Indo-Pacific Walker circulation drove Pleistocene African aridification.
Today, the eastern African hydroclimate is tightly linked to fluctuations in the zonal atmospheric Walker circulation1,2. A growing body of evidence indicates that this circulation shaped hydroclimatic conditions in the Indian Ocean region also on much longer, glacial-interglacial timescales3-5, following the development of Pacific Walker circulation around 2.2-2.0 million years ago (Ma)6,7. However, continuous long-term records to determine the timing and mechanisms of Pacific-influenced climate transitions in the Indian Ocean have been unavailable. Here we present a seven-million-year-long record of wind-driven circulation of the tropical Indian Ocean, as recorded in Mozambique Channel Throughflow (MCT) flow-speed variations. We show that the MCT flow speed was relatively weak and steady until 2.1 ± 0.1 Ma, when it began to increase, coincident with the intensification of the Pacific Walker circulation6,7. Strong increases during glacial periods, which reached maxima after the Mid-Pleistocene Transition (0.9-0.64 Ma; ref. 8), were punctuated by weak flow speeds during interglacial periods. We provide a mechanism explaining that increasing MCT flow speeds reflect synchronous development of the Indo-Pacific Walker cells that promote aridification in Africa. Our results suggest that after about 2.1 Ma, the increasing aridification is punctuated by pronounced humid interglacial periods. This record will facilitate testing of hypotheses of climate-environmental drivers for hominin evolution and dispersal.


Correlation between kidney transplantation and colorectal cancer in hemodialysis patients: A nationwide, retrospective, population-based cohort study.
Kidney transplantation (KT) correlates with an increased risk of developing several malignancies; however, the risk of colorectal cancer (CRC) after KT remains debatable and has been marginally explored. Hence, in this nationwide, retrospective, population-based cohort study, we aimed to examine the correlation between KT and CRC in a large-scale population-based Chinese cohort. We identified a total of 3739 regular hemodialysis patients undergoing KT (exposed cohort) and 42,324 hemodialysis patients not undergoing KT (non-exposed cohort) between 2000 and 2008 from Taiwan's National Health Insurance Research Database (NHIRD). Both cohorts were followed up from January 1, 2000, to the date of CRC diagnosis, death, or the end of 2013. Using Kaplan-Meier method, we measured the cumulative incidence of CRC in each cohort. Furthermore, Cox proportional hazards models were used to compute hazards ratios (HRs) and 95% confidence intervals (CIs) to estimate the correlation between KT and CRC in hemodialysis patients. The Kaplan-Meier analysis revealed that the cumulative incidence of CRC was significantly higher in the exposed cohort than in the non-exposed cohort (log-rank test, P < 0.001). After adjusting for potential confounders, the exposed cohort exhibited a significantly increased risk of CRC compared with the non-exposed cohort (adjusted HR, 1.34; 95% CI, 1.11-1.62). Hemodialysis patients undergoing KT have a significantly higher risk of CRC than those not undergoing KT. Cancer should continue to be a primary focus of prevention during KT.


Non-cardia gastric adenocarcinoma more than 5 years after laparoscopic sleeve gastrectomy: A case report and literature review.
Obesity is associated with an increased risk of several cancers including adenocarcinomas of the esophagus and gastric cardia but not with distal gastric cancer (GC). Laparoscopic sleeve gastrectomy (LSG) is a popular bariatric procedure. We report the fifth case in the medical literature of non-cardia gastric adenocarcinoma 63 months after LSG in a 26-year-old woman with no known recognizable risk factors. Among these five cases, the median (range) age at time of diagnosis of GC was 47 (26-64) years, the body mass index (BMI) prior to LSG was 47 (35.3-65.1) kg/m2 , the BMI at time of cancer diagnosis was 32 (29.3-47.0) kg/m2 , and the interval between LSG and diagnosis was 54 (9-63) months. Nonetheless, the link between LSG and the risk of GC remains skeptical and is discussed.


Social and geographic inequalities in premature adult mortality in Japan: a multilevel observational study from 1970 to 2005.
To examine trends in social and geographic inequalities in all-cause premature adult mortality in Japan. Observational study of the vital statistics and the census data. Japan. Entire population aged 25 years or older and less than 65 years in 1970, 1975, 1980, 1985, 1990, 1995, 2000 and 2005. The total number of decedents was 984 022 and 532 223 in men and women, respectively. For each sex, ORs and 95% CIs for mortality were estimated by using multilevel logistic regression models with 'cells' (cross-tabulated by age and occupation) at level 1, 8 years at level 2 and 47 prefectures at level 3. The prefecture-level variance was used as an estimate of geographic inequalities of mortality. Adjusting for age and time-trends, compared with production process and related workers, ORs ranged from 0.97 (95% CI 0.96 to 0.98) among administrative and managerial workers to 2.22 (95% CI 2.19 to 2.24) among service workers in men. By contrast, in women, the lowest odds for mortality was observed among production process and related workers (reference), while the highest OR was 12.22 (95% CI 11.40 to 13.10) among security workers. The degree of occupational inequality increased in both sexes. Higher occupational groups did not experience reductions in mortality throughout the period and was overtaken by lower occupational groups in the early 1990s, among men. Conditional on individual age and occupation, overall geographic inequalities of mortality were relatively small in both sexes; the ORs ranged from 0.87 (Okinawa) to 1.13 (Aomori) for men and from 0.84 (Kanagawa) to 1.11 (Kagoshima) for women, even though there is a suggestion of increasing inequalities across prefectures since 1995 in both sexes. The present findings suggest that both social and geographic inequalities in all-cause mortality have increased in Japan during the last 3 decades.


Distinct dynamics of social motivation drive differential social behavior in laboratory rat and mouse strains.
Mice and rats are widely used to explore mechanisms of mammalian social behavior in health and disease, raising the question whether they actually differ in their social behavior. Here we address this question by directly comparing social investigation behavior between two mouse and rat strains used most frequently for behavioral studies and as models of neuropathological conditions: C57BL/6 J mice and Sprague Dawley (SD) rats. Employing novel experimental systems for behavioral analysis of both subjects and stimuli during the social preference test, we reveal marked differences in behavioral dynamics between the strains, suggesting stronger and faster induction of social motivation in SD rats. These different behavioral patterns, which correlate with distinctive c-Fos expression in social motivation-related brain areas, are modified by competition with non-social rewarding stimuli, in a strain-specific manner. Thus, these two strains differ in their social behavior, which should be taken into consideration when selecting an appropriate model organism.


Streamlined Purification of RNA-Protein Complexes Using UV Cross-Linking and RNA Antisense Purification.
RNA-protein interactions are important in development and disease, but identification of novel RNA-protein interactions remains challenging. Here, we describe an updated capture method to identify direct and specific RNA-protein interactions. First, RNA and protein are covalently cross-linked in living cells by treatment with UV light at 254 nanometers wavelength. The antisense purification approach is dependent upon nucleic acid hybridization between biotinylated DNA probes and a target RNA. Target protein:RNA:DNA complexes are enriched by capture on streptavidin magnetic beads and purified through several denaturing washes that remove nonspecific protein and nucleic acid interactors. Mass spectrometry is used to identify proteins that are specifically enriched in the target RNA capture. This method has been applied to discover the protein interactions of noncoding RNAs but can be used to capture any RNA where the target sequence is known.


