modification of cytokine production. In this study, examination of cytokine secretion by NHL 2F7 tumor cells revealed down-regulation of IL-10 by Rituximab treatment. Moreover, cytotoxicity assays using exogenous IL-10 and IL-10-neutralizing antibodies demonstrated that IL-10 serves as an antiapoptotic/protective factor in these tumor cells against cytotoxic drugs. Furthermore, expression in 2F7 cells of the protective factor, Bcl-2, was shown to be dependent on IL-10 levels and down-regulated by Rituximab. Other gene products such as Bax, Bcl-x, Bad, p53, c-myc, and latent membrane protein-1 (LMP) were not affected by Rituximab treatment. Drug sensitization, as well as down-regulation of both IL-10 and Bcl-2, was corroborated in experiments using the NHL cell line 10C9. The Ramos and Daudi NHL cell lines were not sensitizable, nor did their Bcl-2 or IL-10 levels change. These studies demonstrate that one mechanism by which Rituximab sensitizes NHL to chemotherapeutic drugs is mediated through down-regulation of antiapoptotic IL-10 autocrine/paracrine loops and Bcl-2. The clinical relevance of these findings is discussed.


Is Community-engaged Learning Possible During a Pandemic: A Call for Culturally Competent Medical Education.
This article was migrated. The article was marked as recommended. Previously, medical school curriculum focused on faculty or physician-led basic science and traditional clinical experiences, with medical students only gaining experience of the community in which they practice during residency. In an effort to enable students to understand US healthcare disparities, the introduction of public health topics regarding marginalized communities and underrepresented individuals have been included in the classroom. However, missing from this shift is the inclusion of authentic public health educational experiences for medical students. These learning experiences are vital to truly understanding the marginalized and discriminated patient populations physicians will encounter. The recent COVID-19 pandemic has brought forth challenges for medical educators in numerous ways including how to effectively prepare students in understanding cultural competency through community-engaged learning for a new set of patient population; the pandemic patient. Due to health disparities, each patient experienced this pandemic differently based on their individual, cultural and community setting; also highlighting the importance of community-engaged learning. Here, the authors posit the role and importance of community-engaged learning in medical education and its utilization during the changing medical landscape due to COVID-19.


Bariatric surgery and bone disease: from clinical perspective to molecular insights.
The use of bariatric surgery for the treatment of morbid obesity has increased annually for the last decade. Although many studies have demonstrated the efficacy and durability of bariatric surgery for weight loss, there are limited data regarding long-term side effects of these procedures. Recently, there has been an increased focus on the impact of bariatric surgery on bone metabolism. Bariatric surgery utilizes one or more of three mechanisms of action resulting in sustained weight loss. These include restriction (gastric banding, vertical banded gastroplasty and sleeve gastrectomy), malabsorption surgery with or without associated restriction (Roux en Y gastric bypass, duodenal switch, biliopancreatic diversion and jejunoileal bypass) and changes in gut-derived hormones that control energy metabolism also referred to as neuro-hormonal control of energy metabolism (Roux en Y gastric bypass, duodenal switch, biliopancreatic diversion, jejunoileal bypass, surgical procedures as above and gastric sleeve). Weight reduction has been associated with increased bone resorption but the mechanisms behind this have not yet been fully elucidated. Each of the mechanisms of action of bariatric surgery (restriction, malabsorption, neuro-hormonal control of energy metabolism) may uniquely affect bone resorption. In this paper we will review the current state of knowledge regarding the relationship between bariatric surgery and bone metabolism with emphasis on possible mechanisms of action such as malnutrition, hormonal interactions and mechanical unloading of the skeleton. Further, we suggest a future research agenda.


Global public health leadership for the twenty-first century: towards improved health of all populations.
We are at an unprecedented moment in history in terms of the health of populations around the world. New and old problems all require both short- and long-term solutions, at the individual, community, national and global levels. Unique solutions for each challenge may not be feasible or adequately effective or cost-effective. We are confronted by health systems that are not well matched to current and future needs, both for sustained prevention and chronic care. Moving forward effectively as a field will benefit from a focus on the changing needs of global health, and on how changing conditions, globally, should define the next generation of public health leadership so as to best accomplish global health goals.


The architecture of variant surface glycoprotein gene expression sites in Trypanosoma brucei.
Trypanosoma brucei evades the immune system by switching between Variant Surface Glycoprotein (VSG) genes. The active VSG gene is transcribed in one of approximately 20 telomeric expression sites (ESs). It has been postulated that ES polymorphism plays a role in host adaptation. To gain more insight into ES architecture, we have determined the complete sequence of Bacterial Artificial Chromosomes (BACs) containing DNA from three ESs and their flanking regions. There was variation in the order and number of ES-associated genes (ESAGs). ESAGs 6 and 7, encoding transferrin receptor subunits, are the only ESAGs with functional copies in every ES that has been sequenced until now. A BAC clone containing the VO2 ES sequences comprised approximately half of a 330 kb 'intermediate' chromosome. The extensive similarity between this intermediate chromosome and the left telomere of T. brucei 927 chromosome I, suggests that this previously uncharacterised intermediate size class of chromosomes could have arisen from breakage of megabase chromosomes. Unexpected conservation of sequences, including pseudogenes, indicates that the multiple ESs could have arisen through a relatively recent amplification of a single ES.


Epidemiology of foodborne Norovirus outbreak in Incheon, Korea.
On June 14, 2008, an outbreak of gastroenteritis occurred among elementary school students in Incheon. We conducted an investigation to identify the source and described the extent of the outbreak. We performed a retrospective cohort study among students, teachers and food handlers exposed to canteen food in the elementary school. Using self-administered questionnaires we collected information on symptoms, days of canteen food eaten, food items consumed. Stool samples were collected from 131 symptomatic people and 11 food handlers. The catering kitchen was inspected and food samples were taken. Of the 1,560 people who ate canteen food, 117 were symptomatic cases, and the attack rate was 7.5%. Consumption of cucumber-crown daisy salad (RR=2.71), fresh cabbage mix (RR=2.23), dried radish salad (RR=3.04) and young radish kimchi (RR=2.52) were associated with illness. Sixty-four (45%) of the 142 stool specimens were positive for Norovirus. Norovirus was detected in 2 food handlers. Interviews with kitchen staff indicated the likelihood of contamination from an infected food handler to the dried radish salad during food processing. The excretion of Norovirus from asymptomatic food handlers may be an infection source of Norovirus outbreaks.


Whole Slide Imaging Versus Microscopy for Primary Diagnosis in Surgical Pathology: A Multicenter Blinded Randomized Noninferiority Study of 1992 Cases (Pivotal Study).
Most prior studies of primary diagnosis in surgical pathology using whole slide imaging (WSI) versus microscopy have focused on specific organ systems or included relatively few cases. The objective of this study was to demonstrate that WSI is noninferior to microscopy for primary diagnosis in surgical pathology. A blinded randomized noninferiority study was conducted across the entire range of surgical pathology cases (biopsies and resections, including hematoxylin and eosin, immunohistochemistry, and special stains) from 4 institutions using the original sign-out diagnosis (baseline diagnosis) as the reference standard. Cases were scanned, converted to WSI and randomized. Sixteen pathologists interpreted cases by microscopy or WSI, followed by a wash-out period of ≥4 weeks, after which cases were read by the same observers using the other modality. Major discordances were identified by an adjudication panel, and the differences between major discordance rates for both microscopy (against the reference standard) and WSI (against the reference standard) were calculated. A total of 1992 cases were included, resulting in 15,925 reads. The major discordance rate with the reference standard diagnosis was 4.9% for WSI and 4.6% for microscopy. The difference between major discordance rates for microscopy and WSI was 0.4% (95% confidence interval, -0.30% to 1.01%). The difference in major discordance rates for WSI and microscopy was highest in endocrine pathology (1.8%), neoplastic kidney pathology (1.5%), urinary bladder pathology (1.3%), and gynecologic pathology (1.2%). Detailed analysis of these cases revealed no instances where interpretation by WSI was consistently inaccurate compared with microscopy for multiple observers. We conclude that WSI is noninferior to microscopy for primary diagnosis in surgical pathology, including biopsies and resections stained with hematoxylin and eosin, immunohistochemistry and special stains. This conclusion is valid across a wide variety of organ systems and specimen types.


Diagnosis and differential diagnosis of fibromyalgia.
Fibromyalgia is a chronic functional illness that presents with widespread musculoskeletal pain as well as a constellation of symptoms including fatigue, cognitive dysfunction, sleep difficulties, stiffness, anxiety, and depressed mood. The diagnosis of fibromyalgia, similar to other functional disorders, requires that organic diseases are not causing the symptoms. Systemic and rheumatic diseases can be ruled out by a patient history, physical examination, and laboratory investigations. Because there are no specific laboratory tests for fibromyalgia, the 1990 American College of Rheumatology (ACR) classification criteria have been used in clinical settings; however, they are not ideal for individual patient diagnosis. Clinicians should be aware of limitations inherent in using tender points in the diagnosis of fibromyalgia. The multiple symptoms of fibromyalgia often overlap with those of related disorders and may further complicate the diagnosis. One of the most challenging diagnostic dilemmas that clinicians face is distinguishing fibromyalgia from other central pain disorders (e.g., irritable bowel syndrome, chronic fatigue syndrome, migraine). Screening questions based on published criteria can be used as a first approach in diagnosing functional illnesses. Numerous studies report a higher prevalence of psychiatric disorders in patients with fibromyalgia. Therefore, a careful history and evaluation should be taken for the presence of primary mood disturbances. To date, there is no "gold standard" for diagnosing fibromyalgia. Until a better clinical case definition of fibromyalgia exists, all diagnostic criteria should be interpreted with caution, considered rudimentary, and subject to modification.


