Pericardial Fat and the Risk of Heart Failure.
Obesity is a well-established risk factor for heart failure (HF). However, implications of pericardial fat on incident HF is unclear. This study sought to examine the association between pericardial fat volume (PFV) and newly diagnosed HF. This study ascertained PFV using cardiac computed tomography in 6,785 participants (3,584 women and 3,201 men) without pre-existing cardiovascular disease from the MESA (Multi-Ethnic Study of Atherosclerosis). Cox proportional hazards regression was used to evaluate PFV as continuous and dichotomous variable, maximizing the J-statistic: (Sensitivity + Specificity - 1). In 90,686 person-years (median: 15.7 years; interquartile range: 11.7 to 16.5 years), 385 participants (5.7%; 164 women and 221 men) developed newly diagnosed HF. PFV was lower in women than in men (69 ± 33 cm3 vs. 92 ± 47 cm3; p < 0.001). In multivariable analyses, every 1-SD (42 cm3) increase in PFV was associated with a higher risk of HF in women (hazard ratio [HR]: 1.44; 95% confidence interval [CI]: 1.21 to 1.71; p < 0.001) than in men (HR: 1.13; 95% CI: 1.01 to 1.27; p = 0.03) (interaction p = 0.01). High PFV (≥70 cm3 in women; ≥120 cm3 in men) conferred a 2-fold greater risk of HF in women (HR: 2.06; 95% CI: 1.48 to 2.87; p < 0.001) and a 53% higher risk in men (HR: 1.53; 95% CI: 1.13 to 2.07; p = 0.006). In sex-stratified analyses, greater risk of HF remained robust with additional adjustment for anthropometric indicators of obesity (p ≤ 0.008), abdominal subcutaneous or visceral fat (p ≤ 0.03) or biomarkers of inflammation and hemodynamic stress (p < 0.001) and was similar among Whites, Blacks, Hispanics, and Chinese (interaction p = 0.24). Elevated PFV predominantly augmented the risk of HF with preserved ejection fraction (p < 0.001) rather than reduced ejection fraction (p = 0.31). In this large, community-based, ethnically diverse, prospective cohort study, pericardial fat was associated with an increased risk of HF, particularly HF with preserved ejection fraction, in women and men.


Sustainability assessment on paddy-upland crop rotations by carbon, nitrogen and water footprint integrated analysis: A field scale investigation.
Nutrients of carbon, nitrogen and water of farmland ecosystem are essential foundation to guarantee crop production, but also environmental flows associated greenhouse gas (GHG), reactive nitrogen (Nr) releases, and water consumption. Their flow characteristics serve as a crucial starting point for creating efficient management practices and mitigation measures. Therefore, the objectives of this study are to quantify the carbon footprint (CF), nitrogen footprint (NF), water footprint (WF), and comprehensive environmental footprint (ComF) of six paddy-upland rotation systems, including fallow-paddy rice (FA-PR), Chinese milk vetch-paddy rice (CMV-PR), wheat-paddy rice (WH-PR), rapeseed-paddy rice (RA-PR), green forage wheat-paddy rice (WF-PR), and vicia faba bean-paddy rice (FB-PR), as well as to analysis their relationships and define driving factors. Results showed that the lowest area-scaled CF of 3.74 t CO2-eq ha-1 were observed in the CMV-PR rotation, which were 41% lower than that for WH-PR (the highest CF, 9.13 t CO2-eq ha-1) when soil carbon change was taken into account. It is of importance that soil carbon sequestration in CMV-PR rotation could offset up to around 57% of its CF, while the WH-PR rotation only offset 25%. The RA-PR rotation had the highest area-scaled NF and WF, which was 1.8 and 1.9 times greater than those of the lowest rotation in FA-PR. In terms of comprehensive environmental effects, the six rotation systems showed the order of FA-PR < CMV-PR < FB-PR < RA-PR < WF-PR < WH-PR, with NH3 volatilization accounting 60.7%-66.7% and blue-green WF for 17.5%-26.6% of the total. Therefore, priority should be given to optimizing N fertilizer application and water consumption for paddy-upland rotation systems. The study also suggested that appropriate inter-annual adjustment of rotation system could contribute to achieving GHG mitigations and Nr losses.


Number of children and the risk of obesity in older women.
The purpose of this study is to examine number of live births, other reproductive factors, and health behaviors in relation to obesity risk among older women. Data were collected during in-person and telephone-based interviews from a population-based cohort in Utah. A total of 2,035 women aged 66-102 are included in this report. Overall, 403 (20%) older women were determined to be obese. The rates of obesity were significantly higher with increasing numbers of children, demonstrating a dose-response relationship (P < 0.05). After adjustment for age, education, marital status, BMI at age 18, use of oral contraceptives, hysterectomy status, physical activity, current use of hormone therapy, and age at menarche, the risk of obesity increased 11% with each additional live birth. In additional analyses that excluded nulliparous women, after adjusting for cumulative months of breast-feeding, the risk of obesity increased 7% with each live birth. In this cohort of older women, we found higher rates of obesity with increasing number of children that was independent of socioeconomic status and other confounding factors. In a cohort of older women, higher rates of obesity were associated with increasing number of children that was independent of socioeconomic status and other confounding factors.


[Arterial embolization in the care of massive hemorrhage from the rectum].
Haemorrhage from distal parts of the GIT is as a rule associated with diagnostic and therapeutic difficulties, in particular when massive haemorrhage is involved. The authors describe a group of three patients where the source of haemorrhage was an inoperable rectal tumour. This problematic condition was successfully treated by embolization of the upper rectal artery.


Cardiohepatic Syndrome Is Associated With Poor Prognosis in Patients Undergoing Tricuspid Transcatheter Edge-to-Edge Valve Repair.
The aim of this study was to evaluate the prevalence and prognostic implications of cardiohepatic syndrome (CHS) in patients with tricuspid regurgitation (TR) treated with tricuspid transcatheter edge-to-edge valve repair (T-TEER). The role of CHS in patients undergoing T-TEER for severe TR has not been studied. This study included patients who underwent T-TEER for TR between 2016 and 2020 at 2 high-volume academic centers. CHS was defined as elevation of at least 2 of 3 cholestatic hepatic enzymes. The impact of CHS on 1-year all-cause mortality and clinical outcomes after T-TEER was investigated. T-TEER reduced TR severity to ≤2+ in 257 of 305 included patients (86.2%). CHS was present in 45.2% of patients and was associated with a higher rate of mortality and of first hospitalization for heart failure (HHF) (CHS vs no CHS: estimated 1-year mortality, 34.0% vs 15.9% [P < 0.01]; HHF, 23.0% vs 12.2% [P = 0.01]). CHS was identified as an independent predictor of 1-year all-cause mortality (HR: 1.86; 95% CI: 1.10-3.14; P < 0.05). Irrespective of CHS, T-TEER improved New York Heart Association functional class and 6-minute walk distance in the majority of patients. In patients with impaired baseline hepatic function, laboratory liver parameters improved after T-TEER. CHS is a strong predictor of mortality and HHF after T-TEER and should be evaluated in the process of procedural decision making for T-TEER. Nevertheless, T-TEER is associated with relevant symptomatic alleviation irrespective of CHS.


The utilisation of a structured debriefing framework within the pre-hospital environment: a service evaluation.
Debriefing improves care and reduces error. To be effective, debriefs should be facilitated by trained individuals utilising structured and validated tools. Currently, in UK ambulance services there is no published evidence that structured processes utilising validated tools are being consistently delivered by trained facilitators, potentially impacting clinical practice. A pre-intervention survey related to debriefing was sent to 1000 clinicians within a specific geographical area of the trust via e-mail. In addition, 12 senior or advanced paramedics were recruited from the same area to participate in a training day and 12-week trial, utilising the Debrief Diamond as part of post-event debriefing. Following the trial period, all facilitators and participants of any recorded debriefs were invited to complete a post-intervention survey. A total of 130 staff responded to the pre-intervention survey, with 22% reporting that previous debriefs had not identified areas for learning, and 13% reporting that previous debriefs had not identified good practice, learning opportunities or near misses. Post-intervention, 89% believed the process of debriefing was improved utilising a structured framework, 85% stated trained individuals improved the process, 93% reported the identification of good practice, 70% identified team level learning and 100% of facilitators reported improvements in identifying and supporting learning. Improvements in identifying good practice and learning opportunities were reported by both clinicians and facilitators in this evaluation, reflecting current evidence that structured and facilitated debriefs support safer care through the identification and subsequent reduction of human error. Consequently, the evaluation of appropriate debrief frameworks to provide consistency and validity to clinical debriefs in the pre-hospital environment should be considered to support safer clinical care.


Hydrothermal epitaxy and resultant properties of EuTiO3 films on SrTiO3(001) substrate.
We report a novel epitaxial growth of EuTiO3 films on SrTiO3(001) substrate by hydrothermal method. The morphological, structural, chemical, and magnetic properties of these epitaxial EuTiO3 films were examined by scanning electron microscopy, transmission electron microscopy, high-resolution X-ray diffractometry, X-ray photoelectron spectroscopy, and superconducting quantum interference device magnetometry, respectively. As-grown EuTiO3 films with a perovskite structure were found to show an out-of-plane lattice shrinkage and room-temperature ferromagnetism, possibly resulting from an existence of Eu(3+). Postannealing at 1,000°C could reduce the amount of Eu(3+), relax the out-of-plane lattice shrinkage, and impact the magnetic properties of the films. 81.10.Aj; 81.15.-z; 61.05.-a.


Vestibular function of patients with profound deafness related to GJB2 mutation.
GJB2 mutations are responsible not only for deafness but also for the occurrence of vestibular dysfunction. However, vestibular dysfunction tends to be unilateral and less severe in comparison with that of bilateral deafness. The correlation between the cochlear and vestibular end-organs suggests that some children with congenital deafness may have vestibular impairments. On the other hand, GJB2 gene mutations are the most common cause of nonsyndromic deafness. The vestibular function of patients with congenital deafness (CD), which is related to GJB2 gene mutation, remains to be elucidated. The purpose of this study was to analyze the relationship between GJB2 gene mutation and vestibular dysfunction in adults with CD. A total of 31 subjects, including 10 healthy volunteers and 21 patients with CD, were enrolled in the study. A hearing test and genetic analysis were performed. The vestibular evoked myogenic potentials (VEMPs) were measured and a caloric test was performed to assess the vestibular function. The percentage of vestibular dysfunction was then statistically analyzed. The hearing level of all CD patients demonstrated a severe to profound impairment. In seven CD patients, their hearing impairment was related to GJB2 mutation. Five of the seven patients with CD related to GJB2 mutation demonstrated abnormalities in one or both of the two tests. The percentage of vestibular dysfunction of the patients with CD related to GJB2 mutation was statistically higher than in patients with CD unrelated to GJB2 mutation and in healthy controls.