Decrease in complications during cataract surgery with the use of a silicone-tipped irrigation/aspiration instrument.
To compare rates of vitreous loss during cataract surgery when a silicone-tipped irrigation/aspiration (I/A) instrument is used for cortex removal as opposed to a metal tip. Parkland Memorial Hospital, Dallas, Texas, USA. A retrospective chart review of all patients who had cataract extraction by phacoemulsification by third-year ophthalmology residents between September 2000 and February 2004 was conducted. A silicone-tipped I/A instrument was used to remove cortex for all surgeries beginning in September 2002, whereas a metal I/A tip was used previously. Of the 1072 cases performed with a metal I/A tip, there were 13 cases of vitreous loss during cortex removal (rate 1.2%) and 26% of all vitreous loss during that time occurred during cortex removal. Of the 805 cases performed with a silicone I/A tip, there was a single case (0.1%) of vitreous loss during cortex removal (P=.004); only 4% of all vitreous loss during that time occurred during cortex removal (P=.011). The overall incidence of vitreous loss during cortex removal and the ratio of vitreous loss during cortex removal to all cases of vitreous loss were significantly decreased using the silicone-tipped I/A instrument.


Solubility of pharmaceutical ingredients in triglycerides.
Lipid-based drug delivery systems (LBDDS) are highly relevant as pharmaceutical formulations significantly enhancing the bioavailability of active pharmaceutical ingredients (APIs). These formulations often are complex mixtures of APIs, various lipids, and other excipients (e.g. surfactants). In their simplest form, LBDDS contain one API being dissolved in a pure lipid, which often is a triglyceride (TG). In this work, solubilities of the APIs indomethacin, ibuprofen, and fenofibrate in pure TGs of different chain lengths (C chain 8-18) and degree of saturation were investigated. Solubilities of APIs in TGs were measured via differential scanning calorimetry, hot-stage microscopy, high-performance liquid chromatography, and Raman spectroscopy. The influence of fatty-acid chain length and degree of saturation on the API solubility in the TGs was investigated. APIs showed a higher solubility in saturated (wIBU = 10.5 wt% at 25 °C in tricaprylin) TGs compared to unsaturated ones (wIBU = 4.0 wt% at 25 °C in triolein). The fatty-acid chain length of TGs only slightly affects the solubility of ibuprofen and fenofibrate, but strongly influences the eutectic temperature of the API/TG mixtures. API solubilities in TGs and TG mixtures (mixtures of tricaprylin and tricaprin) were successfully modeled using the Perturbed-Chain Statistical Associating Fluid Theory (PC-SAFT) accounting for the intermolecular API/TG interactions providing a deep understanding of the energetic and structural impact of the TGs on API solubility.


Influence of preoperative showers on staphylococcal skin colonization: a comparative trial of antiseptic skin cleansers.
We undertook a prospective randomized observer-blinded study comparing the ability of preoperative showers with chlorhexidine gluconate (Hibiclens), povidone-iodine (Betadine), and a lotion soap (Safe 'N Sure) to diminish the staphylococcal skin flora of patients. By block randomization, patients scheduled for an elective cardiac operation or coronary artery angioplasty were assigned to shower with one of the study skin cleansers either once (evening only) or twice (both evening and morning) before the procedure. Semiquantitative samples for culture were obtained from the subclavian and inguinal sites on the evening before the procedure (baseline culture) and again the next morning before the operation. The chlorhexidine skin cleanser consistently reduced staphylococcal colony counts at both the subclavian and inguinal sites before the procedure. This reduction was significant for patients showering both evening and morning (p less than 0.05). The use of the povidone-iodine skin cleanser inconsistently affected skin flora. Patients using lotion soap either experienced no change or had an increase in colony counts. Chlorhexidine is more effective than povidone-iodine in diminishing skin colonization with staphylococci in patients before operation. Repeated applications of chlorhexidine are superior to a single shower with this agent.


Endoplasmic reticulum targeted chemotherapeutics: the remarkable photo-cytotoxicity of an oxovanadium(IV) vitamin-B6 complex in visible light.
An oxovanadium(IV) vitamin-B6 Schiff base complex, viz. [VO(HL)(acdppz)]Cl, having (acridinyl)dipyridophenazine (acdppz) shows specific localization to endoplasmic reticulum (ER) and remarkable apoptotic photocytotoxicity in visible light (400-700 nm) in HeLa and MCF-7 cancer cells (IC50 < 0.6 μM) while being non-toxic in the dark and to MCF-10A normal cells (IC50 > 40 μM).


Fabrication and Characterization of Poly(lactic acid)/Poly(ε-caprolactone) Blend Electrospun Fibers Loaded with Amoxicillin for Tunable Delivering.
Blends of poly(lactic acid) (PLA) and poly(ε-caprolactone) (PCL), loaded with different amounts of Amoxicillin antibiotic (AMOX) were electrospun to investigate their release properties and obtain a controlled and tuneable release. The processing parameters for electrospinning were set up and reliable membranes were obtained. Morphology and thermal behaviour were found dependent on the component ratio as well as on the incorporated drug amount. A very different release kinetics of the two pristine polymers, very rapid for PCL and very slow for PLA, reflected in intermediate release time. However comparing the release amount with that predicted by the mixture rule a preferential incorporation of AMOX into PLA can be inferred.


The mitochondrial protein TIMM44 is required for angiogenesis in vitro and in vivo.
The mitochondrial integrity and function in endothelial cells are essential for angiogenesis. TIMM44 (translocase of inner mitochondrial membrane 44) is essential for integrity and function of mitochondria. Here we explored the potential function and the possible mechanisms of TIMM44 in angiogenesis. In HUVECs, human retinal microvascular endothelial cells and hCMEC/D3 brain endothelial cells, silence of TIMM44 by targeted shRNA largely inhibited cell proliferation, migration and in vitro capillary tube formation. TIMM44 silencing disrupted mitochondrial functions in endothelial cells, causing mitochondrial protein input arrest, ATP reduction, ROS production, and mitochondrial depolarization, and leading to apoptosis activation. TIMM44 knockout, by Cas9-sgRNA strategy, also disrupted mitochondrial functions and inhibited endothelial cell proliferation, migration and in vitro capillary tube formation. Moreover, treatment with MB-10 ("MitoBloCK-10"), a TIMM44 blocker, similarly induced mitochondrial dysfunction and suppressed angiogenic activity in endothelial cells. Contrarily, ectopic overexpression of TIMM44 increased ATP contents and augmented endothelial cell proliferation, migration and in vitro capillary tube formation. In adult mouse retinas, endothelial knockdown of TIMM44, by intravitreous injection of endothelial specific TIMM44 shRNA adenovirus, inhibited retinal angiogenesis, causing vascular leakage, acellular capillary growth, and retinal ganglion cells degeneration. Significant oxidative stress was detected in TIMM44-silenced retinal tissues. Moreover, intravitreous injection of MB-10 similarly induced oxidative injury and inhibited retinal angiogenesis in vivo. Together, the mitochondrial protein TIMM44 is important for angiogenesis in vitro and in vivo, representing as a novel and promising therapeutic target of diseases with abnormal angiogenesis.