Increased levels of homocysteine in patients with ulcerative colitis.
To investigate serum levels of homocysteine (Hcys) and the risk that altered levels carry for thrombosis development in ulcerative colitis (UC) patients. 55 UC patients and 45 healthy adults were included. Hcys, vitamin B12 and folic acid levels were measured in both groups. Clinical history and thromboembolic events were investigated. The average Hcys level in the UC patients was 13.3 +/- 1.93 micromol/L (range 4.60-87) and was higher than the average Hcys level of the control group which was 11.2 +/- 3.58 micromol/L (range 4.00-20.8) (P < 0.001). Vitamin B12 and folic acid average values were also lower in the UC group (P < 0.001). When multivariate regression analysis was performed, it was seen that folic acid deficiency was the only risk factor for hyperhomocysteinemia. Frequencies of thromboembolic complications were not statistically significantly different in UC and control groups. When those with and without a thrombosis history in the UC group were compared according to Hcys levels, it was seen that there were no statistically significant differences. A negative linear relationship was found between folic acid levels and Hcys. We could not find any correlations between Hcys levels and history of prior thromboembolic events.


Evidence for molecular differences in prostate cancer between African American and Caucasian men.
The aim of this study was to compare the frequency of ERG rearrangement, PTEN deletion, SPINK1 overexpression, and SPOP mutation in prostate cancer in African American and Caucasian men. Dominant tumor nodules from radical prostatectomy specimens of 105 African American men (AAM) were compared with 113 dominant nodules from Caucasian men (CaM). Clinical and pathologic characteristics of the two groups were similar. SPINK1 overexpression was evaluated by immunohistochemistry, ERG rearrangement and PTEN deletion by FISH, and SPOP mutation by Sanger sequencing. ERG rearrangement was identified in 48 of 113 tumors (42.5%) in CaM and 29 of 105 tumors (27.6%) in AAM (P = 0.024). PTEN deletion was seen in 19 of 96 tumors (19.8%) in CaM and 7 of 101 tumors (6.9%) in AAM (P = 0.011). SPINK1 overexpression was present in 9 of 110 tumors (8.2%) in CaM and 25 of 105 tumors (23.4%) in AAM (P = 0.002). SPOP mutation was identified in 8 of 78 (10.3%) tumors in CaM and 4 of 88 (4.5%) tumors in AAM (P = 0.230). When adjusted for age, body mass index, Gleason score, and pathologic stage, ERG rearrangement and SPINK1 overexpression remain significantly different (P = 0.018 and P = 0.008, respectively), and differences in PTEN deletion and SPOP mutation approach significance (P = 0.061 and P = 0.087, respectively). Significant molecular differences exist between prostate cancers in AAM and CaM. SPINK1 overexpression, an alteration associated with more aggressive prostate cancers, was more frequent in AAM, whereas ERG rearrangement and PTEN deletion were less frequent in this cohort. Further investigation is warranted to determine whether these molecular differences explain some of the disparity in incidence and mortality between these two ethnic groups.


Urinary and Faecal Incontinence: Point Prevalence and Predictors in a University Hospital.
Incontinence is common and associated with adverse outcomes. There are insufficient point prevalence data for incontinence in hospitals. We evaluated the prevalence of urinary (UI) and faecal incontinence (FI) and their predictors among inpatients in an acute university hospital on a single day. Continence status was recorded using the modified Barthel Index (BI). Baseline characteristics, Clinical Frailty Scale (CFS) and ward type were recorded. In all, 435 patients were assessed, median age 72 ± 23 years and 53% were male. The median CFS score was 5 ± 3. The point prevalence of UI was 26% versus 11% for FI. While UI and FI increased with age, to 35.2% and 21.1% respectively for those ≥85, age was not an independent predictor. Incontinence also increased with frailty; CFS scores were independently associated with both UI (p = 0.006) and FI (p = 0.03), though baseline continence status was the strongest predictor. Patients on orthopaedic wards had the highest prevalence of incontinence. Continence assessments were available for only 11 (2%) patients. UI and FI are common conditions affecting inpatients; point prevalence increases with age and frailty status. Despite this, few patients receive comprehensive continence assessments. More awareness of its high prevalence is required to ensure incontinence is adequately managed in hospitals.


Heterogeneity of bile pigment conjugates as revealed by chromatography of their ethyl anthranilate azopigments.
1. Azopigments derived from conjugated bile pigments by coupling with the diazonium salt of ethyl anthranilate are analysed conveniently by quantitative t.l.c. or by column chromatography on CM-cellulose. 2. By chromatographic studies combined with a series of chemical tests six groups of azopigments were demonstrable in preparations from bile and from icteric urine of man. Azobilirubin and its beta-d-monoglucuronide have hitherto been considered to be the only major derivatives that can be obtained from human bile pigments. In the present work, other azopigments accounted for 30-40% of the total azopigment material, and the amounts of these showed considerable variation among biological fluids. 3. The divergence of the present results from earlier work is probably related to the use of milder diazotization conditions and of chromatographic techniques with a high resolving power. 4. The thin-layer chromatographic systems developed allow rapid and quantitative analysis of azopigments derived from bile pigments.


Low-grade inflammation differentiates between symptoms of apathy and depression in community-dwelling older individuals.
Systemic low-grade inflammation has repeatedly been associated with depression in old age, but the relationship with apathy is less clear. The present study assessed whether C-reactive protein (CRP) is differentially associated with symptoms of apathy and depression. A population-based cohort study was carried-out. At baseline and after two and four years of follow-up, CRP levels were assessed and symptoms of apathy and depression were measured using the 15-item Geriatric Depression Scale. Logistic regression analysis was used to investigate the cross-sectional and longitudinal associations of CRP with symptoms of apathy and depression. Two thousand forty-seven community-dwelling participants (70-78 years) without a history of cardiovascular disease or stroke were studied. A cross-sectional association was found between CRP and apathy symptoms at three time points (odds ratio (OR) per natural log unit increase in CRP: baseline visit = 1.40, 95% CI = 1.12-1.75; two-year follow-up visit = 1.62, 95% CI = 1.17-2.25; four-year follow-up visit = 1.51, 95% CI = 1.03-2.21). This did not change after adjustment for demographics and depressive symptoms, and was slightly attenuated after adjustment for cardiovascular risk factors. No cross-sectional association was found with depressive symptoms. Baseline CRP did not predict incident apathy or depressive symptoms during four years of follow-up. Increased CRP levels are associated with apathy symptoms but not with depressive symptoms. This suggests a differential effect of inflammation on apathy and depression. In older persons, symptoms of apathy may be a behavioral manifestation of concurrent low-grade inflammation.


Investigation of outbreaks of salmonella in hospitals.
In a two year prospective survey of outbreaks of salmonella infection in hospitals in England and Wales 55 outbreaks were identified. Reports of investigation of these outbreaks were reviewed for evidence of food borne infection and cross infection. Food borne infection probably accounted for only six outbreaks, but these made up 40% of the 15 outbreaks in which there were more than five patients and staff with symptoms. Person to person transmission was the probable mode of spread in most outbreaks. It is recommended that in addition to bacteriological investigations clinical and epidemiological data should be collected to implicate food or other possible common vehicles of infection. Bacteriological screening of patients and staff who do not have symptoms may be unrewarding in the absence of epidemiological data to define groups at risk of infection.


Diet drug-related cardiac valve disease: the Mayo Clinic echocardiographic laboratory experience.
To describe the prevalence of diet drug-related valvular disease among our referral population and the association of valvular disease with duration of exposure to fenfluramine and phentermine in combination and to dexfenfluramine alone. In this retrospective review of clinical and echocardiographic data, charts of patients referred for treatment of toxic effects of diet drugs were reviewed, and telephone interviews were conducted. Between June and December 1997, 191 patients (164 women, 27 men; mean age, 47 years) were referred for possible diet drug-related valvular disease. Twenty-eight (28%) of the 99 asymptomatic patients and 40 (43%) of the 92 symptomatic patients had abnormal echocardiographic findings. Valvular lesions among the 68 patients with abnormal echocardiographic findings included mild (or greater) aortic regurgitation in 55 patients (81%), moderate (or greater) mitral regurgitation in 12 (18%), and moderate (or greater) tricuspid regurgitation in 7 (10%). The Food and Drug Administration case definition of diet drug-related valvulopathy was noted in 31 % of this referral population. Of patients with valvulopathy, mean duration of therapy with fenfluramine and phentermine in combination and dexfenfluramine alone was 9 months and 5 months, respectively. Duration of therapy was not associated with presence or absence of disease. Five patients had surgical intervention for severe valvulopathy: 3 had mitral valve repair, 1 had mitral valve replacement, and 1 had aortic valve replacement. Pulmonary hypertension (>40 mm Hg) was found in 24 patients (13%), and 17 (71 %) had pulmonary hypertension in association with valvulopathy. This study demonstrated a 31% (60/191) prevalence of valvulopathy in patients with a history of diet drug exposure who were referred for echocardiographic evaluation. The most common finding was mild aortic regurgitation. Twenty-eight percent of asymptomatic patients had abnormal echocardiographic findings. This study emphasizes the spectrum of diet drug-related cardiac disease and the potential for valvulopathy in asymptomatic patients.