Survival of patients with chronic heart failure in the community: a systematic review and meta-analysis.
To provide reliable survival estimates for people with chronic heart failure and explain variation in survival by key factors including age at diagnosis, left ventricular ejection fraction, decade of diagnosis, and study setting. We searched in relevant databases from inception to August 2018 for non-interventional studies reporting survival rates for patients with chronic or stable heart failure in any ambulatory setting. Across the 60 included studies, there was survival data for 1.5 million people with heart failure. In our random effects meta-analyses the pooled survival rates at 1 month, 1, 2, 5 and 10 years were 95.7% (95% confidence interval 94.3-96.9), 86.5% (85.4-87.6), 72.6% (67.0-76.6), 56.7% (54.0-59.4) and 34.9% (24.0-46.8), respectively. The 5-year survival rates improved between 1970-1979 and 2000-2009 across healthcare settings, from 29.1% (25.5-32.7) to 59.7% (54.7-64.6). Increasing age at diagnosis was significantly associated with a reduced survival time. Mortality was lowest in studies conducted in secondary care, where there were higher reported prescribing rates of key heart failure medications. There was significant heterogeneity among the included studies in terms of heart failure diagnostic criteria, participant co-morbidities, and treatment rates. These results can inform health policy and individual patient advanced care planning. Mortality associated with chronic heart failure remains high despite steady improvements in survival. There remains significant scope to improve prognosis through greater implementation of evidence-based treatments. Further research exploring the barriers and facilitators to treatment is recommended.


Experimental infection of cattle, sheep, and goats with the newly emerged epizootic hemorrhagic disease virus serotype 8.
Epizootic hemorrhagic disease virus serotype 8 (EHDV-8) emerged in Europe for the first time in late 2022. In this study, we investigated the kinetics of EHDV-8 infection in cattle, sheep, and goats. Following experimental infection with EHDV-8, four out of five calves displayed fever, while another calf exhibited ulcerative and crusty lesions of the muzzle. RNAemia peaked at day 7 post infection in all calves and remained relatively stable till the end of the study, at 78 days post infection. Infectious virus was isolated up to 21 days post infection in one calf. As far as small ruminants are concerned, one sheep experienced fever and two out of five had consistent RNAemia that lasted until the end of the study. Remarkably, infectious virus was evidenced at day 7 post infection in one sheep. In goats, no RNA was observed. All infected animals seroconverted, and a neutralizing immune response was observed in all species, with calves exhibiting a more robust response than sheep and goats. Our study provides insights into the kinetics of EHDV-8 infection and the host immune responses. We also highlight that sheep may also play a role in EHDV-8 epidemiology. Altogether, the data gathered in this study could have important implications for disease control and prevention strategies, providing crucial information to policy makers to mitigate the impact of this viral disease on livestock.


Correction: Gualdani, R.; et al. Store-Operated Calcium Entry Contributes to Cisplatin-Induced Cell Death in Non-Small Cell Lung Carcinoma. Cancers 2019, 11, 430.
The authors wish to make the following corrections to this paper [...].


A study to assess the quality of information in referral letters to the orthodontic department at Kingston Hospital, Surrey.
To assess the quality of information included in referral letters sent to the orthodontic department at Kingston Hospital, Surrey, UK. Referral letters sent by both general dental practitioners (GDPs) and specialist orthodontists were analysed retrospectively in order to ascertain the percentage meeting the inclusion criteria as suggested by Mossey (2006) and the British Orthodontic Society (2008) for the quality of information included in an ideal orthodontic referral letter. Thirty-five consecutive letters sent between May and September 2005 and 206 letters sent in the same period in 2008 were collected by hand and analysed against the inclusion criteria. The numbers of referral letters received from GDPs, specialist orthodontists, and others sources were also determined. Most of the referrals sent in 2005 and 2008 included 40-50% of the referral inclusion points. This was despite an almost twofold increase in the number of referral letters received from specialist orthodontic practitioners in 2008. The majority of the letters, from both GDPs and specialists, did not include details of the oral hygiene or caries status of the patient, or an indication of their motivation towards treatment. None of the referral letters included a plaque score. The main weaknesses in the quality of information provided in referral letters were that in more than 80% of the letters there was no mention of the patient's medical history and no comment on caries status, the standard of oral hygiene, patient motivation for treatment, or an Index of Orthodontic Treatment Need score. The quality of information included in referral letters sent to Kingston Hospital orthodontic department needs to be improved.


Estimating life expectancy adjusted by self-rated health status in the United States: national health interview survey linked to the mortality.
Life expectancy is increasingly incorporated in evidence-based screening and treatment guidelines to facilitate patient-centered clinical decision-making. However, life expectancy estimates from standard life tables do not account for health status, an important prognostic factor for premature death. This study aims to address this research gap and develop life tables incorporating the health status of adults in the United States. Data from the National Health Interview Survey (1986-2004) linked to mortality follow-up through to 2006 (age ≥ 40, n = 729,531) were used to develop life tables. The impact of self-rated health (excellent, very good, good, fair, poor) on survival was quantified in 5-year age groups, incorporating complex survey design and weights. Life expectancies were estimated by extrapolating the modeled survival probabilities. Life expectancies incorporating health status differed substantially from standard US life tables and by health status. Poor self-rated health more significantly affected the survival of younger compared to older individuals, resulting in substantial decreases in life expectancy. At age 40 years, hazards of dying for white men who reported poor vs. excellent health was 8.5 (95% CI: 7.0,10.3) times greater, resulting in a 23-year difference in life expectancy (poor vs. excellent: 22 vs. 45), while at age 80 years, the hazards ratio was 2.4 (95% CI: 2.1, 2.8) and life expectancy difference was 5 years (5 vs. 10). Relative to the US general population, life expectancies of adults (age < 65) with poor health were approximately 5-15 years shorter. Considerable shortage in life expectancy due to poor self-rated health existed. The life table developed can be helpful by including a patient perspective on their health and be used in conjunction with other predictive models in clinical decision making, particularly for younger adults in poor health, for whom life tables including comorbid conditions are limited.


Discrepant results obtained with two versions of the Farnsworth Panel D-15 test in patients with acquired blue-yellow defects.
Two commercially available Panel D-15 tests were administered to two patients with acquired blue-yellow defects. The results obtained with each test were different and apparently influenced by the cap construction which is the only difference between the two tests. In one test the surface of the cap is concave and polished while in the other test the cap surface is flat and has a matte or dull finish. The mechanism accounting for the different results is not obvious.


Is there an ideal way to initiate antiplatelet therapy with aspirin? A crossover study on healthy volunteers evaluating different dosing schemes with whole blood aggregometry.
Guidelines recommend an early initiation of aspirin treatment in patients with acute cerebral ischemia. Comparative studies on the best starting dose for initiating aspirin therapy to achieve a rapid antiplatelet effect do not exist. This study evaluated the platelet inhibitory effect in healthy volunteers by using three different aspirin loading doses to gain a model for initiating antiplatelet treatment in acute strokes patients. Using whole blood aggregometry, this study with a prospective, uncontrolled, open, crossover design examined 12 healthy volunteers treated with three different aspirin loading doses: intravenous 500 mg aspirin, oral 500 mg aspirin, and a course of 200 mg aspirin on two subsequent days followed by a five-day course of 100 mg aspirin. Aspirin low response was defined as change of impedance exceeding 0 Ω after stimulation with arachidonic acid. Sufficient antiplatelet effectiveness was gained within 30 seconds when intravenous 500 mg aspirin was used. The mean time until antiplatelet effect was 74 minutes for 500 mg aspirin taken orally and 662 minutes (11.2 hours) for the dose scheme with 200 mg aspirin with a high inter- and intraindividual variability in those two regimes. Platelet aggregation returned to the baseline range during the wash-out phase within 4 days. Our study reveals that the antiplatelet effect differs significantly between the three different aspirin starting dosages with a high inter- and intraindividual variability of antiplatelet response in our healthy volunteers. To ensure an early platelet inhibitory effect in acute stroke patients, it could be advantageous to initiate the therapy with an intravenous loading dose of 500 mg aspirin. However, clinical outcome studies must still define the best way to initiate antiplatelet treatment with aspirin.


Drying efficiency and particle movement in coating--impact on particle agglomeration and yield.
The purpose of this study was to determine the influences of drying efficiency and particle movement on the degree of agglomeration and yield of pellets coated under different conditions. Thermodynamic conditions were varied using different inlet air temperatures and airflow rates, fluid dynamics were varied using different airflow patterns and air velocities, and two sizes of pellets were coated at different airflow rates and partition gaps. Agglomeration was minimized when all the moisture introduced into the system was removed by the drying air. Excessively dry conditions led to increased loss of yield due to spray-drying effect and attrition. Fluid dynamics were still important even with adequate drying, as the degree of agglomeration was relatively higher in the non-swirling airflow of Wurster coating than in the swirling airflow of precision coating. Increasing air velocities increased pellet velocities, resulting in lower degrees of agglomeration. Hence, agglomeration due to fluid dynamics was attributed to differences in pellet velocities, pellet proximity and pellet trajectories within the partition column. Smaller pellets agglomerated primarily from inadequate drying and not due to inadequate opportunities for particle movement. Larger pellets were more affected by the partition gap due to restriction of their movement through the partition gap. Hence, both thermodynamics and fluid dynamics were found to be important in minimizing agglomeration and ensuring quality coated products.


The involvements of calcium-dependent protein kinases and catechins in tea plant [Camellia sinensis (L.) O. Kuntze] cold responses.
Temperature is one of the most important environmental factors limiting tea plant growth and tea production. Previously we reported that both Ca2+ and ROS signals play important roles in tea plant cold acclimation. Here, we identified 26 CsCPK transcripts, analyzed their phylogenetic and sequence characters, and detected their transcriptions to monitor Ca2+ signaling status. Tissue-specific expression profiles indicated that most CsCPK genes were constitutively expressed in tested tissues, suggesting their possible roles in development. Cold along with calcium inhibitor assays suggested that CsCPKs are important cold regulators and CsCPK30/5/4/9 maybe the key members. Moreover, LaCl3 or EGTA pre-treatment could result in impaired Ca2+ signaling and compromised cold-responding network, but higher catechins accumulation revealed their potential positive roles in cold responses. Those findings indicated that catechins and other secondary metabolites in tea plant may form an alternative cold-responding network that closely correlated with Ca2+ signaling status.