Ciliary neurotrophic factor-mediated neuroprotection involves enhanced glycolysis and anabolism in degenerating mouse retinas.
Ciliary neurotrophic factor (CNTF) acts as a potent neuroprotective cytokine in multiple models of retinal degeneration. To understand mechanisms underlying its broad neuroprotective effects, we have investigated the influence of CNTF on metabolism in a mouse model of photoreceptor degeneration. CNTF treatment improves the morphology of photoreceptor mitochondria, but also leads to reduced oxygen consumption and suppressed respiratory chain activities. Molecular analyses show elevated glycolytic pathway gene transcripts and active enzymes. Metabolomics analyses detect significantly higher levels of ATP and the energy currency phosphocreatine, elevated glycolytic pathway metabolites, increased TCA cycle metabolites, lipid biosynthetic pathway intermediates, nucleotides, and amino acids. Moreover, CNTF treatment restores the key antioxidant glutathione to the wild type level. Therefore, CNTF significantly impacts the metabolic status of degenerating retinas by promoting aerobic glycolysis and augmenting anabolic activities. These findings reveal cellular mechanisms underlying enhanced neuronal viability and suggest potential therapies for treating retinal degeneration.


The perception of temporal deviations in isochronic patterns.
In a study of perceptual synchronization with an isochronic sequence, subjects were given the following task: They heard an isochronic sequence of tones in which the last interval was either correct or too long. Their task was to detect irregularity. The independent variables were the number of tones heard and the time interval between them. The dependent variable was the difference limen (DL) for the detectability of the irregularity. Two experiments were performed in this study, differing in the way in which the trials were blocked: In Experiment 1, stimuli with the same period were presented in blocks, whereas in Experiment 2, the period of the stimulus was randomized. The results show that in Experiment 1 the number of tones in the stimulus did not affect the detectability of the anisochrony. In Experiment 2, the number of the DL was a decreasing function of the number of tones heard. Moreover, the decrease of the DL was larger than one would expect from a simple model of information integration, which assumes that subjects improve their performance by averaging their percepts of the first intervals in the sequence. The difference between this task and experiments on the discrimination of temporal intervals is discussed.


Sodium excretion after bilateral adrenalectomy in rats with experimental cirrhosis.
Bilateral adrenalectomy (ADX) or a sham procedure was performed in cirrhotic and control rats. After ADX, controls increased their sodium excretion with respect to the basal values but cirrhotics did not. When sodium-loaded, the ADX cirrhotic rats retained a significant amount of the load. These data do not support a primary role of aldosterone in the impaired sodium handling by cirrhotic rats.


Integrated, DC voltage-driven nucleic acid diagnostic platform for real sample analysis: Detection of oral cancer.
We present an integrated and low-cost microfluidic platform capable of extraction of nucleic acids from real biological samples. We demonstrate the application of this platform in pathogen detection and cancer screening. The integrated platform consists of three units including a pretreatment unit for separation of nucleic acids from lysates, a preconcentration unit for concentration of isolated nucleic acids and a sensing unit localized at a designated position on the chip for specific detection of the target nucleic acid. The platform is based on various electrokinetic phenomena exhibited by ion exchange membranes in a DC electrical field that allow them to serve as molecular filters, analyte preconcentrators and sensors. In this manuscript, we describe each unit of the integrated chip separately and show specific detection of a microRNA (miRNA 146a) biomarker associated with oral cancer as a proof-of-concept experiment. This platform technology can easily be extended to other targets of interest by optimizing the properties of the ion exchange membranes and the specific probes functionalized onto the sensors.


Induction of Ia antigens on thyroid cultures by syngeneic T lymphocytes: a model of autoimmune disease.
We previously reported that the expression of Ia antigens on cultured monolayers of murine thyroid epithelial cells (TEC) occurred with a particular distribution exclusively on the basal part of the cultured thyroid cells, while class I antigens of the major histocompatibility complex (MHC) are only detected on the apical surface. It appears that deposition of syngeneic lymphocytes induces, 24 h later, Ia expression on the apical side of cultured TEC, the surface that is in direct contact with the responder lymphocytes during syngeneic sensitization of T lymphocytes. We hypothesized that this phenomenon could represent, in syngeneic situations, the restriction process in antigen recognition by T cells, as demonstrated by Ia restricted primary syngeneic sensitization (PSS) on murine TEC.


Osteosynthesis of mallet finger using plate and screws: evaluation of 25 patients.
To evaluate the results from surgical treatment of patients with mallet finger injury using a hook plate and screw. Twenty-five patients (19 males and six females) between the ages of 20 and 35 years were analyzed between May 2008 and December 2012. They were evaluated in accordance with Crawford's criteria and the mean follow-up was 18 months. The results from 10 patients (40%) were excellent and from 15 (60%), good. Twenty-one patients (84%) reported no pain, 18 months after the operation. There was no limitation to range of motion in 14 cases (56%), limitation of extension in seven (28%) and limitation of flexion in four (16%). Surgical treatment by means of open reduction and internal fixation using a hook plate and screw proved to be an excellent option for treating mallet finger fractures and was considered to be a safe and effective method.


Religiousness and religious coping in a secular society: the gender perspective.
Women are found to be more religious than men and more likely to use religious coping. Only few studies have explored religious gender differences in more secular societies. This population-based study comprised 3,000 Danish men and women (response rate 45 %) between 20 and 40 years of age. Information about demographics, religiousness and religious coping was obtained through a web-based questionnaire. We organized religiousness in the three dimensions: Cognition, Practice and Importance, and we assessed religious coping using the brief RCOPE questionnaire. We found substantial gender differences in both religiousness and religious coping. Nearly, 60 % of the women believed in some sort of spirit or in God compared to 40 % of the men. Generally, both men and women scored low on the RCOPE scale. However, for respondents reporting high levels of religiousness, the proportion of men who scored high in the RCOPE exceeded the proportion of women in using positive and especially negative coping strategies. Also, in a secular society, women are found to be more religious than men, but in a subset of the most religious respondents, men were more inclined to use religious coping. Further studies on religious coping in secular societies are required.


The 2019 ESPEN Arvid Wretlind lecture perioperative nutritional and metabolic care: Patient-tailored or organ-specific approach?
The perioperative severe changes in the nutritional and metabolic homeostasis are, by some means, proportional to the extent of tissue injury and magnitude of operative trauma. An adequate qualitative and quantitative replacement of nutritional substrates are of utmost importance to facilitate proper tissue healing and recovery and maintenance of organ function after surgery. The present manuscript has been planned to put the most recent research of the Milano-Bicocca University surgical working group in the context of a more personalized nutritional therapy and metabolic care for surgical patients. Particular prominence has been given to major pancreatic resections because these surgeries are among the most complex and challenging operations for the degree of parenchyma resection and tissue dissection, the consequent overall injury, and the fairly high rate of major complications resulting in a catabolic response. Anthropometric parameters and particularly sarcopenia, visceral obesity - and their relative proportion -, are strongly associated with poor outcome after pancreatic surgery. Adequate perioperative nutritional therapy is of utmost importance in affecting morbidity. Long-term nutritional and metabolic sequelae, caused by exocrine pancreatic insufficiency, need to be promptly recognized and treated with an adequate enzyme supplementation. There is strong evidence sustaining the necessity of proper perioperative metabolic and nutritional care into the management of patients undergoing major pancreatic surgery.