Dose-response relation of diazoxide in children with hypertension.
Diazoxide was administered to sixteen pediatric patients (ages 10 months to 13 years) with secondary forms of hypertension. Admission BP was 178+/-8/130+/-5 mm Hg (mean +/- SEM). Diazoxide was administered rapidly intravenously in doses ranging from 2 to 7.5 mg/kg. A significant (P less than 0.001), linear log dose-response relation was obtained which showed that a 3 mg/kg dose of diazoxide lowered diastolic BP by an average of 30 mm Hg. In five patients reduction of idastolic BP by a single injection of diazoxide was no different than when the same total dose was given as two or three small injections repeated at fifteen to twenty minute intervals. It is concluded that 1) many hypertensive children respons significantly to doses of diazoxide smaller than the usually recommended 5 mg/kg; 2) diazoxide has a significant dose-response relation in hypertensive pediatric patients; and 3) the desired blood pressure response in hypertensive children can be titrated using repeated small injections of diazoxide.


Fludarabine and cytarabine as continuous sequential infusion for elderly patients with acute myeloid leukemia.
A phase II study was conducted to investigate the effects of a therapeutic program based on the combination of fludarabine and cytarabine (ARA-C) administered as a sequential continuous infusion in untreated elderly patients with acute myeloid leukemia (AML). Sixty-three patients with non-M3 AML, median age 69 years (range 61-81), were accrued. Twenty-four patients (38%) had AML secondary to myelodysplastic syndrome. Fludarabine and ARA-C were administered as a continuous sequential infusion for 72 and 96 hours, respectively, after a loading dose. Patients achieving complete remission (CR) were intended to receive an additional course, followed by autologous stem cell transplantation (ASCT). Overall, 42 patients (67%) achieved CR. There were 10 induction deaths (16%), while 11 patients were refractory (17%). Among those achieving a remission, 35 patients (83%) received the planned consolidation course and 29 underwent mobilization of CD34+ cells into the peripheral blood for collection, which was successful in 23 (79%). Overall, 17 patients (27% of the whole population) received ASCT. The median overall and disease-free survival were both 10 months. Patients with an intermediate karyotype and those receiving ASCT had a significantly better clinical outcome. Results in terms of CR achievement, CD34+ cell collection and ASCT feasibility. A longer follow up is needed in order to evaluate the actual benefit on long-term survival.


[Various aspects of the use of liposomes in the diagnosis, prophylaxis and treatment of infectious diseases].
The literature data on preparation and properties of liposomes containing drugs, antigens and immunostimulants are reviewed. The efficiency of antibiotic-containing liposomal preparations in the treatment of infectious diseases has been demonstrated due to the directed transport of drugs into the target organs, inside cells, and to the decrease of antibiotics toxic effects. Immunostimulating and adjuvant effects of antigen-containing liposomes, particularly in combination with immunostimulants, have been described, that permits obtaining highly active diagnostic sera. The data are presented on the prospects for construction of molecular vaccines with the use of liposomes.


Rapid Notch1 nuclear translocation after ligand binding depends on presenilin-associated gamma-secretase activity.
Recent data suggest an intimate relationship between the familial Alzheimer disease gene presenilin 1 (PS1) and proteolytic processing of both the amyloid precursor protein (APP) and the important cell signaling molecule, Notch1. We now show, using mammalian cells transfected with full-length Notch1, that the C terminal domain of Notch1 rapidly translocates to the nucleus upon stimulation with the physiologic ligand Delta and initiates a CBF1-dependent signal transduction cascade. Using this assay, we demonstrate that the same aspartate mutations in PS1 that block APP processing also prevent Notch1 cleavage and translocation to the nucleus. Moreover, we show that two APP gamma-secretase inhibitors also diminish Notch1 nuclear translocation in a dose-dependent fashion. However, Notch1 signaling, assessed by measuring the activity of CBF1, a downstream gene, was reduced but not completely abolished in the presence of either aspartate mutations or gamma-secretase inhibitors. Our results support the hypothesis that similar PS1-related enzymatic activity is necessary for both APP and Notch1 processing, yet suggest that Notch signaling may remain relatively preserved with moderate levels of gamma-secretase inhibition.


Further observations on the inhibitory effect of extracellular potassium ions on glycine uptake by mouse ascites-tumour cells.
1. The initial rate of uptake of glycine by the tumour cells was measured as a function of the Na(+) and K(+) concentrations in the solution in which the cells were suspended. When [Gly] was 1mm or 12mm, the rate in the absence of Na(+) was independent of [K(+)] and about 3% or 10% respectively of the rate when [Na(+)] was 150m-equiv./l. 2. The Na(+)-dependent glycine entry rate, v, at a given value of [Na(+)] was successively lowered when [K(+)] was increased from 8 to 47 to 96m-equiv./l. A kinetic analysis indicated that K(+) competitively inhibited the action of Na(+). The results were in fair agreement with previous determinations of the kinetic parameters. 3. The presence of 2mm-sodium cyanide and 10mm-2-deoxyglucose lowered the cellular ATP content to less than 3% of the value in the respiring cells. Although v was then about 50% smaller, the relative effects of K(+) and Na(+) on the system were similar to those observed during respiration. 4. A theoretical analysis indicated that the variation of v with [K(+)] is not a reliable guide to the extent to which the K(+) gradient between the cells and their environment may contribute to the net transport of glycine.


Design aspects of vaginal applicators that influence acceptance among target users.
Although sensory-guided product design is most traditionally used by food and beverage companies, the approach has widespread application for many other products, including pharmaceuticals and medical devices. Previously, our team used sensory methods to explore preclinical optimization of soft-gel vaginal microbicides. Past clinical trials suggest vaginal microbicides may be an effective means for women to protect themselves from HIV and other sexually transmitted infections, but these microbicides will not work if they are not used due to poor acceptability. Our prior work suggests properties like firmness, size, and shape all influence women's willingness to try soft-gel vaginal suppositories. As product insertion is part of the overall experience of using vaginal microbicides, understanding the features of vaginal applicators that appeal to women, and incorporating these insights into vaginal drug delivery systems, may also improve user adherence. Despite widespread use of vaginal applicators, there is minimal public data on women's perceptions of and preferences for physical applicator features. Other work suggests women want vaginal applicators that are single use, pre-filled, made of plastic, and easy to use, store, and discard. Applicator attributes that may be important to women, such as length, color, or visual appeal, have not been investigated previously. The objective of this research was to understand what physical applicator attributes are appealing to women. Here, 18 commercially available applicators were evaluated by a convenience sample of women (n = 102) for overall liking and perceptions of various attributes (perceived length and width, ease-of-grip, expected ease-of-use, expected comfort inside the body, visual appeal, color liking, and environmental friendliness). Preference mapping using both liking data and attribute data showed attributes such as color, visual appeal, ease of grip, expected ease of use, and expected comfort inside the body drove higher liking ratings for applicators, while perceived length negatively affected liking. In general, plastic tampon applicators contained more positive features and were better liked relative to a cardboard tampon applicator or applicators for insertion of medicated gels or suppositories. Incorporating more desirable features into applicators meant for insertion of vaginal microbicides or other vaginal medications may improve the user experience, and possibly user adherence.


Traumatic carotid artery dissection caused by bungee jumping.
Bungee jumping is a popular recreational activity in which participant experiences transient freefall while connected to a bungee cord. The rapid freefall and the resultant rebound force created by the bungee cord can result in a variety of bodily injuries. We report herein a case of traumatic carotid artery dissection caused by bungee jumping. The symptoms related to carotid artery dissection were successfully treated with anticoagulation. The etiology of carotid dissection related to bungee jumping is discussed. Physicians should be cognizant of this potential injury due to the force created by the freefall and rebound motion associated in this recreational sport.


Leukemia markers expression of peripheral blood vs bone marrow blasts using flow cytometry.
Flow cytometric techniques are widely used in clinical hematology. Characterization of leukemias by immunotyping is particularly helpful when the morphology is difficult to interpret. The major advantage of using immune markers by flow cytometry is the identification of particular leukemia subtype, not recognized by morphologic criteria, which may have prognostic significance. Current literature suggests when peripheral blood (PB) is consisted of 30% blasts or higher diagnosis of acute leukemia is most likely. However, bone marrow aspiration may also be performed as a confirmatory diagnosis. Immunotyping of PB and BM in leukemias not only determine the decision making for a specific therapeutic regimen, but also is a practical prognostic indicator. We evaluated 18 patients with acute myeloid Leukemia (AML) and 13 patients with acute lymphoid leukemia (ALL). In all cases, the amount of blasts in PB was 30% or higher. Two ml PB and BM samples from each patient was collected. Following the preparation process, expression of markers was detected by using flow cytometry. The panel of monoclonal antibodies used in this study were consisted of CD3, CD7, CD5 (for T lymphocytes lineage); CD19, CD22, CD20, CD10 (for B lymphocytes lineage); CD13, CD14, CD33 (for myeloid subsets); and TDT, HLA-DR, CD45 (non lineage restricted). Expression levels of PB and BM markers were compared by using statistical analysis. The results showed apparent discrepancies for some markers in ALL group. However, in AML patients most of the selected markers have shown considerable correlation between PB and BM samples. Only four markers (CD13, CD14, CD45, and HLA-DR) showed positive correlation. In contrast, most markers (CD3, CD5, CD13, CD14, CD19, CD45, HLA-DR, and TdT) showed strong correlation between PB and BM samples in AML group. The findings of this study suggests that targeted gating strategy for blast population as well as selection of a suitable panel of monoclonal antibodies may be essential for diagnosis of leukemia resulting in similar immunotyping pattern in PB and BM. Although our results are preliminary, this can minimize the necessity of BM aspiration for leukemia patients.