Catecholamines oxidation by xanthine oxidase.
Dopamine and structurally related catecholamines in the presence of hydrogen peroxide are oxidized in vitro by xanthine oxidase producing the corresponding melanin pigments. The kinetic parameters of the reaction, measured as aminochrome formation, have been calculated. The rate of peroxidation depends on enzyme and hydrogen peroxide concentration. The optimum pH for the peroxidative activity of the enzyme is around 8.5. Activation of the peroxidative reaction is also elicited by catechol compounds through a redox cycle mechanism. Implications about the possible biochemical relevance of xanthine oxidase activity on catecholamines oxidation are discussed.


Clinical analysis of 71 spontaneous pneumomediastinum cases: an observational study from a tertiary care hospital in Japan.
Spontaneous pneumomediastinum is characterized by the presence of interstitial air in the mediastinum without any underlying disease. Some cases of spontaneous pneumomediastinum have been reported in the past, although only few reports are available, and its management remains uncertain. This study reviewed our experience in the diagnosis and treatment of spontaneous pneumomediastinum. A retrospective study of 71 cases treated for spontaneous pneumomediastinum at the Yodogawa Christian Hospital between April 2005 and March 2020 was conducted. The patients' mean age was 19.3 years (range, 7-48 years). A triggering event was noted in 69% of the cases. Seventy-six percent of the patients were admitted to the hospital, and 24% were outpatients. Treatment included analgesia, rest, antibiotics, and/or oxygen therapy. Thirty-six patients (51%) were treated with antibiotics. None of the cases presented any complications, including mediastinitis or worsening respiratory condition. Two patients (3%) had a recurrence of spontaneous pneumomediastinum. All the patients, with or without antimicrobial treatment and hospitalization, had favorable outcomes. We should therefore reconsider the need for hospitalization and antimicrobial therapy for patients with mediastinitis prophylaxis.


Sensitivity of Self-Reported Human Papillomavirus Vaccination History Among 18- to 26-Year-Old Men Who Have Sex With Men: Seattle, WA, 2016 to 2018.
We assessed the sensitivity of self-reported human papillomavirus (HPV) vaccination among young adult men who have sex with men (MSM) with documented HPV vaccination. During 2016-2018, MSM and transgender women aged 18 to 26 years were enrolled in Seattle, WA. A history of HPV vaccination was assessed via self-administered survey, clinic electronic medical records, and the Washington State Immunization Information System. We assessed self-report sensitivity among participants with documented prior HPV vaccination (≥1 dose) in either the electronic medical record or the Washington State Immunization Information System, and used logistic regression to compare sensitivity by age, number of doses, and time since first dose. Of 292 participants with ≥1 documented HPV vaccine dose, 243 self-reported ≥1 dose (sensitivity, 83.2%; 95% confidence interval [CI], 78.4%-87.3%). Compared with participants whose first dose was <1 year ago, the likelihood of self-report was lower among those with ≥3 years since first dose (adjusted odds ratio [aOR], 0.2; 95% CI, 0.1-0.5). Furthermore, compared with participants with only 1 documented HPV vaccine dose, the likelihood of self-reporting ≥1 dose was higher among those with 2 (aOR, 2.4; 95% CI, 1.0-5.5) or ≥3 doses (aOR, 6.2; 95% CI, 2.7-14.4). Among 115 participants with ≥3 documented doses, sensitivity for recalling ≥3 doses was 69.6% (95% CI, 60.3%-77.8%). Most young adult MSM with a documented history of HPV vaccination self-reported prior HPV vaccination. Although recall was highest in those with ≥3 doses, 30% of this fully vaccinated subgroup did not correctly recall the number of doses received, highlighting limitations of self-reporting. Furthermore, results indicating reduced recall with ≥3 years since first dose suggest that sensitivity of self-report among young adult MSM may decline over time as adolescent vaccination coverage increases.


Prescription of Sulphonylureas among Patients with Type 2 Diabetes Mellitus in Italy: Results from the Retrospective, Observational Multicentre Cross-Sectional SUSCIPE (Sulphonyl_UreaS_Correct_Internal_Prescription_Evaluation) Study.
Recent guidelines for the treatment of type 2 diabetes mellitus (T2DM) provide evidence supporting limited use of sulphonylureas (SUs), especially in specific risk patient categories, yet data from national registries still suggest their widespread use. The aim of this study was to investigate characteristics of patients with diabetes treated with SUs and quantify the proportion of patients that met the recommendations for use of SUs by recent guidelines and of those presenting characteristics representing an inappropriate prescription risk (IPR). A multicenter, retrospective, cross-sectional, observational study in patients with T2DM receiving treatment with SUs (as monotherapy or in combination with another diabetes therapy) was conducted between 2017 and 2018 in 22 outpatient diabetes clinics across Italy. Exclusion criteria were type 1 diabetes, diabetes mellitus secondary to other conditions, and presence of severe/life-threatening diseases. A total of 510 patients with T2DM (306 men, 204 women; mean age ± standard deviation 69.8 ± 9.3 years) who were receiving treatment with a SU (as monotherapy or in combination therapy) were assessed in the study. Overall, 70.6% [n = 360; 95% confidence interval (CI) 66.4%, 74.5%] were assessed to have an IPR. Of these, approximately half presented one factor for risk of inappropriate prescription, and 27 and 10.6% presented two and three factors, respectively. In terms of factors contributing to the total burden of risk of inappropriate treatment with SUs, 37.5% (95% CI 33.2%, 41.8%) of all patients were obese; 33.3% (95% CI 29.3%, 37.6%)] were aged ≥ 75 years; 18.6% (95% CI 15.3%, 22.3%) had a history of cardiovascular disease; 14.1% (95% CI 11.2%, 17.4%) had chronic renal insufficiency; 1.8% (95% CI 0.8%, 3.3%) had a history of severe hypoglycemia; 1.8% (95% CI 0.8%; 3.3%) had cognitive impairment; and 2.4% (95% CI 1.2%, 4.1%) had a risky occupation. The results of this study provide evidence of a high rate of inappropriate SU prescription risk among patients with T2DM, especially among those with overweight/obesity, older age, history of cardiovascular disease, and hypoglycemia.


[Verapamil intoxication: beware of the delayed effect].
Intoxication with calcium channel inhibitory drugs is rare but mortality rates reach 10 %. We report the case of a 5-year-old girl who had ingested five 240-mg tablets of extended-release verapamil (VLP) and a tablet of bromazepam. Thirty hours after the ingestion she had a vasoplegic shock, heart conduction disorders, and metabolic complications. She was treated in pediatric intensive care with continuous epinephrine and insulin and recovered completely 60h after the ingestion. This case underlines the danger of calcium channel blocker overdose, increased by the extended-release mechanism: the drug effect is to slow down gastric motility - which explains the latency of the symptoms - but this also increases the drug absorption, inducing a vicious circle. These mechanisms in calcium blocker intoxication need to be kept in mind in order to decrease the mortality of such accidents.


Traditional medicine users in a treated chronic disease population: a cross-sectional study in Indonesia.
Traditional medicine (TM) is commonly used as a treatment in Indonesia. This raises the need for an analysis of its potential development and irrational use. Therefore, we analyze the proportion of TM users among chronic disease patients and its associated characteristics to optimize the use of TM in Indonesia. A cross-sectional study of treated adult chronic disease patients was conducted using the fifth Indonesian Family Life Survey (IFLS-5) database. Descriptive analysis was used to identify the proportion of TM users, while a multivariate logistic regression was used to analyze their characteristics. This study included 4901 subjects and identified 27.1% as TM users. The highest TM use was in subjects with cancer (43.9%), liver issues (38.3%), cholesterol issues (34.3%), diabetes (33.6%), and stroke (31.7%). Characteristics associated with TM users were a perception of one's current health as unhealthy (OR 2.59, 95% CI 1.76-3.81), low medication adherence (OR 2.49, 95% CI 2.17-2.85), age above 65 years (OR 2.17, 95% CI 1.63-2.90), having higher education (OR 1.64, 95% CI 1.17-2.29), and residence outside of Java (OR 1.27, 95% CI 1.11-1.45). Low medication adherence among TM users highlights the potentially irrational use of treatment in chronic diseases. Nevertheless, the longstanding use of TM users indicates the potential for its development. Further studies and interventions are needed to optimize TM use in Indonesia.


Effect of dam's diet on PCB accumulation in nursing Osborne Mendel pups.
Upon parturition, dams were fed high fat or high carbohydrate diets to which 1, 10, and 30 ppm of Aroclor 1254 were added to study the effect of diet on fat and PCB accumulation in nursing pups. The percentage of body fat in the carcasses of 0, 8, and 16 day old pups ranged from 2.2 to 18.2% and from 1.2 to 12.0% for animals suckled by dams fed high fat and high carbohydrate rations, respectively. Accumulation of Aroclor 1254 expressed as ppm wet weight or as total PCB's per pup was similar for animals nursed by dams fed either the high fat or high carbohydrate ration, but pups from dams fed a high carbohydrate ration also had significantly less body fat. Therefore, transfer of PCBs via the milk appears to be more a function of the amounts of PCB's consumed by the dam than the diet's ability to increase the percentage of body fat in the pups.


Stereology and morphometry of lung tissue.
This chapter deals with the stereological quantification of structural characteristics of the lung. The aim of design-based stereological methods is the unbiased and efficient estimation of structural features without making any assumptions on the underlying nature of the biological sample. The methods are based on rigorous sampling of location and orientation, the application of appropriate test systems, and the controlling of the precision of the estimates. Here, we describe the workflow from the fixation of the lung over the processing of the tissue samples to gaining estimates on the structural properties of the lung. Specifically, this chapter deals with methods for estimating the reference volume, sampling location, and sampling orientation, estimating volumes and surface areas of alveolar compartments, estimating total alveolar number, performing stereology at light and electron microscopic level, and dealing with technical problems such as tissue shrinkage. The procedures are illustrated using a worked example from the authors' own laboratory.


In vitro association of fragments of a beta-sheet membrane protein.
Although the beta-barrel membrane protein OmpA can be produced in a biologically active form in E. coli from co-expressed fragments, the fragments have not been demonstrated to associate in vitro. We have produced 3 complementary fragment pairs of OmpA which can associate to form a folded complex according to the SDS band-shift assay. We are able to convert 25-35% of the fragment populations to non-covalent but SDS-stable complexes. The periplasmic chaperone Skp effectively prevented this association. Two separately expressed and purified overlapping fragments of OmpA can form a protease-resistant complex that undergoes the characteristic band-shift upon heating. Our work demonstrates that although membrane insertion and folding of beta-barrel membrane proteins may be a cooperative process, the fragments can associate in vitro without any additional components. However, the low yield and slow folding rates indicate that partially unfolded or destabilized beta-sheet membrane proteins can potentially engage in many non-native interactions.