Safety and tolerability of once-daily controlled-release carvedilol 10-80 mg in Japanese patients with chronic heart failure.
The aim of the present study was to assess the safety and tolerability of the controlled-release (CR) formulation of the β-blocker carvedilol in Japanese patients with chronic heart failure (HF). In this multicenter, randomized, open-label, phase I/II dose-escalation study, 41 patients receiving standard therapy for chronic HF were randomized in a ratio of 1:1 to carvedilol CR or immediate-release (IR) carvedilol. The primary objective was to evaluate the tolerability and safety of escalating doses of carvedilol CR (10-40 mg/day), with a reference arm of 5-20 mg/day of carvedilol IR. In addition, the tolerability and safety of titration to a carvedilol CR dose up to 80 mg/day were examined, as were plasma concentrations of carvedilol and changes in vital signs. The proportions of patients who completed 40-mg/day carvedilol CR and 20-mg/day carvedilol IR were 42% (8/19) and 50% (11/22), respectively. In the CR group, 7/19 (37%) attained a dose of 80 mg. During the primary evaluation period, 7/19 (37%) and 4/22 (18%) patients experienced drug-related adverse events in the CR and IR groups, respectively, the characteristics of which were similar between groups. No new safety issues emerged in Japanese chronic HF patients treated with carvedilol CR in contrast to those known in carvedilol IR.


Purification, characterization and bactericidal activities of basic phospholipase A2 from the venom of Agkistrodon halys (Chinese pallas).
Agkistrodon snake venoms contain a variety of phospholipases (PLA2), some of which are myotoxic. In this study, we used reverse-phase HPLC to purify PLA2 from the venom of Agkistrodon halys. The enzyme named as AgkTx-II, a basic Asp49 PLA2, has a molecular masses of 13,869.05. The amino acid sequence and molecular mass of AgkTx-II was identical to those of an Asp49 basic myotoxic PLA2 previously isolated from this venom. Antibacterial activities were tested by susceptibility and broth-dilution assays. AgkTx-II exerted a potent antibacterial activity against Staphylococcus aureus, Proteus vulgaris, Proteus mirabilis, and Burkholderia pseudomallei. The MIC values of AgkTx-II ranged between 85 and 2.76microM and was most effective against S. aureus, P. vulgaris, P. mirabilis (MIC of 21.25microM) and B. pseudomallei (MIC of 10.25microM). This AgkTx-II rapidly killed S. aureus, P. vulgaris and B. pseudomallei in a dose-dependent manner. The effect of the AgkTx-II on bacterial membranes was evaluated by scanning and transmission electron microscopy. AgkTx-II caused morphological alterations apparent on their cellular surfaces, suggesting a killing mechanism based on membrane permeabilization and damage. Cytotoxicity was measured by XTT tetrazolium (2,3-bis[2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide) and lactate dehydrogenase (LDH) assays using U-937 cells (monocytes). The AgkTx-II did not affect cell viability up to 500microM concentrations but cell death was evident at 1000microM concentration after 24 and 48h. Furthermore, the repeated exposure of AgkTx-II (2-14microM) treated mice showed different tissue alterations, mainly at the brain and kidney; the toxicological potential of AgkTx-II remains to be elucidated. The AgkTx-II exhibits no hemolytic action even at high doses (10-100microM) in human erythrocytes. However, the AgkTx-II is believed to exert its bactericidal effect by permeabilizing the bacterial membrane by forming pores. In addition, the basic PLA2 AgkTx-II displays a bactericidal effect, which may be either dependent or independent of catalysis.


Viscosin-like lipopeptides from frog skin bacteria inhibit Aspergillus fumigatus and Batrachochytrium dendrobatidis detected by imaging mass spectrometry and molecular networking.
Amphibian populations worldwide have declined and in some cases become extinct due to chytridiomycosis, a pandemic disease caused by the fungus Batrachochytrium dendrobatidis; however, some species have survived these fungal epidemics. Previous studies have suggested that the resistance of these species is due to the presence of cutaneous bacteria producing antifungal metabolites. As our understanding of these metabolites is still limited, we assessed the potential of such compounds against human-relevant fungi such as Aspergillus. In this work we isolated 201 bacterial strains from fifteen samples belonging to seven frog species collected in the highlands of Panama and tested them against Aspergillus fumigatus. Among the 29 bacterial isolates that exhibited antifungal activity, Pseudomonas cichorii showed the greatest inhibition. To visualize the distribution of compounds and identify them in the inhibition zone produced by P. cichorii, we employed MALDI imaging mass spectrometry (MALDI IMS) and MS/MS molecular networking. We identified viscosin and massetolides A, F, G and H in the inhibition zone. Furthermore, viscosin was isolated and evaluated in vitro against A. fumigatus and B. dendrobatidis showing MIC values of 62.50 µg/mL and 31.25 µg/mL, respectively. This is the first report of cyclic depsipeptides with antifungal activity isolated from frog cutaneous bacteria.


A suburethral sling procedure with polytetrafluoroethylene for the treatment of genuine stress incontinence in patients with low urethral closure pressure.
One indication for suburethral sling procedures has been recurrent genuine stress incontinence after previous incontinence surgery. Patients with low urethral closure pressures (20 cm H2O or less) in association with genuine stress incontinence are at particular risk for failure of standard anti-incontinence procedures. Urodynamic evaluation was used to select 17 patients with genuine stress incontinence and low urethral closure pressures for surgical treatment with a sling procedure using polytetrafluoroethylene. The technique of the procedure, cure rate, and postoperative complications were assessed. An 85% subjective and objective cure rate was found on urodynamic testing three months postoperatively. Complications included wound seroma, urinary tract infection, and urinary retention.


Current prospects for mRNA gene delivery.
Replication-deficient viruses have been used most successfully in the field of gene therapy because of their high transfection efficiency. However, the risk of insertional mutagenesis and induction of unwanted immune responses remains still critical for their safe application. On the other hand, nonviral vectors have been intensively investigated for plasmid DNA (pDNA) delivery as a safer alternative although their gene transfer efficiency is still many folds lower than for viral vectors, which has been predominately attributed to the insufficient transport of pDNA into the nucleus. Instead of pDNA, messenger RNA (mRNA) has recently emerged as an attractive and promising alternative in the nonviral gene delivery field. This strategy combines several advantages compared to pDNA: (i) the nuclear membrane, which is a major obstacle for pDNA, can be avoided because mRNA exerts its function in the cytoplasm; (ii) the risk of insertional mutagenesis can be excluded; (iii) the determination and use of an efficient promoter is omitted; (iv) repeated application is possible; (v) mRNA is also effective in non-dividing cells, and (vi) vector-induced immunogenicity may be avoidable. In this review, we summarize recent improvements of mRNA gene delivery and discuss its opportunities for the usage in gene therapy.


[Objective assessment of the tissue trauma in surgery of endometrial cancer].
To review the current possibilities of objective assessment of tissue trauma in surgical treatment of endometrial cancer. Review article. Department of Obstetrics and Gynaecology, Faculty of Medicine and Dentistry, Palacky University in Olomouc. After critical evaluation of relevant contemporary literature, selected methods of the objective estimation of the extent of tissue trauma are presented. The aim of their choice was to find methods objectivly assessing the extent of tissue trauma in connection with using different types of surgical procedures in endometrial cancer treatment. The following clinical and biochemical markers - VAS, KO, CRP, IL-6, neopterin, kynurenin, retinol, α-tocoferol and citrulin seem to be best suitable for the assessment of surgical trauma. These markers should objectivly evaluate the peroperative burden in different types of surgical procedures used for treatment of endometrial cancer.