Well-differentiated Liposarcoma in a Bonnet Macaque (Macaca radiata).
Here we describe the occurrence of a subcutaneous liposarcoma in a geriatric bonnet macaque (Macaca radiata). Clinical presentation was a rapidly growing, ulcerated, subcutaneous mass in the umbilical region of a 28-y-old intact female macaque. The mass was successfully removed through excisional biopsy, and histopathology provided a morphologic diagnosis of well-differentiated liposarcoma. The macaque recovered without complication and displayed no signs of recurrence for at least 18 mo after excision. To our knowledge, this case represents the first report of liposarcoma in a bonnet macaque.


Long-term overweight and weight gain in early adulthood in association with risk of endometrial cancer.
Long-term overweight and substantial weight gain over adulthood are known risk factors of endometrial cancer, but the timing of weight gain in relation to risk and the effect of weight change on age at diagnosis remain unclear. A population-based case-control study was conducted to evaluate the long-term effect of body weight on endometrial cancer risk. The study enrolled 668 incident cases and 674 population controls. Anthropometric features in each decade of adult life were ascertained through in-person interview and analyzed for their associations with endometrial cancer using unconditional logistic regression. As expected, high body mass index (BMI) was significantly associated with increased risk. Women who were overweight or obese at the time of interview had adjusted odds ratios of 1.54 (95%CI 1.13-2.10) and 4.76 (95%CI 3.50-6.49), respectively, compared to women of normal weight. Similar associations were observed for BMI assessed at each decade of adult life. More importantly, women who were overweight (BMI ≥ 25) in their 20s or 30s and maintained the overweight throughout life had significantly higher risk than those who became overweight at ages 40s or 50s. Women with substantial weight gain (≥35%) in early adulthood (age 20s) developed the disease 10 years earlier than those without such weight change in early life. These observations further confirm the critical link between body weight and development of endometrial cancer.


Anti-HCV treatment with ombitasvir/paritaprevir/ritonavir ± dasabuvir is associated with increased bile acid levels and pruritus.
Direct acting antiviral (DAA)-based treatment with ombitasvir/paritaprevir/ritonavir ± dasabuvir (OBV/PTV/r ± DSV) is highly effective in HCV genotype 1 or 4 infection and well-tolerated with only few side effects. However, pruritus has been observed in several trials in up to 20% of patients and seems to be unique for this DAA combination. The aim of this preliminary study was to investigate the effect of OBV/PTV/r ± DSV on bile acid levels and to correlate them to the emergence of pruritus during treatment. Twenty patients with chronic hepatitis C genotype 1 or 4 were treated for 12 or 24 weeks with OBV/PTV/r ± DSV with or without ribavirin. Side effects including pruritus were assessed every 4 weeks during treatment or on demand. Blood was collected in fasting state at baseline and at treatment week 4 for determination of bile acid concentrations by high-resolution mass spectrometry. Pruritus developed in 5 out of 20 patients during the first 4 weeks of DAA treatment. Pruritus was self-limiting during DAA treatment in 4 patients while one patient required cholestyramine treatment and responded well. Total bile acid levels increased approximately 4‑fold by treatment week 4. Pruritus observed during OBV/PTV/r ± DSV treatment of chronic hepatitis C is associated with increased on-treatment serum bile acid levels, possibly due to ritonavir-induced alterations of bile acid transport.


The course of depression 10-weeks post-acute coronary syndrome: assessment using the cardiac depression visual analogue scale.
This study investigated depressed mood post-acute coronary syndrome in an Australian sample using the Cardiac Depression Visual Analogue Scale (CD-VAS), which was developed to assess the depression-related symptoms of reduced pleasure, memory, activity, future pessimism, increased irritation and sleep problems. The Beck Depression Inventory-II, the Cardiac Depression Scale (CDS) and a demographic medical questionnaire were administered at 2 weeks post-ACS to an initial sample of 13 female and 45 male participants whose mean age was 59.16 (SD=11.13) years (range 38-79). Depressed and non-depressed participants, determined by the CDS, then kept a CD-VAS diary between 2 and 12 weeks. 32 participants kept the diary for the entire 10 weeks. Total CD-VAS scores remained elevated in the depressed group for the entire period. The groups differed on the Pleasure, Memory and Future items for a total of 6, 6 and 5 weeks, respectively, and differed on the Activity item only at 1, 3 and 5 weeks. No group differences were seen in the Irritation or Sleep items. Within the depressed group, the only CD-VAS items that differed from each other across time were Irritation and Activity. Further, the Sleep, Irritation and Activity items had a significant time effect. These results indicate that although overall depression, as measured by the CD-VAS, remained elevated in the depressed group, there were key differences in recovery rates of specific depression-related symptoms. These differences could be useful indicators in the treatment of depression post-ACS.


A pharmacogenetic study of trimethylaminuria in Orientals.
A study of the metabolism of trimethylamine was carried out in 103 healthy Thai volunteers (70 men and 33 women) and it was found that under normal dietary conditions 84-100% of trimethylamine was excreted in the urine in its N-oxide form. Five propositi living in different parts of the country were identified as having deficiency in the N-oxidation of this tertiary amine, because they excreted only 8-35% of this chemical as trimethylamine N-oxide. This metabolic defect was also confirmed by the results of an oral trimethylamine (600 mg) challenge experiment in which all five propositi were found to excrete an even smaller percentage of trimethylamine as trimethylamine N-oxide in their urine. The results of a study of the families of the two proband individuals, as well as those members of their preceding generations under normal dietary conditions, are consistent with the view that the disorder or metabolic defect is inherited in a Mendelian fashion as an autosomal recessive trait, similar to that reported for white Caucasian subjects.


Hunger increases delay discounting of food and non-food rewards.
How do our valuation systems change to homeostatically correct undesirable psychological or physiological states, such as those caused by hunger? There is evidence that hunger increases discounting for food rewards, biasing choices towards smaller but sooner food reward over larger but later reward. However, it is not understood how hunger modulates delay discounting for non-food items. We outline and quantitatively evaluate six possible models of how our valuation systems modulate discounting of various commodities in the face of the undesirable state of being hungry. With a repeated-measures design, an experimental hunger manipulation, and quantitative modeling, we find strong evidence that hunger causes large increases in delay discounting for food, with an approximately 25% spillover effect to non-food commodities. The results provide evidence that in the face of hunger, our valuation systems increase discounting for commodities, which cannot achieve a desired state change as well as for those commodities that can. Given that strong delay discounting can cause negative outcomes in many non-food (consumer, investment, medical, or inter-personal) domains, the present findings suggest caution may be necessary when making decisions involving non-food outcomes while hungry.


Porphyrin and tetrabenzoporphyrin dendrimers: tunable membrane-impermeable fluorescent pH nanosensors.
The pH dependencies of the UV-vis and fluorescent spectra of new water-soluble dendritic porphyrins and tetrabenzoporphyrins were studied. Because of extended pi-conjugation and nonplanar distortion, the absorption and the emission bands of tetraaryltetrabenzoporphyrins (Ar(4)TBP) are red-shifted and do not overlap with those of regular tetraarylporphyrins (Ar(4)P). When encapsulated inside dendrimers with hydrophilic outer layers, Ar(4)Ps and Ar(4)TBPs become water soluble and can serve as pH indicators, with pK's adjustable by the peripheral charges on the dendrimers. Two new dendritic porphyrins, Gen 4 polyglutamic porphyrin dendrimer H(2)P-Glu(4)OH (1) with 64 peripheral carboxylates and Gen 1 poly(ester amide) Newkome-type tetrabenzoporphyrin dendrimer H(2)TBP-Nw(1)OH (2) with 36 peripheral carboxylates, were synthesized and characterized. The pK's of the encapsulated porphyrins (pK(H)()2(P)(-)(Glu)()4(OH) = 6.2 and pK(H)()2(TBP)(-)(Nw)()1(OH) = 6.3) were found to be strongly influenced by the dendrimers, revealing significant electrostatic shielding of the cores by the peripheral charges. The titration curves obtained by differential excitation using the mixtures of the dendrimers were shown to be identical to those determined for the dendrimers individually. Due to their peripheral carboxylates and nanometric molecular size, porphyrin dendrimers cannot penetrate through phospholipid membranes. Dendrimer 1 was captured inside phospholipid liposomes, which were suspended in a solution containing dendrimer 2. No response from 1 was detected upon pH changes in the bulk solution, while the response from 2 was predictably strong. When proton channels were created in the liposome walls, both compounds responded equally to the bulk pH changes. These results suggest that porphyrin dendrimers can be used as fluorescent pH indicators for proton gradient measurements.