Overexpression of GRAS Rhizomucor miehei lipase in Yarrowia lipolytica via optimizing promoter, gene dosage and fermentation parameters.
Rhizomucor miehei lipase (RML), a GRAS catalyst with wide applications, was overexpressed in Yarrowia lipolytica, also a GRAS unconventional yeast, via a combined strategy, optimization for promoter, gene dosage and fermentation process. The lipase activity of the recombinant strain was first increased from 19.5 to 26.9 U/mL via codon optimization of rml gene. Subsequently, a method was developed for constructing hybrid promoters harboring different copy number of upstream activation sequences fragment (UAS1B), and the recombinant strain Po1g/hp12d-rml 25# reached 38.9 U/mL. On this basis, expression vectors with different optimized rml gene copy numbers were constructed and introduced into Y. lipolytica Po1g. The recombinant strain Po1g/hp12d-2rml 14# carrying 12 copies of UAS1B in the upstream of pLEUmin and 2 copies of rml gene obtained the highest lipase activity of 59.6 U/mL. Moreover, in optimized shaking flask culture parameters: 5% (m/v) of d-Sorbitol, 2% (v/v) inoculation density, initial pH 7.0, and 30 mL initial culture medium, the RML activity of Po1g/hp12d-2rml 14# further reached 157 U/mL after 84-h of incubation at 28 ℃. Overall, RML activity was enhanced about 8-fold compared with the initial recombinant strain via the combined strategy, which provides a consolidated basis for the large-scale production of RML in Y. lipolytica to match urgent demand of the market.


Reverse Micelle Strategy for the Synthesis of MnO x -TiO2 Active Catalysts for NH3-Selective Catalytic Reduction of NO x at Both Low Temperature and Low Mn Content.
MnO x -TiO2 catalysts (0, 1, 5, and 10 wt % Mn nominal content) for NH3-SCR (selective catalytic reduction) of NO x have been synthesized by the reverse micelle-assisted sol-gel procedure, with the aim of improving the dispersion of the active phase, usually poor when obtained by other synthesis methods (e.g., impregnation) and thereby lowering its amount. For comparison, a sample at nominal 10 wt % Mn was obtained by impregnation of the (undoped) TiO2 sample. The catalysts were characterized by using an integrated multitechnique approach, encompassing X-ray diffraction followed by Rietveld refinement, micro-Raman spectroscopy, N2 isotherm measurement at -196 °C, energy-dispersive X-ray analysis, diffuse reflectance UV-vis spectroscopy, temperature-programmed reduction technique, and X-ray photoelectron spectroscopy. The obtained results prove that the reverse micelle sol-gel approach allowed for enhancing the catalytic activity, in that the catalysts were active in a broad temperature range at a substantially low Mn loading, as compared to the impregnated catalyst. Particularly, the 5 wt % Mn catalyst showed the best NH3-SCR activity in terms of both NO x conversion (ca. 90%) and the amount of produced N2O (ca. 50 ppm) in the 200-250 °C temperature range.


Early prediction of renal graft function: Analysis of a multi-center, multi-level data set.
Long-term graft survival rates after renal transplantation are still poor. We aimed to build an early predictor of an established long-term outcomes marker, the estimated glomerular filtration rate (eGFR) one year post-transplant (eGFR-1y). A large cohort of 376 patients was characterized for a multi-level bio-marker panel including gene expression, cytokines, metabolomics and antibody reactivity profiles. Almost one thousand samples from the pre-transplant and early post-transplant period were analysed and employed for machine learning-assisted prediction. Pre-transplant data led to a prediction achieving a Pearson's correlation coefficient of r=0.38 between measured and predicted eGFR-1y. Two weeks post-transplant, the correlation was improved to r=0.63, and at the third month, to r=0.76. eGFR values were stable throughout the first post-transplant year. Several characteristics were predictive for eGFR, including age of donor and recipient, body mass index, HLA mismatch, cytomegalovirus mismatch and valganciclovir prophylaxis. Additionally, a subset of 19 nuclear magnetic resonance bins of the urine metabolome data was shown to have potential applications in non-invasive eGFR monitoring. Importantly, we identified the expression of the genes TMEM176B and HMMR as potential prognostic markers for changes in the eGFR after the second post-transplantation week. Our multi-center, multi-level data set represents a milestone in the efforts to predict transplant outcome. While an acceptable predictive capacity was achieved, we are still far from predicting changes in the eGFR precisely. Additional studies employing further marker panels are needed to establish predictors of eGFR-1y for clinical application; herein, gene expression markers seem to hold the most promise.


The rising root sign: the magnetic resonance appearances of post-operative spinal subdural extra-arachnoid collections.
We present a case series of symptomatic post-operative spinal subdural extra-arachnoid collections that displace the cauda equina roots anteriorly. This is described as the "rising root sign".


Adverse events occurring post-covid-19 vaccination among healthcare professionals - A mixed method study.
Healthcare professionals (HCPs) are at the front line of the nation's fight against COVID-19 and are always at a greater risk of contracting contagious disease. But amidst the crisis, the vaccines were not accepted by all the HCPs due to adverse events occurring post-COVID-19 vaccination. Hence, the present study was designed to assess adverse events occurring among HCPs post-COVID-19 vaccination both quantitatively and qualitatively. Sequential mixed-method approach was employed. A cross-sectional E-survey was conducted among the healthcare professionals of a North Indian (*Statistically significant (p < 0.05)) college and hospital. The second phase included a semi-structured qualitative interview of the participants who were willing to participate. Among all the HCPs with age groups ranging from 20 to 70 years, majority of them experienced pain at the site of injection (88.8-100%) followed by tiredness (87.7-60%) and body ache (86.6-40%) post-vaccination. There is an increased frequency of adverse events in females as compared to males. Qualitative findings are summarised in three major domains i.e vaccine adverse effects, fear and hesitancy for vaccines and vaccine acceptance. Short term adverse events of COVISHIELD vaccine were very few and were mild in severity yet interviews showed hesitancy of study participants for vaccination.


Assessment of CareStart G6PD rapid diagnostic test and CareStart G6PD biosensor in Mauritania.
The elimination of Plasmodium vivax malaria requires 8-aminoquinolines, which are contraindicated in patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency due to the risk of acute haemolytic anaemia. Several point-of-care devices have been developed to detect G6PD deficiency. The objective of the present study was to evaluate the performance of two of these devices against G6PD genotypes in Mauritania. Outpatients were screened for G6PD deficiency using CareStart™ rapid diagnostic test (RDT) and CareStart™ G6PD biosensor in Nouakchott, Mauritania, in 2019-2020. African-type and Mediterranean-type G6PD genotypes commonly observed in Africa were determined by polymerase chain reaction-restriction fragment length polymorphism and sequencing. Qualitative variables were compared using Fisher's exact test. Of 323 patients (74 males and 249 females), 5 males and 2 homozygous females had the African-type A- genotype: A-(202) in 3 males and 2 females and G6PD A-(968) in 2 males. Among heterozygous females, 13 carried G6PD A-(202), 12 G6PD A-(968), and 3 G6PD A-(542) variants. None had the Mediterranean-type G6PD genotype. Eight had a positive G6PD RDT result, including all 7 hemizygous males and homozygous females with A- or A-A- (0.12 to 2.34 IU/g haemoglobin, according to G6PD biosensor), but RDT performed poorly (sensitivity, 11.1% at the cut-off level of < 30%) and yielded many false negative tests. Thirty-seven (50.0%) males and 141 (56.6%) females were anaemic. The adjusted median values of G6PD activity were 5.72 and 5.34 IU/g haemoglobin in non-anaemic males (n = 35) and non-anaemic males and females (n = 130) with normal G6PD genotypes using G6PD biosensor, respectively. Based on the adjusted median of 5.34 IU/g haemoglobin, the performance of G6PD biosensor against genotyping was as follows: at 30% cut-off, the sensitivity and specificity were 85.7% and 91.7%, respectively, and at 80% cut-off, the sensitivity was 100% while the specificity was 64.9%. Although this pilot study supports the utility of biosensor to screen for G6PD deficiency in patients, further investigation in parallel with spectrophotometry is required to promote and validate a more extensive use of this point-of-care device in areas where P. vivax is highly prevalent in Mauritania.


The polymorphisms of LCR, E6, and E7 of HPV-58 isolates in Yunnan, Southwest China.
Variations in HPV LCR/E6/E7 have been shown to be associated with the viral persistence and cervical cancer development. So far, there are few reports about the polymorphisms of the HPV-58 LCR/E6/E7 sequences in Southwest China. This study aims to characterize the gene polymorphisms of the HPV-58 LCR/E6/E7 sequences in women of Southwest China, and assess the effects of variations on the immune recognition of viral E6 and E7 antigens. Twelve LCR/E6/E7 of the HPV-58 isolates were amplified and sequenced. A neighbor-joining phylogenetic tree was constructed by MEGA 7.0, followed by the secondary structure prediction of the related proteins using PSIPRED v3.3. The selection pressure acting on the HPV-58 E6 and E7 coding regions was estimated by Bayes empirical Bayes analysis of PAML 4.8. Meanwhile, the MHC class-I and II binding peptides were predicted by the ProPred-I server and ProPred server. The transcription factor binding sites in the HPV-58 LCR were analyzed using the JASPAR database. Twenty nine SNPs (20 in the LCR, 3 in the E6, 6 in the E7) were identified at 27 nucleotide sites across the HPV-58 LCR/E6/E7. From the most variable to the least variable, the nucleotide variations were LCR > E7 > E6. The combinations of all the SNPs resulted in 11 unique sequences, which were clustered into the A lineage (7 belong to A1, 2 belong to A2, and 2 belong to A3). An insertion (TGTCAGTTTCCT) was found between the nucleotide sites 7280 and 7281 in 2 variants, and a deletion (TTTAT) was found between 7429 and 7433 in 1 variant. The most common non-synonymous substitution V77A in the E7 was observed in the sequences encoding the α-helix. 63G in the E7 was determined to be the only one positively selected site in the HPV-58 E6/E7 sequences. Six non-synonymous amino acid substitutions (including S71F and K93 N in the E6, and T20I, G41R, G63S/D, and V77A in the E7) were affecting multiple putative epitopes for both CD4+ and CD8+ T-cells. In the LCR, C7265G and C7266T were the most variable sites and were the potential binding sites for the transcription factor SOX10. These results provide an insight into the intrinsic geographical relatedness and biological differences of the HPV-58 variants, and contribute to further research on the HPV-58 epidemiology, carcinogenesis, and therapeutic vaccine development.