Auriculotherapy for sleep quality in people with primary insomnia: A protocol for a systematic review and meta-analysis.
Primary insomnia is one of the common sleep disorders. Auriculotherapy originated from traditional Chinese medicine, has been thought as a promising treatment for primary insomnia. We aim to evaluate the efficacy and safety of auriculotherapy for patients with primary insomnia through this systematic review. Five English databases (Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, AMED, and CINAHL), 4 Chinese databases (CBM, CNKI, CQVIP, and Wanfang), and 5 clinical trial registration databases (ClinicalTrials.gov, ANZCTR, EU-CTR, ChiCTR, and ICTRP) will be searched from establishment of the database until November 2018. Articles written in English or Chinese languages will be included. The randomized controlled trials of auriculotherapy (auricular acupuncture and auricular acupressure) for patients with primary insomnia will be included. The primary outcome will be assessed according to the Pittsburgh sleep quality index. Meta-analysis will be conducted with the use of RevMan 5.3. The specific process will refer to the Cochrane Handbook 5.1 for Systematic Review. High-quality synthesis of current evidence on the efficacy and safety of auriculotherapy for primary insomnia will be provided in this study. This systematic review aims to present evidence for whether auriculotherapy is an effective intervention which can improve sleep quality in patients suffering primary insomnia. CRD42019106422.


Right ventricular compression observed in echocardiography from pectus excavatum deformity.
Pectus excavatum exists as varying anatomic deformities and compression of the right heart by the chest wall can lead to patient symptoms including dyspnea and chest pain with exertion. Echocardiography can be difficult but is critical to the evaluation and diagnosis of this patient population. Modifying standard views such as biplane transthoracic and 3-D transesophageal views may be necessary in some patients due to limitations from the abnormal anatomy of the deformed anterior chest wall. Apical four-chamber views when seen clearly can usually visualize any extrinsic compression to the right ventricle of the heart.


Linearization of two ligand-one binding site scatchard plot and the "IC50" competitive inhibition plot: application to the simplified graphical determination of equilibrium constants.
A simple procedure for linearizing the curved, two ligand-one binding site Scatchard plot resulting from the presence of a constant concentration of competitive inhibitor is proposed; the same procedure may also be applied to the analysis of data derived from the "IC50" competitive inhibition experimental design. Furthermore, a useful generalization of the Cheng-Prusoff correction is presented.


Non-enzymatic post-translational protein modifications and proteostasis network deregulation in carcinogenesis.
Organisms are constantly challenged by stressors and thus the maintenance of biomolecules functionality is essential for the assurance of cellular homeostasis. Proteins carry out the vast majority of cellular functions by mostly participating in multimeric protein assemblies that operate as protein machines. Cells have evolved a complex proteome quality control network for the rescue, when possible, or the degradation of damaged polypeptides. Nevertheless, despite these proteostasis ensuring mechanisms, new protein synthesis, and the replication-mediated dilution of proteome damage in mitotic cells, the gradual accumulation of stressors during aging (or due to lifestyle) results in increasingly damaged proteome. Non-enzymatic post-translational protein modifications mostly arise by unbalanced redox homeostasis and/or high glucose levels and may cause disruption of proteostasis as they can alter protein function. This outcome may then increase genomic instability due to reduced fidelity in processes like DNA replication or repair. Herein, we present a synopsis of the major non-enzymatic post-translation protein modifications and of the proteostasis network deregulation in carcinogenesis. We propose that activation of the proteostasis ensuring mechanisms in premalignant cells has tumor-preventive effects, whereas considering that over-activation of these mechanisms represents a hallmark of advanced tumors, their inhibition provides a strategy for the development of anti-tumor therapies. This article is part of a Special Issue entitled: Posttranslational Protein modifications in biology and Medicine.


Long-term social and professional outcomes in adults after pediatric kidney failure.
Little is known about the long-term social and professional outcomes in adults after pediatric kidney replacement therapy (KRT). In this study, we described social and professional outcomes of adults after kidney failure during childhood and compared these outcomes with the general population. We sent a questionnaire to 143 individuals registered in the Swiss Pediatric Renal Registry (SPRR) with KRT starting before the age of 18 years. In the questionnaire, we assessed social (partner relationship, living situation, having children) and professional (education, employment) outcomes. Logistic regression models adjusted for age at study and sex were used to compare outcomes with a representative sample of the Swiss general population and to identify socio-demographic and clinical characteristics associated with adverse outcomes. Our study included 80 patients (response rate 56%) with a mean age of 39 years (range 19-63). Compared to the general population, study participants were more likely to not have a partner (OR = 3.7, 95%CI 2.3-5.9), live alone (OR = 2.5, 95%CI 1.5-4.1), not have children (OR = 6.8, 95%CI 3.3-14.0), and be unemployed (OR = 3.9, 95%CI 1.8-8.6). No differences were found for educational achievement (p = 0.876). Participants on dialysis at time of study were more often unemployed compared to transplanted participants (OR = 5.0, 95%CI 1.2-21.4) and participants with > 1 kidney transplantation more often had a lower education (OR = 3.2, 95%CI 1.0-10.2). Adults after pediatric kidney failure are at risk to experience adverse social and professional outcomes. Increased awareness among healthcare professionals and additional psycho-social support could contribute to mitigate those risks. A higher resolution version of the Graphical abstract is available as Supplementary information.


Ghrelin, leptin and insulin in healthy children: Relationship with anthropometry, gender, and age distribution.
This study aimed to establish the relationship between total ghrelin, acyl ghrelin, des-acyl ghrelin, leptin, and insulin with anthropometry, gender, and age distribution in healthy children. Data from 111 healthy children aged 4 months to 10 years were studied. All the participants underwent a pre-study screening clinical evaluation and were separated in 3 age groups. All had blood collected to assay. Anthropometric parameters were measured according to World Health Organization. In order to determine the correlation between dependent and independent variables, a multiple linear regression analysis was used. Overall median age of subjects was 60.0 months. After multiple regression analysis, correlation between total ghrelin, acyl ghrelin and des-acyl ghrelin remained significant with age. Correlation between leptin values and age, body mass index-for-age ratio, height-for-age ratio, and female gender remained significant. There was no significant correlation between insulin and ghrelin, and between insulin and leptin in all age groups. There was an inverse significant correlation between total ghrelin and des-acyl ghrelin with leptin in the whole group. Ghrelin showed an inverse correlation with age and leptin showed a direct correlation with anthropometric parameters and female gender in healthy children. Insulin did not show any correlation.


Effects of Pharmacologic Treatment for Neonatal Abstinence Syndrome on DNA Methylation and Neurobehavior: A Prospective Cohort Study.
To determine whether pharmacologic treatment for neonatal abstinence syndrome (NAS) is associated with changes in DNA methylation (DNAm) of the mu-opioid receptor gene (OPRM1) and improvements in neonatal neurobehavior. Buccal swabs were collected from 37 neonates before and after morphine treatment for NAS. Genomic DNA was extracted, and DNAm was examined at 4 cytosine-phosphate-guanine (CpG) sites within the OPRM1 gene. Assessment with the NICU Network Neurobehavioral Scales (NNNS) was also performed before and after NAS treatment. Changes in DNAm (DNAmpost-tx - DNAmpre-tx) and NNNS summary scores (NNNSpost-tx - NNNSpre-tx) were then calculated. Path analysis was used to examine associations among pharmacologic treatment (length of treatment [LOT] and total dose of morphine), changes in DNAm, and changes in NNNS summary scores. DNAm was significantly decreased from pretreatment to post-treatment at 1 of 4 CpG sites within the OPRM1 gene. Neonates also demonstrated decreased excitability, hypertonia, lethargy, signs of stress and abstinence, and increased quality of movement and regulation from pretreatment to post-treatment. Longer LOT and higher morphine dose were associated with greater decreases in DNAm; greater decreases in DNAm were associated with greater decreases in excitability and hypertonia on the NNNS. Pharmacologic treatment of NAS is associated with decreased DNAm of the OPRM1 gene and improved neonatal neurobehavior. Epigenetic changes may play a role in these changes in neonatal neurobehavior.