Acute Lower Gastrointestinal Bleeding: Temporal Factors Associated With Positive Findings on Catheter Angiography After (99m)Tc-Labeled RBC Scanning.
The objective of the study was to determine if time to positive (TTP), defined as the time from the start of (99m)Tc-labeled RBC scanning to the appearance of a radionuclide blush (considered to be a positive finding for acute lower gastrointestinal bleeding [LGIB]), and lag time (LT), defined as the time from the appearance of a radionuclide blush to the start of catheter angiography (CA), affected the yield of CA for the detection of acute LGIB. TTP and LT were retrospectively evaluated in 120 patients who had positive findings for acute LGIB on (99m)Tc-labeled RBC scanning and subsequently underwent CA for the diagnosis and localization of gastrointestinal bleeding. Two nuclear medicine fellowship-trained radiologists independently reviewed the (99m)Tc-labeled RBC scans. Two fellowship-trained interventional radiologists independently reviewed the angiograms. All data were analyzed using SAS software. When a TTP threshold of ≤ 9 minutes was used, the sensitivity, specificity, positive predictive value, and negative predictive value for a positive CA study were 92%, 35%, 27%, and 94%, respectively. In addition, the odds of detecting bleeding on CA increased 6.1-fold with a TTP of ≤ 9 minutes relative to a TTP of > 9 minutes (p = 0.020). A significant inverse relationship was found between LT and a positive CA study (p = 0.041). TTP and LT impact the rate of positive CA studies. A TTP threshold of ≤ 9 minutes allows the detection of almost all patients who would benefit from CA for treatment and allows a reduction in unnecessary negative CA studies. The likelihood of positive findings on CA decreases with a delay in the performance of CA.


[Association of phosphoinositide-specific phospholipase Cgamma1 with elements of cytoskeleton in A-431 cells].
It is shown that phosphoinositide-specific phospholipase C gamma 1 (PLC gamma 1), a substrate of growth factor receptors, is associated with cytoskeleton in A-431 cells. PLC gamma 1 is co-localized only with the cortical actin but not with actin stress fibers. Since EGF receptor is also co-localized with the cortical actin it is concluded that PLC gamma 1 co-localized with actin is mediated by the EGF receptor. After the treatment with cytochalasin B PLC gamma 1 is co-localized with actin aggregates and cytoskeleton elements other than actin. Using double immunofluorescence PLC gamma 1 is shown to be associated with cytokeratin intermediate filaments. The cross-talking of different cytoskeleton elements and their participation in cell signaling is discussed.


Differentiation of Neisseria gonorrhoeae IB-3 and IB-7 serovars by direct sequencing of protein IB gene and pulsed-field gel electrophoresis.
Serotyping of Neisseria gonorrhoeae based on the differential recognition by a panel of six monoclonal antibodies (MAbs) against Protein IB (PIB) is a valuable tool for studying the epidemiology of gonorrhoea. However, the predominance of certain serovars in specific geographic regions necessitates the development of new MAbs or new genotyping approaches. Nucleotide and amino-acid sequence analysis of PIB from strains within the IB-3 and IB-7 serovars revealed strain differences within the same serovar. Based on the generation of PIB nucleotide and deduced amino-acid sequences that centred on amino-acid residues 196 and 237, eight serovar IB-3 strains and nine serovar IB-7 strains were separately subdivided into five groups. Intra-serovar differences were also established by pulsed-field gel electrophoresis (PFGE) of macro-restriction fragments generated by SpeI- and NheI-cleavage of genomic DNA. There was good correlation between the results based on PIB gene PCR-sequencing and those based on PFGE analysis of macro-restriction fragment patterns. These data demonstrate the high precision of PFGE analysis and indicate that this approach can be used as a rapid epidemiological subtyping system for major serovars of N. gonorrhoeae.


Binding Cooperativity Matters: A GM1-Like Ganglioside-Cholera Toxin B Subunit Binding Study Using a Nanocube-Based Lipid Bilayer Array.
Protein-glycan recognition is often mediated by multivalent binding. These multivalent bindings can be further complicated by cooperative interactions between glycans and individual glycan binding subunits. Here we have demonstrated a nanocube-based lipid bilayer array capable of quantitatively elucidating binding dissociation constants, maximum binding capacity, and binding cooperativity in a high-throughput format. Taking cholera toxin B subunit (CTB) as a model cooperativity system, we studied both GM1 and GM1-like gangliosides binding to CTB. We confirmed the previously observed CTB-GM1 positive cooperativity. Surprisingly, we demonstrated fucosyl-GM1 has approximately 7 times higher CTB binding capacity than GM1. In order to explain this phenomenon, we hypothesized that the reduced binding cooperativity of fucosyl-GM1 caused the increased binding capacity. This was unintuitive, as GM1 exhibited higher binding avidity (16 times lower dissociation constant). We confirmed the hypothesis using a theoretical stepwise binding model of CTB. Moreover, by taking a mixture of fucosyl-GM1 and GM2, we observed the mild binding avidity fucosyl-GM1 activated GM2 receptors enhancing the binding capacity of the lipid bilayer surface. This was unexpected as GM2 receptors have negligible binding avidity in pure GM2 bilayers. These unexpected discoveries demonstrate the importance of binding cooperativity in multivalent binding mechanisms. Thus, quantitative analysis of multivalent protein-glycan interactions in heterogeneous glycan systems is of critical importance. Our user-friendly, robust, and high-throughput nanocube-based lipid bilayer array offers an attractive method for dissecting these complex mechanisms.


Absolute quality evaluation of protein model structures using statistical potentials with respect to the native and reference states.
In protein structure prediction, it is crucial to evaluate the degree of native-likeness of given model structures. Statistical potentials extracted from protein structure data sets are widely used for such quality assessment problems, but they are only applicable for comparing different models of the same protein. Although various other methods, such as machine learning approaches, were developed to predict the absolute similarity of model structures to the native ones, they required a set of decoy structures in addition to the model structures. In this paper, we tried to reformulate the statistical potentials as absolute quality scores, without using the information from decoy structures. For this purpose, we regarded the native state and the reference state, which are necessary components of statistical potentials, as the good and bad standard states, respectively, and first showed that the statistical potentials can be regarded as the state functions, which relate a model structure to the native and reference states. Then, we proposed a standardized measure of protein structure, called native-likeness, by interpolating the score of a model structure between the native and reference state scores defined for each protein. The native-likeness correlated with the similarity to the native structures and discriminated the native structures from the models, with better accuracy than the raw score. Our results show that statistical potentials can quantify the native-like properties of protein structures, if they fully utilize the statistical information obtained from the data set.


The theory of homeoviscous adaptation of membranes applied to deep-sea animals.
Deep-sea organisms live at high pressure and low temperature. These two factors increase the order of the bilayer in cell membranes, and, on the basis of the homeoviscous theory of membrane adaptation, predictions are made of the fluidity and composition of deep-sea membranes. Some of the predictions have been tested using membranes from deep-sea fish and some of the results are consistent with the theory. The conclusion is that the theory receives general support from the deep sea and, conversely, adaptation of membranes to high pressure can be profitably investigated in a more detailed manner by testing further predictions based on the homeoviscous concept.


Colonic perforation following endoscopic retrograde cholangiopancreatography-associated pancreatitis.
We highlight a potentially lethal complication of acute severe pancreatitis that may not be suspected in severely ill patients. A 41-year-old woman developed acute severe pancreatitis following endoscopic retrograde cholangiopancreatography (ERCP) for suspected choledocholithiasis. When her condition deteriorated dramatically after 2 weeks of intensive medical management, a computed tomography scan and water-soluble contrast enema examination revealed a large colonic perforation and associated collection in the right flank. Surgical management included an extended right hemicolectomy and a second laparotomy to attend to soiling at the surgical site. She survived and was discharged. Colonic perforation is a recognised complication of pancreatitis that carries a high mortality. It may result from a combination of ischaemia to the colon and a direct effect of noxious pancreatic enzymes. Almost all cases are only diagnosed at laparotomy. In view of the current trend of non-surgical management of pancreatitis and associated complications, colonic perforation should be considered in patients who deteriorate or fail to improve. To our knowledge this is the first case of a secure pre-operative diagnosis of colonic perforation due to to pancreatitis.