Properties of the promoter region of the T18d (T13c) Tla gene.
The T18d (formerly T13c) gene of BALB/c mice belongs to the category of Tla genes which is expressed by both thymocytes and TL+ T-cell leukemias. To elucidate the regulation of T18d, different restriction fragments of the 5' flanking region between -457 and +146 were linked to the chloramphenicol acetyltransferase (CAT) gene and transfected into TL+ and TL- cells. By comparison of transiently expressed CAT activity among cells transfected with different CAT constructs, the results suggest that determination of TL+ vs TL- phenotypes is located within the region -105 to -33, and that an element essential to T18d expression resides within the region -33 to +54. Putative DNA-binding factors characterizing particular cell types and displaying selective affinity for particular T18d restriction fragments were identified by electrophoretic mobility shift assays with nuclear extracts (NEs). Two factors (or complexes) which bound specifically to the T18d fragment -105 to -33 were expressed preferentially in TL+ cells and thus may be involved in determining the tissue-selective expression of T18d. The close proximity of negative and positive cis-acting elements within the promoter region is consistent with regulation of T18d gene expression by a variety of trans-acting factors whose production is attuned to development and differentiation. The data provided may serve as a guide to study the regulation of other categories of Tla genes that are normally silent in thymocytes but may become expressed by leukemia cells.


H3 ubiquitination by NEDD4 regulates H3 acetylation and tumorigenesis.
Dynamic changes in histone modifications under various physiological cues play important roles in gene transcription and cancer. Identification of new histone marks critical for cancer development is of particular importance. Here we show that, in a glucose-dependent manner, E3 ubiquitin ligase NEDD4 ubiquitinates histone H3 on lysine 23/36/37 residues, which specifically recruits histone acetyltransferase GCN5 for subsequent H3 acetylation. Genome-wide analysis of chromatin immunoprecipitation followed by sequencing reveals that NEDD4 regulates glucose-induced H3 K9 acetylation at transcription starting site and enhancer regions. Integrative analysis of ChIP-seq and microarray data sets also reveals a consistent role of NEDD4 in transcription activation and H3 K9 acetylation in response to glucose. Functionally, we show that NEDD4-mediated H3 ubiquitination, by transcriptionally activating IL1α, IL1β and GCLM, is important for tumour sphere formation. Together, our study reveals the mechanism for glucose-induced transcriptome reprograming and epigenetic regulation in cancer by inducing NEDD4-dependent H3 ubiquitination.


The translatability of genetic networks from model to crop species: lessons from the past and perspectives for the future.
Comparative analyses of growth-regulatory mechanisms between Arabidopsis and maize revealed that even when the gene space is conserved, the translation of knowledge from model species to crops is not trivial. Based on these insights, we formulate future opportunities to improve the interpretation of curiosity-driven research towards crop improvement.


Validation of the liver traffic light test as a predictive model for survival and development of liver-related events.
Liver disease mortality rates continue to rise due to late diagnosis. We need noninvasive tests to be made available in the community that can identify patients at risk from a serious liver-related event (SLE). We examine the performance of a blood test, the liver traffic light test (LTLT), with regard to its ability to predict survival and SLEs. Using routinely gathered clinical data, sequential LTLT test results from 4854 individuals with suspected liver disease were prospectively analyzed (median follow-up 41 months). An SLE was defined as the development of cirrhosis, liver failure, ascites, or varices. Patients were graded as follows: red (high risk), amber (intermediate risk), and green (low risk). Overall, 565 individuals experienced an SLE (11.6%). The area under the curve (AUC) for the continuous LTLT variable was 0.87 (95% confidence interval 0.85-0.89) for prediction of an SLE and 0.81 (0.78-0.84) for mortality. When categorized into red/amber/green grades, a red LTLT result predicted an SLE with negative and positive predictive values of 0.97 and 0.29, respectively. A red LTLT score predicted mortality with negative and positive predictive values of 0.98 and 0.18, respectively. Kaplan-Meier plots demonstrated increased mortality and SLEs in the red group versus the green and amber groups (P < 0.001) and an increase in SLEs in the amber versus green group (P < 0.001). Here, the LTLT is further validated for the prediction of survival and SLE development. The LTLT could aid primary care risk management and referral pathways with the aim of detecting and treating liver disease earlier in the general population.


Eradication of Helicobacter pylori: Past, present and future.
Helicobacter pylori is the major cause of chronic gastritis and peptic ulcers. Since the classification as a group 1 carcinogenic by International Agency for Research on Cancer, the importance of the complete H. pylori eradication has obtained a novel meaning. Hence, several studies have been made in order to deepen the knowledge in therapy strategies. However, the current therapy presents unsatisfactory eradication rates due to the lack of therapeutic compliance, antibiotic resistance, the degradation of antibiotics at gastric pH and their insufficient residence time in the stomach. Novel approaches have been made in order to overcome these limitations. The purpose of this review is to provide an overview about the current therapy and its limitations, while highlighting the possibility of using micro- and nanotechnology to develop gastric drug delivery systems, overcoming these difficulties in the future.


Characterization of amino acid variation at strategic positions in parasite and human proteases for selective inhibition of falcipains in Plasmodium falciparum.
Falcipains (FP) of Plasmodium falciparum are important virulence factors marked as potential targets for antimalarial drug discovery. In this study, the previously uncharacterized fp2B (PF11_0161) was shown to be highly expressed as an active enzyme during the erythrocytic stage. With three related proteases in the FP family and the existence of human homologues, it is prudent to identify clusters of residues unique to the parasite proteases that can be targeted selectively for drug design. Using bioinformatic tools, we have carefully mapped out a highly conserved and unique region constituted by I85, S149, and A151 in the plasmodial proteases that can influence the development of compounds capable of inhibiting the entire FP family. Taking drug interactions with the human homologues into consideration, these residues in FP2B were replaced with the cognate residues found in human cathepsin L (catL) for evaluation. Despite the high sequence similarity between the FP2 isozymes (97.5%), FP2B is found to be more tolerant to amino acid substitution at position 149 than FP2A. This structural disparity implied that residues mediating peptide substrate interactions are not fully conserved across the FP family and warrant attention in the design and evaluation of protease inhibitors focused on the FPs. The simultaneous substitution of the neighboring residues (I85 or A151) rendered the double mutants (S149A/I85M and S149A/A151D) completely inactive. Significantly, the mutations did not result in 'catL-like' specificity, suggesting that substrate-based inhibitors could be rationally designed against these important parasite-specific structural determinants.


Translation and psychometric properties of the Chinese (Mandarin) version of the Child Oral Health Impact Profile-Short Form 19 (COHIP-SF 19) for school-age children.
Although caries and malocclusion occur with a high prevalence in Chinese school-age children, there were no appropriate instrument to assess the oral health-related quality of life (OHRQoL) for this population. The aim of our study was to develop a Chinese (Mandarin) version of the Child Oral Health Impact Profile-Short Form 19 (COHIP-SF 19) and provide a preliminary test of its psychometric properties. The Chinese version of COHIP-SF 19 was developed through a standard translation and back translation procedure. The psychometric properties of the instrument were tested among 644 school-age children in Beijing, China, including the internal consistency, test-retest reliability, discriminant and convergent validity. A Mann-Whitney U test was used to determine the capability of the instrument to differentiate children with different caries and malocclusion outcomes. And partial Spearman correlations were used to determine the relationships between the OHRQoL scores and clinical-severity indicators and self-perceived health ratings, respectively. Chinese school-age children had relatively high OHRQoL scores, in spite of the fact that oral impacts were quite common (56.3%). The internal consistency and retest reliability were good to excellent with a Chronbach's alpha of 0.81 and an intra-class correlation coefficient (ICC) of 0.77. Children who had active tooth decay or severe malocclusion had significantly lower COHIP-SF 19 scores (P ≤0.001). Girls had somewhat higher scores in the oral health and functional well-being subscales (P <0.05), while children from rural districts had lower scores than children from urban areas (P <0.05). We observed a low to moderate correlation between the overall COHIP-SF 19, subscale scores and clinical severity indicators as well as self-perceived health ratings, after adjustment for children's age, gender, and school district (│r s │ =0.11 - 0.51, P <0.05). We confirmed satisfactory psychometric properties for the Chinese version of COHIP-SF 19 in a community sample of Chinese school-age children. The OHRQoL instrument should play a more important role in future clinical studies, epidemiological surveys and potential public health policy in China.


Which chemicals should be grouped together for mixture risk assessments of male reproductive disorders?
There is concern about cumulative exposures to compounds that disrupt male sexual differentiation in foetal life, leading to irreversible effects in adulthood, including declines in semen quality, testes non-descent, malformations of the penis and testis cancer. Traditional chemical-by-chemical risk assessment approaches cannot capture the likely cumulative health risks. Past efforts of focusing on combinations of phthalates, a subgroup of chemicals suspected of contributing to these risks, do not go far enough, as they ignore the contribution of other types of chemicals. With the aim of providing criteria for the inclusion of additional chemicals in mixture risks assessments for male reproductive health, this paper examines the mechanisms of action of various chemicals capable of disrupting male sexual differentiation. An Adverse Outcome Pathway (AOP) network for malformations of the male reproductive system is constructed that includes new findings about the role of disruptions of prostaglandin signalling. This network is used to identify pathways that converge at critical nodal points to produce down-stream adverse effects. From this knowledge, combinations of chemicals with different mechanisms of action are predicted that should result in cumulative effects. These predictions are then mapped against evidence from experimental mixture studies with relevant combinations. From the outcome of this analysis it is concluded that cumulative assessment groups for male reproductive health risks should not only include phthalates but also comprise androgen receptor (AR) antagonists, chemicals capable of disrupting steroid synthesis, InsL3 production, prostaglandin signalling and co-planar polychlorinated dibenzo-dioxins together with other dioxin-like compounds. This list goes far beyond what has been suggested previously. A minimum set of chemicals to be assessed together with phthalates includes pesticides such as vinclozolin, prochloraz, procymidone, linuron, the pain killers paracetamol, aspirin and ibuprofen, pharmaceuticals such as finasteride, ketoconazole, and the lipid-lowering drug simvastin, poly-chlorinated dibenzo-dioxins and other dioxin-like pollutants and phenolics such as bisphenol A and butylparaben. AOP network analyses are essential to overcome difficulties in establishing groupings of chemicals for mixture risk assessments that derive from a narrow focus on mechanisms and modes of action.


Comparison of four standards for determining adequate water intake of nursing home residents.
Adequate hydration for nursing home residents is problematic. The purpose of this study was to compare four standards used to determine a recommended water intake among nursing home residents. Inconsistencies in the amount of water intake recommended based on the standards compared were identified. The standard based on height and weight provides the most individualized recommendation. An individualized recommendation would facilitate goal setting for the care plan of each older person and assist in the prevention of dehydration. It is essential that a cost-effective and clinically feasible approach to determine adequate water intake be determined for this population to prevent the adverse outcomes associated with dehydration.