Complex modulation of human motor cortex excitability by the specific serotonin re-uptake inhibitor sertraline.
Monoamines are powerful modulators of cortical function. Serotonin has complex excitatory and inhibitory effects on animal cortex. Here, the effects of a single oral dose (100mg) of the selective serotonin re-uptake inhibitor sertraline on human motor cortex excitability were investigated in healthy subjects. Transcranial magnetic stimulation was used to test motor threshold, motor evoked potential intensity curve, cortical silent period, paired-pulse inhibition and facilitation and I-wave facilitation. Sertraline resulted in a steeper intensity curve and in depressed paired-pulse facilitation (PPF). All other measures and spinal and neuromuscular excitability remained unaffected. The steeper intensity curve points to an increased excitability of the cortico-spinal neurone, while the depressed PPF suggests an enhanced control of the cortico-spinal neurone by inhibitory interneurones. These features may improve the signal-to-noise ratio of output cells in human motor cortex.


A new on-axis micro-spectrophotometer for combining Raman, fluorescence and UV/Vis absorption spectroscopy with macromolecular crystallography at the Swiss Light Source.
The combination of X-ray diffraction experiments with optical methods such as Raman, UV/Vis absorption and fluorescence spectroscopy greatly enhances and complements the specificity of the obtained information. The upgraded version of the in situ on-axis micro-spectrophotometer, MS2, at the macromolecular crystallography beamline X10SA of the Swiss Light Source is presented. The instrument newly supports Raman and resonance Raman spectroscopy, in addition to the previously available UV/Vis absorption and fluorescence modes. With the recent upgrades of the spectral bandwidth, instrument stability, detection efficiency and control software, the application range of the instrument and its ease of operation were greatly improved. Its on-axis geometry with collinear X-ray and optical axes to ensure optimal control of the overlap of sample volumes probed by each technique is still unique amongst comparable facilities worldwide and the instrument has now been in general user operation for over two years.


Studies on aorta during development. II. Differences in ontogeny of the key enzymes involved in cholesteryl ester synthesis and hydrolysis in rabbit aorta.
It is well known that cholesteryl ester accumulation is dramatically increased in the atherosclerotic artery. The enzymes acyl-CoA: cholesterol acyltransferase (ACAT), acid cholesteryl esterase (ACE) and neutral cholesteryl esterase (NCE) may play key roles in the accumulation of cholesteryl esters in the arterial wall. However, very little is known regarding the developmental pattern of the key enzymes involved in cholesteryl ester synthesis and hydrolysis. The total activities of ACAT, ACE and NCE were measured by radioassay using liposomal substrates in rabbit aortic homogenates. Our results indicate that ACAT activity decreases as a quadratic function with age (P less than 0.05). ACAT activity (pmol/100 mg protein/min) decreased from a high value in the fetus at term (63.3 +/- 7.4) to gradually lower values with increasing age. On the other hand, ACE activity (pmol/mg protein/min) was low in the fetus at term, and changed as a quadratic function with age (P less than 0.05) increasing gradually to higher activities with age up to a maximum at 12 weeks then decreased at 21 weeks. NCE activity (pmol/mg protein/min) increased dramatically from a low value in the fetus at term (3.34 +/- 0.48) to a maximum value at 1.5 weeks (14.65 +/- 2.73) then decreased as a linear function with increasing age up to 21 weeks (P less than 0.05). Plasma total cholesterol (mg/dl) also increased sharply from the fetal value at term of 98.5 +/- 5.2 to a maximum value at 1.5 weeks of 666.4 +/- 33.4, then decreased as a quadratic function with increasing age up to 21 weeks (40.8 +/- 6.7) (P less than 0.05). The free cholesterol content (microgram/mg protein) of the aortic tissue was initially high in the fetus (24.8 +/- 5.9) then increased with age. Examination of the ratio of synthesis to hydrolysis of cholesteryl esters as an index of enzyme activity units demonstrated a very high index in the fetus of 6.1 that rapidly decreased with increasing age in the young adult rabbit down to a value of 0.4 by 21 weeks of age. Correlation coefficients between enzyme activities, plasma cholesterol levels and aortic cholesterol levels indicated (a) a positive correlation of NCE activity with plasma cholesterol, (b) a negative correlation of NCE and ACE with aortic-cholesteryl ester content, and (c) no significant correlation of ACAT activity with either plasma cholesterol or aortic cholesterol content, indicating other factors are involved.(ABSTRACT TRUNCATED AT 400 WORDS)


[Posterior knee instability].
Due to improved diagnostic methods, posterior knee instabilities are detected more often. Up to now, however, the natural history of isolated ruptures of the posterior cruciate ligament is not known exactly. After conservative treatment, the first degenerative changes occur after about 10-15 years, mainly in the femoropatellar joint and the medial compartment. In complex posterior instabilities with lesions of the secondary stabilizers, the posterolateral structures, the lateral collateral ligament, and the leg alignment play an important role. Conservative or operative treatment of posterior knee instabilities depends on the extent of the lesion, the violated structures, and the impairment of the patient. For replacement of the posterior cruciate ligament, grafts from the patella ligament, the quadriceps tendon, and the hamstrings are used most frequently. In simultaneous lesions of the posterolateral corner of the knee joint, additional ligament-stabilizing methods are necessary.


High prevalence of HIV-1 and hepatitis C virus coinfection among injection drug users in the southeastern region of Yunnan, China.
The southeastern region of Yunnan province is a key site for drug trafficking and HIV-1 infection spread from the west of Yunnan and Laos to southeastern China. To investigate the prevalence of HIV-1 infection and hepatitis C virus (HCV) coinfection among injection drug users (IDUs) in southeastern Yunnan, three cohorts of 285 addicts, including 242 IDUs and 43 oral drug users, living in the cities of Gejiu and Kaiyuan and the county of Yanshan were studied. HIV-1 and HCV infections were detected by enzyme-linked immunosorbent assay and/or polymerase chain reaction. Data on the age, sex, risk behavior, drug use history, employment, ethnic background, and marriage status were obtained by interview. The overall prevalence of HIV-1 infection was 71.9%. The rate of HCV coinfection among 138 HIV-1-infected IDUs was 99.3%. Most HIV-infected IDUs were 20 to 35 years old (86.7%) and were ethnic Han (75.9%), suggesting that the epidemic in Yunnan is no longer confined to non-Han ethnic minorities. HIV prevalence in female IDUs (81.2%) was significantly higher than in male IDUs (68.2%) ( p <.05). The prevalence of HIV infection reached 68.4% after 1 year of injection drug use. Needle/syringe sharing is the major high risk factor for the spread of HIV-1 and HCV infections. Large-scale educational campaigns are urgently needed to reduce the spread of HIV and HCV infection in these regions.