In vivo Efficacy and Pharmacokinetics of Optimized Apidaecin Analogs.
Proline-rich antimicrobial peptides (PrAMPs) represent promising alternative therapeutic options for the treatment of multidrug-resistant bacterial infections. PrAMPs are predominantly active against Gram-negative bacteria by inhibiting protein expression via at least two different modes of action, i.e., blocking the ribosomal exit tunnel of 70S ribosomes (oncocin-type binding) or inhibiting the assembly of the 50S ribosomal subunit (apidaecin-type binding). The in vivo efficacy and favorable biodistribution of oncocins confirmed the therapeutic potential of short PrAMPs for the first time, whereas the in vivo evaluation of apidaecins is still limited despite the promising efficacy of apidaecin-analog Api88 in an intraperitoneal murine infection model. Here, the in vivo efficacy of apidaecin-analog Api137 was studied, which rescued all NMRI mice from a lethal intraperitoneal infection with E. coli ATCC 25922 when administered three times intraperitoneal at doses of 0.6 mg/kg starting 1 h after infection. When Api88 and Api137 were administered intravenous or intraperitoneal at doses of 5 and 20 mg/kg, their plasma levels were similarly low (<3 μg/mL) and four-fold lower than for oncocin-analog Onc72. This contradicted earlier expectation based on the very low serum stability of Api88 with a half-life time of only ~5 min compared to ~6 and ~3 h for Api137 and Onc72, respectively. Pharmacokinetic data relying on a sensitive mass spectrometry method utilizing multiple reaction monitoring and isotope-labeled peptides revealed that Api88 and Api137 were present in blood, urine, and kidney, and liver homogenates at similar levels accompanied by the same major metabolites comprising residues 1-16 and 1-17. The pretended discrepancy was solved, when all peptides were incubated in peritoneal lavage. Api137 was rapidly degraded at the C-terminus, while Api88 was rather stable despite releasing the same degradation products. Onc72 was very stable explaining its higher plasma levels compared to Api88 and Api137 after intraperitoneal administration illuminating its good in vivo efficacy. The data indicate that the degradation of therapeutic peptides should be studied in serum and further body fluids. Moreover, the high efficacy in murine infection models and the fast clearance of Api88 and Api137 within ~60 min after intravenous and ~90 min after intraperitoneal injections indicate that their in vivo efficacy relates to the maximal peptide concentration achieved in blood.


The role of perioperative blood transfusion on postoperative outcomes and overall survival in patients after laparoscopic radical cystectomy.
There are different studies in the last decades focused on general surgery patients and the impact of perioperative blood transfusion (PBT) in cancer patients' survival, and most of them have supported an independent association between PBT and worse survival in those with solid tumor malignancies. The aim of this study is to evaluate the impact of perioperative blood transfusion on the postoperative outcomes and survival of patients after LRC. We performed a retrospective study analyzing our series of 218 patients surgically treated with LRC form 2005 to 2012. One-way analysis of variance test was used. Survival was estimated using the Kaplan - Meier method and was compared with log - rank and the Cox regression model was used to evaluate the association of PBT with the outcomes. The PBT rate of LRC series was 16%. Patients' age, comorbidities and pathological stage were not related to the PBT rate. A statistically significant relationship was found between the PBT rate and the appearance of infectious complications. Overall 3 years survival estimated with the Kaplan-Meier method was significantly worse in the transfused group: 41.38% versus 63.57% for non-transfused patients. PBT was not a significant independent predictor factor in the survival of patients after LRC. The main independent factor was the TNM classification. Many studies including ours have reported a lower survival rate in patients who receive PBT after oncological surgery. There was a relationship between infectious complications and PBT. We have to make efforts to limit the use of blood products in patients surgically treated with radical cystectomy for bladder cancer.


Predictors and prognosis of delirium among older subjects in cardiac intensive care unit: focus on potentially preventable forms.
Delirium is a common and potentially preventable condition in older individuals admitted to acute and intensive care wards, associated with negative prognostic effects. Its clinical relevance is being increasingly recognised also in cardiology settings. The aim of the present study was to assess the prevalence, incidence, predictors and prognostic role of delirium in older individuals admitted to two cardiology intensive care units. All patients aged over 65 years consecutively admitted to the two participating cardiology intensive care units were enrolled. Assessment on admission included acute physiological derangement (modified rapid emergency medicine score, REMS), chronic comorbidity, premorbid disability and dementia. The Confusion Assessment Method-Intensive Care Unit was applied daily for delirium detection. Of 497 patients (40% women, mean age 79 years), 18% had delirium over the entire cardiology intensive care unit course, half of whom more than 24 hours after admission (incident delirium). Advanced age, a main diagnosis of ST-segment elevation myocardial infarction or acute respiratory failure, modified REMS, comorbidity and dementia were independent predictors of delirium. Adjusting for patient's features on admission, incident delirium was predicted by invasive procedures (insertion of peripheral arterial catheter, urinary catheter, central venous catheter, naso-gastric tube and intra-aortic balloon pump). In a logistic regression model, delirium was an independent predictor of inhospital mortality (odds ratio 3.18, 95% confidence interval 1.02, 9.93). Eighteen per cent of older cardiology intensive care unit patients had delirium, with half of the cases being incident, thus potentially preventable. Invasive procedures were independently associated with incident delirium. Delirium was an independent predictor of inhospital mortality. Awareness of delirium should be increased in the cardiology intensive care unit setting and prevention studies are warranted.


Evaluation of preservative effectiveness of liquid crystalline systems with retynil palmitate by the challenge test and D-value.
The objective of this work was to evaluate the preservative effectiveness of liquid crystalline systems containing retynil palmitate (RP) by the challenge test (CT) and D-value. A system was developed containing water, silicon glycol copolymer, and polyether functional siloxane with 1% RP added. The analyses were carried out by methods in the U.S. Pharmacopeia (USP 31, 2008) using the microorganisms Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger. The CT showed that after 7 days, all microorganisms were eliminated except A. niger, which maintained viability for at least 28 days after inoculation. Moreover, the microorganisms E. coli, P. aeruginosa, S. aureus, C. albicans, and A. niger presented different growth behaviors, evidenced by differences among the D-values calculated. It was concluded that the CT and D-value were efficient methods for evaluation of the preservative property of these formulations.


Osteoarthritis year in review 2013: biology.
The purpose of this review was to present highlights from the published literature on the topic of the biology of osteoarthritis (OA). A PubMed search was conducted in order to locate original research manuscripts published since the last OARSI meeting in 2012. From review of the published literature, common themes emerged as active areas of research over the past year including studies in the areas of epigenetics, Wnt signaling, the role of inflammatory pathways in OA, lubricin, fibroblast growth factor signaling, and studies on OA biology in bone. Key findings in these areas were summarized and implications for future therapies were discussed.


The effects of eccentric exercise on muscle function and proprioception of individuals being overweight and underweight.
The aim of this study was to estimate the effect of being overweight or underweight on proprioception at rest and after muscle damaging eccentric exercise. Twelve lean, 12 overweight, and 8 underweight female participants performed an eccentric exercise session using the knee extensor muscles of the dominant leg. Muscle damage indices and proprioception were assessed up to 3 days postexercise. The results indicated that proprioception at baseline of the lean individuals was superior to that of the other 2 groups. The overweight individuals exhibited a smaller knee joint reaction angle to release than did the lean group, whereas the underweight individuals exhibited a larger reaction angle to release than did the lean group. After eccentric exercise, proprioception was affected more in the overweight and the underweight groups than in the lean group. The greater exercise-induced muscle damage appeared in the overweight group, and the deficient muscle mass of the underweight participants could explain in part the greater disturbances that appeared in proprioception in these 2 groups than for the lean counterparts. In conclusion, deviating from the normal body mass is associated with significant disturbances in the proprioception of the legs at rest and after participation in activities involving eccentric actions.


[Ultrastructural characteristic of reactively modified myocardium cultured in vitro].
The aim of the present investigation was to study the structure of reactively modified rat myocardium (experimental heart failure model) during in vitro organotypical culture, including the direct effect of some hypothalamic nonapeptides (oxytocin) and monoamines (dopamine, noradrenalin). Light and electron microscopic methods were used. The major regularities of structural and functional reorganization of myocardium were determined. Multidirectional nature of ultrastructural changes of cardiomyocytes and of cardiac non-muscular cells was found to depend on direct effect of hypothalamic nonapeptides and monoamines upon myocardium. Adaptogenic role of oxytocin in provision of cardiac muscle protection and vitality is discussed.


MBD3L2 interacts with MBD3 and components of the NuRD complex and can oppose MBD2-MeCP1-mediated methylation silencing.
MBD2 and MBD3 are two proteins that contain methyl-CpG binding domains and have a transcriptional repression function. Both proteins are components of a large CpG-methylated DNA binding complex named MeCP1, which consists of the nucleosome remodeling and histone deacetylase complex Mi2-NuRD and MBD2. MBD3L2 (methyl-CpG-binding protein 3-like 2) is a protein with substantial homology to MBD2 and MBD3, but it lacks the methyl-CpG-binding domain. Unlike MBD3L1, which is specifically expressed in haploid male germ cells, MBD3L2 expression is more widespread. MBD3L2 interacts with MBD3 in vitro and in vivo, co-localizes with MBD3 but not MBD2, and does not localize to methyl-CpG-rich regions in the nucleus. In glutathione S-transferase pull-down assays, MBD3L2 is found associated with several known components of the Mi2-NuRD complex, including HDAC1, HDAC2, MTA1, MBD3, p66, RbAp46, and RbAp48. Gel shift experiments with nuclear extracts and a CpG-methylated DNA probe indicate that recombinant MBD3L2 can displace a form of the MeCP1 complex from methylated DNA. MBD3L2 acts as a transcriptional repressor when tethered to a GAL4-DNA binding domain. Repression by GAL4-MBD3L2 is relieved by MBD2 and vice versa, and repression by MBD2 from a methylated promoter is relieved by MBD3L2. The data are consistent with a role of MBD3L2 as a transcriptional modulator that can interchange with MBD2 as an MBD3-interacting component of the NuRD complex. Thus, MBD3L2 has the potential to recruit the MeCP1 complex away from methylated DNA and reactivate transcription.