Wnt signaling in gut organogenesis.
Wnt signaling regulates some aspect of development of nearly all endoderm-derived organs and Wnts mediate both differentiation and proliferation at different steps during visceral organogenesis. Wnt2b induces liver formation in zebrafish 1 and may combine with other inducers, Fibroblast Growth Factors 1 & 4 and Bone Morphogenetic Protein 4, to specify the mammalian liver.2-5 Later in development, Wnts are critical for liver expansion and, finally, for terminal hepatocyte differentiation,6-12 as reviewed elsewhere in this issue (Monga). Likewise, in the pancreas, Wnts drive proliferation of exocrine and endocrine cells13,14 and promote acinar cell differentiation,13,15 as reviewed in the chapter by Murtaugh. Here we examine the intricate involvement of Wnt signaling in growth and differentiation of the digestive tract.


A common genetic variant in fatty acid amide hydrolase is linked to alterations in fear extinction neural circuitry in a racially diverse, nonclinical sample of adults.
Poor fear extinction learning and recall are linked to the development of fear-based disorders, like posttraumatic stress disorder, and are associated with aberrant activation of fear-related neural circuitry. This includes greater amygdala activation during extinction learning and lesser hippocampal and ventromedial prefrontal cortex (vmPFC) activation during recall. Emerging data indicate that genetic variation in fatty acid amide hydrolase (FAAH C385A; rs324420) is associated with increased peripheral endocannabinoid (eCB) levels and lesser threat-related amygdala reactivity. Preclinical studies link increased eCB signaling to better extinction learning and recall, thus FAAH C385A may protect against the development of trauma-related psychopathology by facilitating extinction learning. However, how this FAAH variant affects fear extinction neural circuitry remains unknown. In the present study, we used a novel, immersive-reality fear extinction paradigm paired with functional neuroimaging to assess FAAH C385A effects on fear-related neural circuitry and conditioned fear responding (US expectancy ratings, subjective units of distress, and skin conductance responding) in healthy adults from an urban area (Detroit, MI; N = 59; C/C = 35, A-carrier = 24). We found lesser amygdala activation in A-allele carriers, compared to C/C homozygotes, during early extinction recall. Likewise, we found lesser dorsal anterior cingulate cortex and greater hippocampus activation in early extinction learning in A-carriers compared to C/C homozygotes. We found no effects of FAAH C385A on vmPFC activation or behavioral fear indices. These data support and extend previous findings that FAAH genetic variation, associated with increased eCB signaling and subsequent enhanced fear extinction, may predict individual differences in successful fear learning.


Left ventricular assist device implantation via lateral thoracotomy: A systematic review and meta-analysis.
Left ventricular assist device (LVAD) implantation via lateral thoracotomy can offer similar effectiveness to conventional approaches with less perioperative adverse events. We performed a systematic review and meta-analysis to determine the potential benefits of lateral thoracotomy (LT) for LVAD implantation compared to median sternotomy. We searched MEDLINE and Embase databases for studies comparing continuous-flow LVAD implantation using LT with conventional sternotomy. Main outcomes were perioperative mortality and complications. Twenty-five observational studies enrolling 3072 patients were included with a median follow-up of 10 months. Perioperative mortality (30 day or in-hospital) was 7% (LT) and 14% (sternotomy); however, mortality differences were no longer statistically significant in matched/adjusted studies (RR:0.86; 95%CI:0.52-1.44; p = 0.58). LT was associated with decreased need for blood product transfusions (mean difference[MD]: -4.7; 95%CI: -7.2 to -2.3 units; p < 0.001), reoperation for bleeding (RR:0.34; 95%CI:0.22-0.54; p < 0.001), postoperative RVAD implantation (RR:0.53; 95%CI:0.36-0.77; p < 0.001), days requiring inotropes (MD: -1.1; 95%CI: -2.1 to -0.03 inotrope days; p = 0.04), ICU (MD: -3.3; 95%CI: -6.0 to -0.7 ICU days; p = 0.01), and hospital length of stay (MD: -5.1; 95%CI: -10.1 to -0.1 hospital days; p = 0.04) in matched/adjusted studies. Overall mortality during follow-up was significantly lower for LT in unmatched/unadjusted studies but not statistically significantly lower in matched/adjusted studies (Hazard Ratio:0.82; 95%CI:0.59-1.14; p = 0.24). LVAD implantation via LT was associated with significantly decreased need for blood products, reoperation for bleeding, and postoperative RVAD implantation. Furthermore, days on inotropic support were also lower, likely contributing to the shorter length of stay. These findings support greater use of a LT approach for carefully selected patients.


Process development of a recombinant antibody/interleukin-2 fusion protein expressed in protein-free medium by BHK cells.
The production, purification and stability of quality (in terms of integrity and glycosylation) of an antibody/interleukin-2 fusion protein with potential application in tumour-targeted therapy expressed in BHK21 cells are described. Consistency of the product throughout time was determined by analysis of glycosylation of the fusion protein using MALDI-TOF mass spectroscopy and HPAEC-PAD combined with product integrity studies by SDS-PAGE and Western blotting. These investigations showed consistent expression in terms of integrity and of three major oligosaccharide structures of the fusion protein after 62 generations. The data obtained at this stage indicated the suitability of the cell line for production purposes. Different approaches for the production of this protein were subsequently carried out. The relative productivity of the recombinant fusion protein and general performance of the cells in two different protein-free medium (PFM) culture systems, continuous chemostat and continuous perfusion using a Centritech centrifuge as a cell retention device, were studied. The results indicate that the chemostat culture resulted in more stable and controllable nutrient environment, which could indicate better product consistency, in accordance with what has been observed under serum-containing conditions, in relation to the perfusion culture. Finally, product obtained from the chemostat culture was analysed and purified. The purification process was optimised with an increase in the overall yield from 38 to 70% being obtained, a significant improvement with important consequences for the implementation of an industrial-scale culture system. In conclusion, it was possible to produce and purify the recombinant antibody/interleukin-2 fusion protein assuring the quality and stability of the product in terms of integrity and glycosylation. Therefore, a candidate production process was established.


Comparison of tumor size assessments in tumor growth inhibition-overall survival models with second-line colorectal cancer data from the VELOUR study.
To compare lesion-level and volumetric measures of tumor burden with sum of the longest dimensions (SLD) of target lesions on overall survival (OS) predictions using time-to-growth (TTG) as predictor. Tumor burden and OS data from a phase 3 randomized study of second-line FOLFIRI ± aflibercept in metastatic colorectal cancer were available for 918 patients out of 1216 treated (75%). A TGI model that estimates TTG was fit to the longitudinal tumor size data (nonlinear mixed effect modeling) to estimate TTG with: SLD, sum of the measured lesion volumes (SV), individual lesion diameters (ILD), or individual lesion volumes (ILV). A parametric OS model was built with TTG estimates and assessed for prediction of the hazard ratio (HR) for survival. Individual lesions had consistent dynamics within individuals. Between-lesion variability in rate constants was lower (typically < 27% CV) than inter-patient variability (typically > 50% CV). Estimates of TTG were consistent (around 12 weeks) across tumor size assessments. TTG was highly significant in a log-logistic parametric model of OS (median over 12 months). When individual lesions were considered, TTG of the fastest progressing lesions best predicted OS. TTG obtained from the lesion-level analyses were slightly better predictors of OS than estimates from the sums, with ILV marginally better than ILD. All models predicted VELOUR HR equally well and all predicted study success. This analysis revealed consistent TGI profiles across all tumor size assessments considered. TTG predicted VELOUR HR when based on any of the tumor size measures.


Detection of trace anthracene in soil samples with real-time fluorescence quantitative immuno-PCR using a molecular beacon probe.
We developed a highly sensitive and robust real-time fluorescence quantitative immuno-PCR (RTFQ-IPCR) method which uses molecular beacon (MB) probe to detect trace anthracene in the environment. This method was performed on serial dilutions of known anthracene concentrations equivalent to 10-fold dilutions of 10fg/mL to 100pg/mL. We obtained a linear relationship between 10fg/mL and 100pg/mL, with y=0.684x+13.221. A correlation coefficient of 0.994 was also identified, with a detection limit of 4.5fg/mL. After investigating the presence of anthracene in soil samples via RTFQ-IPCR, the obtained concentrations were confirmed by ELISA to be correct and believable, with the recovery ratio ranging from 82% to 112.5%. Based on its sensitivity and reproducibility, MB-based RTFQ-IPCR was found to be acceptable for use in on-site field tests to provide rapid, quantitative, and reliable test results for making environmental decisions.


A Tudor staphylococcal nuclease from Penaeus monodon: cDNA cloning and its involvement in RNA interference.
RNA interference (RNAi) plays an important role in an antiviral defense in shrimp. RNAi technology has been extensively used for inhibition of viral replication and studying gene function. However, the mechanism of shrimp RNAi pathway is still poorly understood. In this study, we identified and characterized an additional protein in the RNAi pathway, Tudor staphylococcal nuclease from Penaeus monodon (PmTSN). The full-length cDNA of PmTSN is 2897 bp, with an open reading frame encoding a putative protein of 889 amino acids. Phylogenetic analysis and domain structure comparison revealed that PmTSN is more closely related to vertebrate TSN by sharing the amino acid sequence identity of 57% with TSN of zebrafish. This represents a new type of TSN proteins by exhibiting the four tandem repeat of staphylococcal nuclease-like domain (SN), followed by a Tudor and a partially truncated C-terminal SN domain. Knockdown of PmTSN by dsRNA targeting SN3 domain resulted in the impairment of dsRNA targeting PmRab7 gene to silence PmRab7 expression. In addition, the efficiency of dsRNA targeting YHV-protease gene inhibiting yellow head virus replication was decreased in the PmTSN-knockdown shrimps. Our results imply that PmTSN is involved in dsRNA-mediated gene silencing in shrimp and thus we identified the additional protein involved in shrimp RNAi pathway.