Transcriptional regulation of GSDML gene by antisense-oriented HERV-H LTR element.
During the course of hominoid evolution, a new transcript variant of the GSDML (gasdermin-like protein) gene was formed by the integration of the antisense-oriented HERV-H (human endogenous retrovirus) LTR (long terminal repeat) element. To investigate regions that are critical for transcriptional regulation of the GSDML gene, we generated seven deletion mutants from a full-length clone (clone 1/630) that includes the HERV-H LTR sequence and compared their expression levels relative to the full-length parental clone using a transient transfection assay. In the transient transfection assay, deletion of the 5' flanking region (cellular origin) of the HERV-H LTR sequence led to a 4.5-fold increase in expression compared to the full-length clone, while deletion of the U5 region showed a significant decrease in transcriptional activity. Deletion of the 3' flanking region of the LTR sequence (clone 42/451) showed similar transcriptional activity to a clone missing the 5' flanking region of cellular origin (clone 42/630). Taken together, these data indicate that the HERV-H LTR sequence (viral origin) positively regulates transcriptional activity of the GSDML gene and that the 5' flanking region sequence (cellular origin) exerts negative transcriptional regulation.


Efficacy of radiosynovectomy in rheumatoid arthritis.
In this retrospective study, we evaluated the effect of radiosynovectomy of patients with rheumatoid arthritis. Radiosynovectomy was performed in 577 joints of 137 rheumatoid patients. We applied 185 MBq yttrium-90 in knees (n = 58), 74-111 MBq rhenium-186 colloids in ankle (n = 50), wrists (n = 43) and shoulders (n = 35), and 15 to 37 MBq in finger (n = 298) and toe joints (n = 46). The effect of radiosynovectomy was scored in 4 subjective categories: excellent response (no symptoms); good response (significant reduction of symptoms); moderate response (slight decrease); and bad response (no change or worsening), of pain and/or swelling in treated joint 3 months after the procedure. Excellent or good response was observed in 57% of treated knees, 63% of shoulders, 60% of wrists, 64% of ankles, 54% of thumb bases, 55% of MCP's, 54% of PIP's, 53% of DIP's, and 54% of MTP's. Side effects associated to the RSO, i.e., swelling or transient increase of pain, were recorded in 7% of the patients that resolved within 1 month. No patient had any non-reversible skin alteration after treatment, only slight erythema was observed in 5 patients. Radiosynovectomy is effective and safe in the treatment of rheumatoid arthritis.


The chemical logic of plant natural product biosynthesis.
Understanding the logic of plant natural product biosynthesis is important for three reasons: it guides the search for new natural products and pathways, illuminates the function of existing pathways in the context of host biology, and builds an enabling 'parts list' for plant and microbial metabolic engineering. In this review, we highlight the chemical themes that underlie a broad range of plant pathways, dividing pathways into two parts: scaffold-generating steps that draw on a limited set of chemistries, and tailoring reactions that produce a wide range of end products from a small number of common scaffolds.


Synaptonemal complex karyotype of Eimeria tenella.
In most organisms, biological variability rests on the behaviour of the chromosomes in the meiotic context. Despite the importance of meiosis, very little is known about the meiotic behaviour of the Eimeria chromosomes. The aim of the present study is to describe the standard synaptonemal complex karyotype from Eimeria tenella oocyst spreads by electron microscopy. For that purpose, complete sets of pachytene synaptonemal complexes were obtained and the morphological pachytene karyotype was determined. The authors used a previously reported method that overcomes the difficulty of the extreme resistance of protozoan oocysts to disruption and permits the release of intact meiotic chromosomes. The chromosomes were selected under a light microscope and those selected were stained with phosphotungtic acid and studied by transmission electron microscopy. The authors confirmed 14 chromosomes, which were observed as synaptonemal complexes, and the karyotype was constructed by arranging synaptonemal complexes according to their relative lengths and kinetochore position. Components of the synaptonemal complex, lateral elements, central element, recombination nodules and kinetochore were observed. Measures of the kynetochore, width of the synaptonemal complex, diameter of the recombination nodule and length of the telomeres are given. Minimal and no significant differences were found between measures of chromosomes isolated from different Eimeria tenella strains. To the best of our knowledge, the present investigation for the first time identifies and describes the morphological characteristics of the synaptonemal complex of Eimeria tenella during the meiosis that occurs within the oocysts. In addition, the authors provide evidence of the presence of recombination nodules, suggesting that the recombination process may play an important role in the molecular evolution of this parasite.


Elongation factor 1 alpha from Saccharomyces cerevisiae. Rapid large-scale purification and molecular characterization.
Cytoplasmic elongation factor 1 alpha (EF-1 alpha) was purified to homogeneity from the yeast Saccharomyces cerevisiae using a large-scale procedure. The three steps of purification used were batch adsorption on phosphocellulose, phosphocellulose chromatography and, as the last step, GDP-Sepharose or Biorex column chromatography. The protein is very basic (pI = 9.2) and has an apparent molecular mass of 49 kDa, as determined by polyacrylamide gel electrophoresis using denaturing conditions. It is one of the most abundant proteins in yeast (about 5% of total soluble protein), as shown by two-dimensional gel electrophoresis and by immunological titration. A strong immunological and structural homology was found between yeast EF-1 alpha and elongation factors from other sources. Common immunological features were found between yeast and wheat germ EF-1 alpha. Tryptic hydrolysis of yeast EF-1 alpha in the presence of 25% glycerol generated a large trypsin-resistant polypeptide (Mr = 43,000) which had the same NH2-terminal sequence as the proteolyzed product from rabbit reticulocyte, Artemia salina EF-1 alpha and Escherichia coli EF-Tu. Completed DNA sequence determination of one structural gene for yeast EF-1 alpha confirmed a remarkable conservation of several protein sequence domains in yeast and animal EF-1 alpha (Cottrelle, P., Thiele, D., Price, V., Memet, S., Micouin, J.Y., Marck, C., Buhler, J.M. Sentenac, A., and Fromageot, P. (1985) J. Biol. Chem. 260, 3090-3096).


Turnover rate of metallothionein and cadmium in Mytilus edulis.
The results demonstrate the first attempt to determine metallothionein turnover in the whole soft tissues of mussels Mytilus edulis exposed to cadmium. Half-lives for metallothionein and cadmium are 25 and 300 days, respectively. As metallothionein degrades the released cadmium induces further synthesis of the protein, to which the metal becomes resequestered. The slow metallothionein turnover rates (compared with mammals) and the lack of significant cadmium excretion testify to the relatively stable nature of the cadmium-metallothionein complex in these invertebrates and supports the view of a detoxifying role for metallothionein in the mussels.


Dietary vitamin D intake and the bladder cancer risk: A pooled analysis of prospective cohort studies.
Diet may play an essential role in the aetiology of bladder cancer (BC). Vitamin D is involved in various biological functions which have the potential to prevent BC development. Besides, vitamin D also influences the uptake of calcium and phosphorus, thereby possibly indirectly influencing the risk of BC. The aim of the present study was to investigate the relation between vitamin D intake and BC risk. Individual dietary data were pooled from ten cohort studies. Food item intake was converted to daily intakes of vitamin D, calcium and phosphorus. Pooled multivariate hazard ratios (HRs), with corresponding 95% confidence intervals (CIs) were obtained using Cox-regression models. Analyses were adjusted for gender, age and smoking status (Model 1), and additionally for the food groups fruit, vegetables and meat (Model 2). Dose-response relationships (Model 1) were examined using a nonparametric test for trend. In total, 1994 cases and 518,002 non-cases were included in the analyses. The present study showed no significant associations between individual nutrient intake and BC risk. A significant decreased BC risk was observed for high vitamin D intake with moderate calcium and low phosphorus intake (Model 2: HRhigh vitD, mod Ca, low P: 0.77, 95% CI: 0.59-1.00). No significant dose-response analyses were observed. The present study showed a decreased BC risk for high dietary vitamin D intake in combination with low calcium intake and moderate phosphorus intake. The study highlights the importance of examining the effect of a nutrient in combination with complementary nutrients for risk assessment. Future research should focus on nutrients in a wider context and in nutritional patterns.