Experimental models of cardiac transplantation: design determines relevance.
Experimental models have contributed enormously to basic immunology. However, the use of reductionist experiments has produced results that are not always successfully translated into the clinic. Recently, incorporation of more realistic clinical parameters in experimental designs has produced new insights relevant to cardiac transplantation. Experiments in mice have provided crucial insights into the concept that T cell responses to pathogens generate memory cells with cross-reactive specificities for histocompatibility antigens. These memory T cells are resistant to current immunosuppressive strategies. Memory T cells infiltrate grafts within hours after transplantation, and grafts subjected to clinically relevant periods of cold ischemia are more susceptible to injury by this cellular infiltrate. Early immune responses now can be investigated with improved 'humanized' mice. Mice with multiple knock-in genes for human cytokines support development of human monocytes, macrophages and natural killer cells in increased numbers and with better function. Better and more clinically relevant experimental designs are providing animal models tailored to address clinic exigencies.


Physician responses to a community-level trial promoting judicious antibiotic use.
In an environment of multiple campaigns promoting judicious antibiotic use in children, identification of effective strategies is important. We assessed physician responses to a community-level intervention with respect to antibiotic prescribing, related practices, and perceived effectiveness. This study was a mixed qualitative and quantitative evaluation of a randomized controlled community-wide educational intervention in 16 Massachusetts communities. Physicians in intervention communities received locally endorsed guidelines, group educational sessions, and biweekly newsletters. Parents simultaneously received materials in physicians' offices and by mail. After the intervention, we conducted a mailed physician survey and individual interviews to assess the impact of the intervention. We compared survey responses for intervention and control physicians, and we analyzed interview transcripts to provide in-depth information about selected topics. Among survey respondents (n = 168), 91% of intervention and 4% of control physicians reported receiving intervention materials. Physicians received information from multiple other sources. More intervention than control physicians reported decreased antibiotic prescribing from 2000-2003 (75% vs 58%, P = .03), but there were no differences between groups in knowledge, attitudes, or behaviors favoring judicious antibiotic use. Both groups were concerned about antibiotic resistance and reported room to reduce their own prescribing. Interviewed physicians suggested frequent repetition of messages, brief written materials on specific topics for themselves and patients, and promotion in the mass media as the most effective strategies to reduce prescribing. In multiple communities an intervention in physician offices to promote judicious antibiotic prescribing reached its intended audience, but physicians' self-reported attitudes and practices were similar in intervention and control communities. Campaigns that repeat brief, consistent reminders to multiple stakeholder groups may be most effective at assuring judicious antibiotic use.


White grapes arose through the mutation of two similar and adjacent regulatory genes.
Most of the thousands of grapevine cultivars (Vitis vinifera L.) can be divided into two groups, red and white, based on the presence or absence of anthocyanin in the berry skin, which has been found from genetic experiments to be controlled by a single locus. A regulatory gene, VvMYBA1, which could activate anthocyanin biosynthesis in a transient assay, was recently shown not to be transcribed in white berries due to the presence of a retrotransposon in the promoter. We have found that the berry colour locus comprises two very similar genes, VvMYBA1 and VvMYBA2, located on a single bacterial artificial chromosome. Either gene can regulate colour in the grape berry. The white berry allele of VvMYBA2 is inactivated by two non-conservative mutations, one leads to an amino acid substitution and the other to a frame shift resulting in a smaller protein. Transient assays showed that either mutation removed the ability of the regulator to switch on anthocyanin biosynthesis. VvMYBA2 sequence analyses, together with marker information, confirmed that 55 white cultivars all contain the white berry allele, but not red berry alleles. These results suggest that all extant white cultivars of grape vines have a common origin. We conclude that rare mutational events occurring in two adjacent genes were essential for the genesis of the white grapes used to produce the white wines and white table grapes we enjoy today.


Idle peripheral intravenous cannulation: an observational cohort study of pre-hospital and emergency department practices.
Unused ('idle') peripheral intravenous catheters (PIVC) are those not used within 24 hours of insertion. There is little data on cannulation practices and idle PIVC rates in emergency settings, especially the pre-hospital environment. This was an observational cohort study set in south-east Queensland, Australia using data from a large tertiary level emergency department (ED) and the local statutory ambulance service. Demographic, clinical and PIVC data were collected over two periods; 9 February-18 March 2017 and 5 January-4 February 2018. Adult patients were included if they were allocated an Australasian triage scale (ATS) category between 2 and 5, and had a PIVC inserted in the pre-hospital setting or ED. PIVC use was defined as idle if no fluids, medications or contrast were administered intravenously within 24 hours of insertion. Comparisons between pre-hospital and ED practice and idle PIVC status were undertaken using descriptive statistics and logistic regression. A total of 1249 patients with a PIVC (372 pre-hospital; 877 ED) were included. Overall, 366 PIVCs (29.3%; 95% CI 26.9%-31.9%) remained idle at 24 hours. In the pre-hospital group, 147 (39.5%) PIVCs inserted were not used pre-hospital, and 74 (19.9%) remained idle. In comparison, 292 (33.3%) PIVCs placed in the ED remained idle. ED staff more frequently inserted PIVCs in the antecubital fossa than paramedics (65.5% vs. 49.7%), where forearm PIVC insertion was more common pre-hospital than in ED (13.7% vs. 7.4%). Nursing staff inserted idle PIVCs at a rate of (35.1%) compared to doctors (29.6%) and paramedics (19.9%). Having a PIVC inserted in the ED was the only factor significantly (p ≤ .001) predicting an idle outcome (Odds Ratio: 2.4; 95% CI 1.7-3.3). One-third of PIVCs inserted within the emergency setting remained idle, suggesting unnecessary risk and costs. Pre-hospital and ED PIVC insertion practices differed, with idle PIVCs 2.4 times more prevalent if inserted in the ED than pre-hospital and with greater use of antecubital insertion. Reasons for these differences are not well understood and requires more targeted research.


The lived experience of art making as a companion to the mental health recovery process.
Art making is a common activity provided for consumers in mental health psychosocial rehabilitation services, yet there is little evidence available which examines its role in the recovery process. The current study inquires into mental health consumers' lived experiences of art making within psychosocial rehabilitation services and their views on how art making supports mental health recovery. This research used qualitative in-depth interviews to explore the role of art making in the mental health recovery journey. The sample comprised 18 consumer participants who attended art-based programs in two psychosocial services in Victoria, Australia. The 60-90 min interviews were analysed using interpretative phenomenological analysis. A total of 11 major themes were identified and organised into three areas: qualities conducive to the art making context, how the art making process benefits mental health recovery, and how the image or art product benefits mental health recovery. The 11 themes are described and illustrated from participant interviews. Consumers described art making as a transformative activity which enabled them to take greater control of their lives, resulting in feeling stronger, more confident, and more capable of driving their journey of recovery. The art product also served valuable roles in supporting their recovery. Art making is a highly valued activity by consumers, who suggest that innovative and strengths-based methods, such as art making, can facilitate recovery and self-expression. A key challenge for the field is to determine how such methods can be better integrated into mental health service delivery.


Pretreatment with intravenous ascorbic acid preserves endothelial function during acute hyperglycaemia (R1).
1. Acute hyperglycaemia may impair endothelial function. Ascorbic acid (AA), administered intra-arterially, has been reported to improve endothelium-dependent vasodilatation during a forearm hyperglycaemic clamp. Using a randomized, double-blind, placebo-controlled, cross-over study, we investigated the potential for intravenous ascorbic acid to modify the endothelial response to acute systemic hyperglycaemia in humans. 2. Nine healthy male volunteers were recruited from the hospital staff. Endothelial function was determined by measuring the forearm blood flow responses to intrabrachial infusions of endothelium-dependent (ED) and endothelium-independent (EID) vasodilators. The endothelial function index (EFI) was derived from the ratio of ED and EID vasodilatation. Haemodynamic and endothelial function measurements were performed at baseline and then repeated 2 h after a systemic hyperglycaemic clamp (14 mmol/L). The subjects, studied on two separate occasions, were randomized to placebo or 2 g intravenous ascorbic acid prior to the initiation of hyperglycaemia. 3. After systemic hyperglycaemia with placebo pretreatment, the EFI fell from 1.08 +/- 0.21 to 0.74 +/- 0.13 (difference (95% confidence interval): 0.34 (0.20, 0.47); P < 0.001). When subjects were pretreated with ascorbic acid, the EFI was not affected by hyperglycaemia (1.11 +/- 0.21 to 1.12 +/- 0.17; P = 0.938). This difference between placebo and ascorbic acid was significant (P < 0.001). Plasma ascorbate concentrations decreased during hyperglycaemia and correlated directly with the reduction in the EFI (r = 0.798; P < 0.001). 4. Pretreatment with an intravenous bolus of ascorbic acid can prevent endothelial dysfunction during acute systemic hyperglycaemia. Therefore, ascorbic acid may have potential therapeutic use in clinical situations where acute hyperglycaemia may be a complication.