Glycosylation and cross-linking in bone type I collagen.
Fibrillar type I collagen is the major organic component in bone, providing a stable template for mineralization. During collagen biosynthesis, specific hydroxylysine residues become glycosylated in the form of galactosyl- and glucosylgalactosyl-hydroxylysine. Furthermore, key glycosylated hydroxylysine residues, α1/2-87, are involved in covalent intermolecular cross-linking. Although cross-linking is crucial for the stability and mineralization of collagen, the biological function of glycosylation in cross-linking is not well understood. In this study, we quantitatively characterized glycosylation of non-cross-linked and cross-linked peptides by biochemical and nanoscale liquid chromatography-high resolution tandem mass spectrometric analyses. The results showed that glycosylation of non-cross-linked hydroxylysine is different from that involved in cross-linking. Among the cross-linked species involving α1/2-87, divalent cross-links were glycosylated with both mono- and disaccharides, whereas the mature, trivalent cross-links were primarily monoglycosylated. Markedly diminished diglycosylation in trivalent cross-links at this locus was also confirmed in type II collagen. The data, together with our recent report (Sricholpech, M., Perdivara, I., Yokoyama, M., Nagaoka, H., Terajima, M., Tomer, K. B., and Yamauchi, M. (2012) Lysyl hydroxylase 3-mediated glucosylation in type I collagen: molecular loci and biological significance. J. Biol. Chem. 287, 22998-23009), indicate that the extent and pattern of glycosylation may regulate cross-link maturation in fibrillar collagen.


Time course of induction of a heat shock gene (hsp70) in the rabbit cerebellum after LSD in vivo: involvement of drug-induced hyperthermia.
In situ hybridization studies were carried out to determine whether induction of hsp70 mRNA in various cellular layers of the rabbit cerebellum was due to hyperthermic effects of the psychotropic drug LSD. Results indicated that induction was not present when LSD-induced hyperthermia was blocked. The pattern of induction of hsp70 mRNA in various cell types of the cerebellum was similar when hyperthermia was induced by either drug (LSD) or nondrug means (placement of animals in a warm incubator). A time course analysis of the induction of hsp70 mRNA following LSD-induced hyperthermia revealed maximal levels of mRNA at 1 hr in all cerebellar cell layers except the Purkinje layer where highest levels were attained at 5 hr. By 10 hr hsp70 mRNA had returned to constitutive levels in all cellular layers of the cerebellum.


Efficiency and sensitivity of indirect immunoperoxidase methods.
The peroxidase-antiperoxidase (PAP) complex method has repeatedly been claimed to be more sensitive and antibody efficient than the indirect peroxidase labeled antibody method. However, most studies comparing these methods used tissue sections as the test material. However, test systems with known amounts of antigen will allow more reliable comparison of these methods and quantitative evaluation of method sensitivity. We therefore compared the antibody efficiency and sensitivity of these methods for the detection of human chorionic gonadotropin in an enzyme linked immunosorbent assay (ELISA), an antigen spot test (AST) and tissue sections of choriocarcinoma. In the PAP technique rabbit PAP and goat anti-rabbit antibody were applied. The same antibody was peroxidase-labeled with the periodate technique and used in the labeled antibody method. In the ELISA the PAP method resulted in slightly higher antibody efficiency than the labeled antibody method. At low primary antibody dilutions the intensity of the reaction decreased with the PAP method but remained high with the labeled antibody method, in the ELISA as well as on tissue sections. In the AST the labeled antibody method and the PAP method appeared to be equally sensitive.


Short and medium term outcomes after rollerball endometrial ablation for menorrhagia.
To review prospectively the intraoperative, short and medium term outcomes of patients treated by rollerball endometrial ablation during the learning curve for this relatively new procedure. Seventy-seven women with menorrhagia (71 with dysfunctional bleeding; six with additional small intramural myomas) underwent hysteroscopic rollerball endometrial ablation by coagulation diathermy during 1.5% glycine irrigation, after a two-month period of hormonal suppression. OPERATIVE AND SHORT-TERM EFFECTS: These were recorded at operation and six-week follow-up. No serious short-term complications were recorded. Objective measurement of intraoperative blood loss was always less than 20 mL. One woman experienced uterine perforation with a narrow cervical dilator which did not interfere with the ablation, and two patients had mild postoperative uterine infections. One patient experienced persistent postoperative pain. MEDIUM-TERM OUTCOME: This was recorded at each visit, or by telephone, and confirmed by questionnaire at one year. Twenty-five per cent achieved complete amenorrhoea, 29% staining only, 30% light periods, 10% "normal" or erratic periods and 6% were unchanged. Five patients underwent a second ablation, and three of these later underwent hysterectomy. Measured menstrual blood loss fell from 104 +/- 19 mL (mean +/- standard error of mean) to 1.7 +/- 1.1 mL at six months in 18 women. Other menstrual symptoms were also often dramatically reduced. Of those women with dysmenorrhoea, 33% were cured and 43% markedly improved; of those with midcycle pain, 28% were cured and 53% markedly improved; of those with significant premenstrual symptoms, 13% were cured, 47% markedly improved, 11% unchanged, and 6% were worse. This new procedure is a safe and effective treatment for menorrhagia caused by dysfunctional uterine bleeding, with impressive ancillary benefits for dysmenorrhoea, midcycle pain and premenstrual tension. It is often called minimally invasive surgery, but it must be recognised that it is not minor surgery.


Myelinating and demyelinating phenotype of Trembler-J mouse (a model of Charcot-Marie-Tooth human disease) analyzed by atomic force microscopy and confocal microscopy.
The accumulation of misfolded proteins is associated with various neurodegenerative conditions. Mutations in PMP-22 are associated with the human peripheral neuropathy, Charcot-Marie-Tooth Type 1A (CMT1A). PMP-22 is a short-lived 22 kDa glycoprotein, which plays a key role in the maintenance of myelin structure and compaction, highly expressed by Schwann cells. It forms aggregates when the proteasome is inhibited or the protein is mutated. This study reports the application of atomic force microscopy (AFM) as a detector of profound topographical and mechanical changes in Trembler-J mouse (CMT1A animal model). AFM images showed topographical differences in the extracellular matrix and basal lamina organization of Tr-J/+ nerve fibers. The immunocytochemical analysis indicated that PMP-22 protein is associated with type IV collagen (a basal lamina ubiquitous component) in the Tr-J/+ Schwann cell perinuclear region. Changes in mechanical properties of single myelinating Tr-J/+ nerve fibers were investigated, and alterations in cellular stiffness were found. These results might be associated with F-actin cytoskeleton organization in Tr-J/+ nerve fibers. AFM nanoscale imaging focused on topography and mechanical properties of peripheral nerve fibers might provide new insights into the study of peripheral nervous system diseases.


Intravitreal controlled release of dexamethasone from engineered microparticles of porous silicon dioxide.
Dexamethasone is a glucocorticoid that is widely used in the ophthalmic arena. The recent FDA approved dexamethasone implant can provide a three month efficacy but with high rate of drug related cataract and high intraocular pressure (IOP). It seems that higher steroid in aqueous humor and around lens may be associated with these complications based on clinical fact that higher IOP was observed with intravitreal triamcinolone acetonide (TA) than with subtenon TA. We hypothesize that placing a sustained dexamethasone release system near back of the eye through a fine needle can maximize efficacy while mitigate higher rate of IOP rise and cataract. To develop a sustained intravitreal dexamethasone delivery system, porous silicon dioxide (pSiO2) microparticles were fabricated and functionalized with amines as well as carboxyl groups. Dexamethasone was conjugated to pSiO2 through the Steglich Esterification Reaction between hydroxyl of dexamethasone and carboxyl groups on the pSiO2. The drug loading was confirmed by Fourier transform infrared spectroscopy (FTIR) and loading efficiency was quantitated using thermogravimetric analysis (TGA). In vitro release was conducted for three months and dexamethasone was confirmed in the released samples using liquid chromatography-tandem mass spectrometry (LC/MS/MS). A pilot ocular safety and determination of vitreous drug level was performed in rabbit eyes. The drug loading study demonstrated that loading efficiency was from 5.96% to 10.77% depending on the loading reaction time, being higher with longer loading reaction time before reaching saturation around 7 days. In vitro drug release study revealed that dexamethasone release from pSiO2 particles was sustainable for over 90 days and was 80 days longer than free dexamethasone or infiltration-loaded pSiO2 particle formulation in the same setting. Pilot in vivo study demonstrated no sign of ocular adverse reaction in rabbit eyes following a single 3 mg intravitreal injection and free drug level at 2-week was 107.23 ± 10.54 ng/mL that is well above the therapeutic level but only around 20% level of dexamethasone released from OZURDEX(®) (dexamethasone intravitreal implant) in a rabbit eye model. In conclusion, dexamethasone is able to covalently load to the pSiO2 particles and provide sustained drug release for at least 3 months in vitro. Intravitreal injection of these particles were well tolerated in rabbit eyes and free drug level in vitreous at 2-week was well above the therapeutic level.


Development and testing of a measure of health-related quality of life for men with urinary incontinence.
To describe the development and psychometric testing of male versions of the Urogenital Distress Inventory (UDI) and Incontinence Impact Questionnaire (IIQ). Instrument development. Urology clinic at a large urban Veterans Affairs Medical Center. Convenience sample of English-speaking community-dwelling male urology clinic patients who acknowledged concerns about urine leakage. Content experts and male clinic patients evaluated items for clarity and fit. Revised instruments were administered to 153 participants. Data were analyzed to examine issues of feasibility of administration, internal consistency reliability, and validity. Modal completion time for the Male Urogenital Distress Inventory (MUDI) (27 items) and Male Urinary Symptom Impact Questionnaire (MUSIQ) (32 items) was 20 minutes. Cronbach's coefficients were.89 for the MUDI and.95 for the MUSIQ. Total MUDI and MUSIQ scores were moderately correlated (r =.59, P <.001). Mean MUDI and MUSIQ scores varied significantly with self-reported desire for socialization, urine leakage, and depression. Principal components analyses suggested the presence of seven factors accounting for 65.4% of the variance in the MUDI and six factors accounting for 72.9% of the variance in the MUSIQ. It is feasible to use the MUDI and MUSIQ to measure health-related quality of life in men with continence problems. The scores appeared to be reliable and valid in this racially and educationally diverse sample. Use of the MUDI and MUSIQ may provide more sensitive measurement of the specific effect of urinary incontinence and related symptoms on health-related quality of life in men. Future research should determine reproducibility and responsivity and reexamine the construct validity of these instruments.


Unraveling the Complex Olefin Isomer Mixture Using Two-Dimensional Gas Chromatography-Photoionization-Time of Flight Mass Spectrometry.
Commercial dodecenes are a complex chemical mixture with a majority of C12 olefins and minority of C8-18 olefins. Structurally, dodecene products may consist of straight-chain alkenes, branched alkenes, as well as cyclic hydrocarbons. Due to the difference of feeds and catalysts used in the oligomerization reaction, the composition of the dodecenes is complex and their properties are very different. Knowing the complex composition of dodecenes can help tune the production process and select the appropriate products according to their end use. To reveal the complex profile of dodecenes, an analytical method using two-dimensional gas chromatography (GC×GC) coupled photoionization (PI) - time of flight mass spectrometry (TOFMS) was developed in this study. A conventional (nonpolar × polar) column combination (non-polar column as 1st dimension and mid-polar column as 2nd dimension) was selected. The analytical condition of GC was optimized using fractional factorial experimental design (DoE). Olefin congener grouping by carbon chain length and double bond equivalent (DBE) was achieved based on the detection of molecular ions by PI-TOFMS. Grouping of dodecenes by linear, mono-branched, di- and tri-branched subgroups was achieved based on branching index (BI) under the assumption of no retention time (RT) overlap among subgroups. Certain dodecene isomers were identified by retention index (RI) and further confirmed by PI mass spectra. The information altogether provided a multimodal characterization possibility to be used with statistical tools. Principal component analysis (PCA) and hierarchical clustering analysis (HCA) of seventeen dodecene samples explained the composition variance between catalysts solid phosphoric acid and zeolite, as well as between feeds with C4 and without C4.