Would the Siblings of Latinx Men Who Have Sex With Men Encourage Their Brothers to Use PrEP? Findings From a Feasibility and Acceptability Study.
We explored whether siblings can be engaged in PrEP promotion. We used the Information-Motivation-Behavior model to develop and conduct surveys and dyadic interviews with Latino men who have sex with men (LMSM) and their siblings (n = 31) and three sibling-only focus groups (n = 20). For LMSM, only n = 14 (45%) agreed they would benefit from taking PrEP, yet n = 22 (71%) would take PrEP to make their sibling worry less about them, and n = 23 (74%) requested a PrEP referral. For siblings, n = 20 (65%) believed PrEP would benefit their brother, and n = 19 (95%) in the focus groups said they would take PrEP to help their brother get started. Qualitative results include (1) siblings' support for PrEP use, (2) explicit conversations about sex were not necessary for discussing sexual health, and (3) siblings wanted to understand what they could do to encourage their brother to consider PrEP. We conclude siblings can be engaged in PrEP promotion.


A child with ictal vocalizations and generalized epilepsy.
Ictal vocalizations in the form of both articulate speech and non-speech vocalizations have been described in focal epilepsies, with seizures originating mainly from the frontal and temporal lobe, however, this phenomenon has not been described in generalized epilepsies. We report the case of an adolescent boy with juvenile-onset generalized epilepsy who presented with ictal "ovine vocalizations" (resembling the bleating of sheep). The ictal EEG revealed a clear correlate of vocalizations with time-locked generalized spikes and polyspike discharges. The 3T cerebral MRI ruled out any focal lesion. The boy is currently seizure-free under valproic acid, after twelve months of follow-up. We conclude that ictal non-speech vocalizations may be observed not only in focal or structural epilepsies, but also in generalized epilepsies; the exact underlying mechanism of this phenomenon needs to be further delineated. [Published with video sequence].


The Medical Action Ontology: A Tool for Annotating and Analyzing Treatments and Clinical Management of Human Disease.
Navigating the vast landscape of clinical literature to find optimal treatments and management strategies can be a challenging task, especially for rare diseases. To address this task, we introduce the Medical Action Ontology (MAxO), the first ontology specifically designed to organize medical procedures, therapies, and interventions in a structured way. Currently, MAxO contains 1757 medical action terms added through a combination of manual and semi-automated processes. MAxO was developed with logical structures that make it compatible with several other ontologies within the Open Biological and Biomedical Ontologies (OBO) Foundry. These cover a wide range of biomedical domains, from human anatomy and investigations to the chemical and protein entities involved in biological processes. We have created a database of over 16000 annotations that describe diagnostic modalities for specific phenotypic abnormalities as defined by the Human Phenotype Ontology (HPO). Additionally, 413 annotations are provided for medical actions for 189 rare diseases. We have developed a web application called POET (https://poet.jax.org/) for the community to use to contribute MAxO annotations. MAxO provides a computational representation of treatments and other actions taken for the clinical management of patients. The development of MAxO is closely coupled to the Mondo Disease Ontology (Mondo) and the Human Phenotype Ontology (HPO) and expands the scope of our computational modeling of diseases and phenotypic features to include diagnostics and therapeutic actions. MAxO is available under the open-source CC-BY 4.0 license (https://github.com/monarch-initiative/MAxO).


Developmental genetics in ophthalmology.
Much of our knowledge about the function of genes in mammalian development has been derived from the molecular analysis of spontaneous or induced mutations in the mouse. Since mutations affecting the mouse eye can be easily identified, a remarkable number of mutant lines provide animal models for congenital anomalies in man. To understand the mechanisms of lens development in detail, the isolation of the corresponding genes and the characterization of the mutations at the molecular level are important. A prerequisite for molecular analysis is the chromosomal localization of the gene. In this review, some mutants from our institute will be discussed according to the embryological time scale of the expression of the affected genes, reflecting also their genetic hierarchy. (1) In the aphakia mouse mutant, two deletions in the promoter of the homeobox transcription factor Pitx3 lead to a loss of its function and to an arrest of eye development at the lens stalk stage. Mutations in the homologous human PITX3 gene have been demonstrated to be causative of cataracts and the dysmorphology of the anterior segment of the eye. (2) Connexin50 is present in the lens vesicle. Later on, it becomes abundant in the anterior part of the fiber cells and in the lens epithelial cells. Mutations in the connexin50-encoding gene Gja8 lead to dominant cataracts. (3) alphaA-crystallin is present in the mouse lens cup, in the posterior half of the lens vesicle, and later in a high concentration in the lens fiber cells. Mutations in the alphaA-crystallin-encoding gene Cryaa lead to recessive and dominant cataracts. (4) Mutations in the gamma-crystallin -encoding genes (Cryg) are the most frequent cause of congenital, dominant nuclear, or total cataracts in the mouse. Indications from our first studies in congenital human cataracts support these data. (5) Some postnatal, progressive cataracts have been characterized by mutations in the beta-crystallin -encoding genes (Cryb). Since at least one of them is also expressed in the retina and the brain, effects on these tissues have to be considered, too.


Mixed Diffuse and Tumoral Form of Bing-Neel Syndrome Successfully Treated with Ibrutinib.
Bing-Neel syndrome (BNS) is a rare and heterogenous manifestation of Waldenström macroglobulinemia (WM) involving central nervous system (CNS) infiltration by malignant lymphoplasmacytic cells. Efforts to standardize diagnostic criteria have improved in recent years, as have treatment options including the use of the Bruton tyrosine kinase inhibitor (BTKI) ibrutinib. Here, we present the case of a 70-year-old male with a remote history of WM previously treated with bendamustine and rituximab, who presented to medical attention with several months of left-sided weakness, headache, and ataxia. Brain magnetic resonance imaging revealed numerous enhancing masses in the bilateral cerebral hemispheres, inferior medulla, and upper cervical spine. Laboratory studies showed serum IgM lambda monoclonal gammopathy and elevated free serum kappa and lambda light chains, while cerebrospinal fluid flow cytometry revealed CD19+ B cells. Stereotactic brain biopsy of a right frontal brain lesion was consistent with lymphoplasmacytic lymphoma, confirmed by a positive MYD88 L265P mutation. He received ibrutinib 420 mg orally daily, and this resulted in appreciable clinical and radiologic responses, which have persisted over a 31-month period. The advent of molecularly targeted agents and novel therapies for WM has provided patients and clinicians with additional therapeutic options. The use of BTK inhibitors with their high-level CNS penetrance, in particular, offers a novel way to treat BNS and improve patient overall survival while maintaining a high level of quality of life. We discuss the importance of MYD88 L265P testing in the context of BNS as well as the expanding role of BTKIs in treating this disease.


Comparative SEM study on the effect of different demineralization methods with tetracycline HCl on healthy root surfaces.
Periodontal regeneration through the use of root demineralization received a lot of interest in periodontology. Topical application of acid to dentin surfaces produced a zone of demineralization, exposing dentin collagen fibrils and opening dentin tubules. In this study, the in vitro effects of different tetracycline HCl application techniques were investigated. According to the results of this SEM study, it may be desirable to apply tetracycline HCl using burnishing technique to expose maximum intertubular fibrils and for the tubular openings. However, this technique should be studied when placed in an in vivo system.


Deep Learning-Based Quantitative Blastocyst Assessment.
Selecting the single best blastocyst based on morphological appearance for implantation is a crucial part of in vitro fertilization (IVF). Various deep learning and computer vision-based methods have recently been applied for assessing blastocyst quality. However, to the best of our knowledge, most previous works utilize classification networks to give a qualitative evaluation. It would be challenging to rank blastocyst quality with the same qualitative result. Thus, this paper proposes a regression network combined with a soft attention mechanism for quantitatively evaluating blastocyst quality. The network outputs a continuous score to represent blastocyst quality precisely rather than some categories. As to the soft attention mechanism, the attention module in the network outputs an activation map (attention map) localizing the regions of interest (ROI, i.e., inner cell mass (ICM)) of microscopic blastocyst images. The generated activation map guides the entire network to predict ICM quality more accurately. The experimental results demonstrate that the proposed method is superior to traditional classification-based networks. Moreover, the visualized activation map makes the proposed network decision more reliable.


Dynamic localization of a helper NLR at the plant-pathogen interface underpins pathogen recognition.
Plants employ sensor-helper pairs of NLR immune receptors to recognize pathogen effectors and activate immune responses. Yet, the subcellular localization of NLRs pre- and postactivation during pathogen infection remains poorly understood. Here, we show that NRC4, from the "NRC" solanaceous helper NLR family, undergoes dynamic changes in subcellular localization by shuttling to and from the plant-pathogen haustorium interface established during infection by the Irish potato famine pathogen Phytophthora infestans. Specifically, prior to activation, NRC4 accumulates at the extrahaustorial membrane (EHM), presumably to mediate response to perihaustorial effectors that are recognized by NRC4-dependent sensor NLRs. However, not all NLRs accumulate at the EHM, as the closely related helper NRC2 and the distantly related ZAR1 did not accumulate at the EHM. NRC4 required an intact N-terminal coiled-coil domain to accumulate at the EHM, whereas the functionally conserved MADA motif implicated in cell death activation and membrane insertion was dispensable for this process. Strikingly, a constitutively autoactive NRC4 mutant did not accumulate at the EHM and showed punctate distribution that mainly associated with the plasma membrane, suggesting that postactivation, NRC4 may undergo a conformation switch to form clusters that do not preferentially associate with the EHM. When NRC4 is activated by a sensor NLR during infection, however, NRC4 forms puncta mainly at the EHM and, to a lesser extent, at the plasma membrane. We conclude that following activation at the EHM, NRC4 may spread to other cellular membranes from its primary site of activation to trigger immune responses.