Antimutagenic effect of garlic (Allium sativum L.) on 4NQO-induced mutagenesis in Escherichia coli WP2.
Aqueous extract prepared from garlic bulbs markedly suppressed the mutagenesis in both E. coli WP2 trp- and E. coli WP2 trp- uvrA- induced by 4-nitroquinoline 1-oxide (4NQO), but not that induced by UV. Cellular toxicity, inhibition of the expression of the Trp+ phenotype and delay of the first cell division after 4NQO treatment were not observed in the presence of the extract. Since the extract showed identical antimutagenic effects against 4NQO in both test strains but no effect on the mutagenesis of UV, it seems that the extract might act by inactivating the electrophilic group(s) of 4NQO or inhibiting its metabolic activation.


Predator-prey interactions and venom composition in a high elevation lizard specialist, Crotalus pricei (Twin-spotted Rattlesnake).
The Twin-spotted Rattlesnake (Crotalus pricei) is a small lizard specialist restricted to higher montane habitat in the Sky Islands of Arizona and México. Though this species is restricted to high elevations and dispersal between mountaintops is impossible, few studies have investigated venom composition or the predator-prey relationship between C. p. pricei and its primary prey source, Yarrow's Spiny Lizard (Sceloporus jarrovii). Because of current isolation of populations in disjunct mountain ranges, it is possible that populations show local adaptation to prey and/or environmental conditions, as reflected by distinct venom phenotypes. Here we characterize venom composition of C. pricei from several different Sky Island mountain ranges and its relationship with S. jarrovii by using various analytical techniques and comparative toxicity tests. Results of venom analyses indicate that there is limited geographic variation in venom composition, occurring primarily in venoms of C. pricei from Durango, México, which had unusual PLA2 isoforms that are lacking from venoms of U.S. populations. Toxicity assays reveal that S. jarrovii has not developed resistance specific to C. p. pricei venom but does display a general tolerance to venom of several snakes in the genus Crotalus. These results provide insight into the evolutionary relationship between a lizard specialist and its natural prey, in addition to novel information on the venom composition of a little-studied species with a narrow range in the United States.


Seroprevalence of Toxoplasma gondii infection among immigrant and native pregnant women in Eastern Spain.
In European countries, toxoplasma antenatal screening is recommended to prevent toxoplasmosis. The seroprevalence of these infections in immigrants can be different than in native population. From February 2006 to June 2010, a cross-sectional study was carried out in all pregnant women attended at a reference unit in Elche, Spain. An enzyme immunoassay was used for detection of IgG antibodies against Toxoplasma gondii. For each immigrant woman, one Spanish pregnant woman of the same age cared for in the same day was recruited (Spanish control group). A total of 1,627 migrant pregnant women participated in this study. The adherence to screening among migrants was 91.9% (95% CI, 90.5-93.1%), similar than that found in Spaniards (92.2%; 95% CI, 90.8-93-4%). Among migrant women, 619 were positive for IgG anti-T. gondii antibodies (41.4%; 95% CI, 38.9-43.9%), compared with 12.0% (95% CI, 10.5-13.8%) among Spaniards (odds ratio (OR), 5.2 (95% CI, 4.3-6.3). Seroprevalence in pregnant women from Latin America, northern Africa, Eastern Europe, Africa Sub-Saharan and Western Europe was higher than in the Spanish control group (OR, 5.4, 5.8, 6.5, 5.4, and 2.4, respectively; p < 0.001). No Asian pregnant woman was immune. Seroprevalence increased with increasing age in migrant pregnant women: 15-25 years, 38.2%; 26-35 years, 40.7%; and 36-45 years, 52.8%. The seroprevalence of T. gondii infection in migrant pregnant women living in Spain was higher than in the native population. However, no cases were found in Asian immigrants, highlighting the importance of primary prevention of this infection in pregnant women coming from that geographic region.


Wilms' tumor 1 enhances Cisplatin-resistance of advanced NSCLC.
Wilms' tumor 1 (WT1) is an oncogene that has been correlated with tumor progression, bad prognosis and chemo-resistance in Non-Small-Cell lung cancer (NSCLC). Here, we found that WT1 expression is significantly higher in NSCLCs than in benign controls, and that Cisplatin-resistant patients display a notable increase in WT1 expression following relapse. In vitro, WT1 levels were associated with the IC50 of NSCLC cells and increased along with treatment time and dose of Cisplatin. Furthermore, WT1 enhanced Major Vault Protein (MVP) transcription via binding to its promoter. Therefore, WT1 may be a potential therapeutic target for solving resistance.


Prenatal choline supplementation during mouse pregnancy has differential effects in alcohol-exposed fetal organs.
Fetal alcohol spectrum disorders (FASD) are preventable adverse outcomes consequent to prenatal alcohol exposure. Supplemental choline confers neuroprotection to the alcohol-exposed offspring, but its actions outside the brain are unclear. We previously reported that prenatal exposure of mice to 4.5 g/kg of alcohol decreased placental weight in females only, but decreased body weight and liver-to-body weight ratio and increased brain-to-body weight ratio in both sexes. Here we test the hypotheses that a lower alcohol dose will elicit similar outcomes, and that concurrent choline treatment will mitigate these outcomes. Pregnant C57BL/6J mice were gavaged with alcohol (3 g/kg; Alc) or maltodextrin (MD) from embryonic day (E) 8.5-17.5. Some also received a subcutaneous injection of 100 mg/kg choline chloride (Alc + Cho, MD + Cho). Outcomes were evaluated on E17.5. Alc dams had lower gestational weight gain than MD; this was normalized by choline. In males, Alc decreased placental weight whereas choline increased placental efficiency, and Alc + Cho (vs. MD) tended to further reduce placental weight and increase efficiency. Despite no significant alcohol effects on these measures, choline increased fetal body weight but not brain weight, thus reducing brain-to-body weight ratio in both sexes. This ratio was also lower in the Alc + Cho (vs. MD) fetuses. Alc reduced liver weight and the liver-to-body weight ratio; choline did not improve these. Placental weight and efficiency correlated with litter size, whereas placental efficiency correlated with fetal morphometric measurements. Choline prevents an alcohol-induced reduction in gestational weight gain and fetal body weight and corrects fetal brain sparing, consistent with clinical findings of improvements in alcohol-exposed children born to mothers receiving choline supplementation. Importantly, we show that choline enhances placental efficiency in the alcohol-exposed offspring but does not normalize fetal liver growth. Our findings support choline supplementation during pregnancy to mitigate the severity of FASD and emphasize the need to examine choline's actions in different organ systems.


Non-ulcer dyspepsia in the long-term perspective.
To study the natural history of non-ulcer dyspepsia. Retrospective study on consecutive cases of non-ulcer dyspepsia diagnosed in 1980 and 1981. The long-term course was ascertained from medical records and from answers to a questionnaire completed by the patients at 10-year follow-up. Out-patients at the endoscopy unit of a county hospital. The diagnosis was based on typical symptoms of dyspepsia, a normal index endoscopy and a lack of evidence of organic disease related to the symptoms during the subsequent 12-month period. Two hundred and seventy-one patients were entered into the study. Of the 195 who received a questionnaire at 10-year follow-up, 165 (85%) responded. The median age at inclusion was 51 years (25-75 percentiles 38-65 years), and 130 men and 141 women were included in the study. The cumulative death rate after 10 years was 18%, a value no greater than that in the general population: 8% had died of cardiovascular disease, 1.5% of gastrointestinal cancer and 3% of other malignancies. One patient had died of a perforated gastric ulcer. At 10-year follow-up, a further three patients had developed peptic ulcers (expected number 4.2), 64% had experienced dyspeptic symptoms during the preceding 12 months and 41% had been taking analgesics during the previous week. Of the symptoms listed by the patients, 30% were typical of irritable bowel syndrome. Non-organic dyspepsia seems to be a chronic disease in most patients. The course is benign and unrelated to peptic ulcer disease.


Di-μ(2)-methano-lato-bis-(μ-4-methyl-5-sulfanyl-idene-4,5-dihydro-1H-1,2,4-triazolido-κN:N)di-μ(3)-oxido-tetra-kis-[dimethyl-tin(IV)].
The title distannoxane, [Sn(4)(CH(3))(8)(C(3)H(4)N(3)S)(2)(CH(3)O)(2)O(2)], lies about a center of inversion; the tetra-nuclear mol-ecule features a three-rung-staircase Sn(4)O(4) core in which the two crystallographically independent Sn(IV) atoms are bridged by the triazolide group. The negatively charged N atom of the triazolide group binds to the terminal Sn atom at a shorter distance [Sn-N = 2.239 (2) Å] compared with the neutral N atom that binds to the central Sn atom [Sn← N = 2.757 (3) Å]. The oxide O atom is three-coordinate whereas the methano-late O atom is two-coordinate. The terminal Sn atom is five-coordinate in a cis-C(3)SnNO trigonal-bipyramidal environment, whereas the central Sn atom is six-coordinate in a C(2)SnNO(3) skew-trapezoidal-bipyramidal geometry.


Effect of high pressure and thermal processing on shelf life and quality of strawberry purée and juice.
The purpose of the study was to determine the effects of high pressure processing (HPP; 400-600 MPa, 20 °C, 1.5 or 3 min) and heat treatment (HT; 85 °C for 2 min) of strawberry purée and juice made from the same raw material. Microbiological and enzymatic inactivation, Brix, pH, anthocyanins, vitamin C, colour and sensory properties were analysed after processing and cold storage. The microbiological shelf life of the products was at least 49 days when processed at 500 or 600 MPa. Anthocyanins, vitamin C and colour were well preserved after HPP and HT. During storage, anthocyanins, vitamin C and sensory quality were better conserved in HT than in HPP purées, while there were minor differences between HT and HPP juices. This was probably due to higher enzyme activity in HPP purées, and indicates that raw materials with lower initial enzyme activity, like juices, are more suited for HPP than e.g. purées.