Omega-3 fatty acids decrease CRYAB, production of oncogenic prostaglandin E2 and suppress tumor growth in medulloblastoma.
Medulloblastoma (MB) is one of the most common malignant central nervous system tumors of childhood. Despite intensive treatments that often leads to severe neurological sequelae, the risk for resistant relapses remains significant. In this study we have evaluated the effects of the ω3-long chain polyunsaturated fatty acids (ω3-LCPUFA) docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) on MB cell lines and in a MB xenograft model. Effects of ω3-LCPUFA treatment of MB cells were assessed using the following: WST-1 assay, cell death probes, clonogenic assay, ELISA and western blot. MB cells were implanted into nude mice and the mice were randomized to DHA, or a combination of DHA and EPA treatment, or to control group. Treatment effects in tumor tissues were evaluated with: LC-MS/MS, RNA-sequencing and immunohistochemistry, and tumors, erythrocytes and brain tissues were analyzed with gas chromatography. ω3-LCPUFA decreased prostaglandin E2 (PGE2) secretion from MB cells, and impaired MB cell viability and colony forming ability and increased apoptosis in a dose-dependent manner. DHA reduced tumor growth in vivo, and both PGE2 and prostacyclin were significantly decreased in tumor tissue from treated mice compared to control animals. All ω3-LCPUFA and dihomo-γ-linolenic acid increased in tumors from treated mice. RNA-sequencing revealed 10 downregulated genes in common among ω3-LCPUFA treated tumors. CRYAB was the most significantly altered gene and the downregulation was confirmed by immunohistochemistry. Our findings suggest that addition of DHA and EPA to the standard MB treatment regimen might be a novel approach to target inflammation in the tumor microenvironment.


A comparison of depressed and anxious chronic pain patients.
This study compared 102 depressed pain patients and 41 anxious pain patients. Depressed pain patients were significantly more often older in age. Pain patients with anxiety disorder were more often younger and unemployed. There was no significant difference in the family psychiatric morbidity. Such a subdivision of chronic pain patients based on psychopathology could be useful for management purposes and for future research.


Effect of a hyperosmotic agent on epithelial disruptions during laser in situ keratomileusis.
To evaluate the incidence of epithelial disruptions during primary laser in situ keratomileusis (LASIK) with the use of a preoperative hyperosmotic treatment comprising sodium chloride 5% ophthalmic ointment (Muro-128) and to identify the incidence of epithelial disruptions in various demographic populations. TLC Laser Eye Center, McLean, Virginia, USA. Comparative case series. Using a matched-pair design, hyperosmotic treatment was randomized to 1 eye of patients having bilateral LASIK. The primary outcome measure, epithelial integrity, was assessed in each eye. Epithelial integrity was evaluated in groups defined by characteristics that included age, sex, ethnicity, skin type, presence or absence of rosacea, eye color, and hair color. The study evaluated 496 eyes of 248 patients. The preoperative hyperosmotic treatment was associated with significantly less corneal epithelial disruption, as indicated by an epithelial integrity score. Compared with control eyes, the rate of corneal epithelial disruptions in the population of treated eyes decreased by 40% (relative risk, 0.60; 95% confidence interval [CI], 0.38-0.95). Among the characteristics studied, age was the best predictor of corneal epithelial disruptions; every 1-year increase in age was associated with a 9.0% increase in the risk for corneal epithelial disruptions (odds ratio [OR], 1.09; 95% CI, 1.05-1.13). Eyes of patients older than 34 years had a 4.4 times greater odds of being associated with epithelial disturbances than eyes of patients 34 years or younger. The preoperative use of hyperosmotic agents can reduce the risk for intraoperative epithelial disruptions during LASIK. An increase in epithelial disruptions was associated with increasing age.


Detection of estrogen receptors ER-alpha and ER-beta in human ejaculated immature spermatozoa with excess residual cytoplasm.
A key role of estrogens in human sperm biology has been recently suggested by aromatase and estrogen receptor detection in human testicular germ cells and ejaculated spermatozoa. However, the involvement of these hormones in the sperm maturation process is still not defined. The aim of this work was to investigate the expression of estrogen receptors, ER-alpha and ER-beta, in human ejaculated immature spermatozoa with excess residual cytoplasm. Immunofluorescence labelling has been used to localize ER-alpha and ER-beta proteins in immature spermatozoa isolated by Percoll gradient, while Western blot analysis was carried out on sperm protein extracts. Both estrogen receptors were localized in excess residual cytoplasm of immature sperm, while sperm tails showed only ER-beta. Furthermore, in the same cells, immunoblots detected the presence of the full-length (approximately 67 kDa) ER-alpha and (approximately 59 kDa) ER-beta proteins, together with a approximately 50 kDa ER-beta species, lacking in mature sperm. The present investigation demonstrated ER-alpha and ER-beta presence in excess residual cytoplasm of human abnormal sperm cells, suggesting the hypothesis that both the 'classical' ERs could be able to mediate estrogen action in spermatogenetic cells. Furthermore, the presence of the short ER-beta form in abnormal germ cells and its disappearance in mature sperm, support estrogen modulation via different ER forms during sperm maturation.


Intradialytic Massage for Leg Cramps Among Hemodialysis Patients: a Pilot Randomized Controlled Trial.
Patients on hemodialysis often experience muscle cramps that result in discomfort, shortened treatment times, and inadequate dialysis dose. Cramps have been associated with adversely affecting sleep and health-related quality of life, depression and anxiety. There is limited evidence available about massage in dialysis; however, massage in cancer patients has demonstrated decreases in pain, inflammation, and feelings of anxiety. These correlations indicate massage may be an effective treatment modality for hemodialysis-related lower extremity cramping. To determine the effectiveness of intradialytic massage on the frequency of cramping among hemodialysis patients prone to lower extremity cramping. 26 maintenance hemodialysis patients with frequent lower extremity cramps. three outpatient hemodialysis centers in Northeast Ohio. randomized controlled trial. The intervention group received a 20-minute massage of the lower extremities during each treatment (three times per week) for two weeks. The control group received usual care by dialysis center staff. change in frequency of lower leg cramping. Patient reported cramping at home decreased by 1.3 episodes per week in the intervention group compared to 0.2 episodes per week in the control group (p=.005). Patient reported cramping during dialysis decreased by 0.8 episodes in the intervention group compared to 0.4 episodes in the control group (p=0.44). Intradialytic massage appears to be an effective way to address muscle cramping. Larger studies with longer duration should be conducted to further examine this approach.


The Role of Levodopa Challenge in Predicting the Outcome of Subthalamic Deep Brain Stimulation.
Deep brain stimulation of the subthalamic nucleus (STN-DBS) is an effective and evidence-based treatment for idiopathic Parkinson's disease (iPD). A minority of patients does not sufficiently benefit from STN-DBS. The predictive validity of the levodopa challenge for individual patients is analyzed. Data from patients assessed with a preoperative Levodopa-test and a follow-up examination (mean ± standard deviation: 9.15 months ±3.39) from Kiel (n = 253), Berlin (n = 78) and Toronto (n = 98) were studied. Insufficient DBS outcome was defined as an overall UPDRS-III reduction <33% compared to UPDRS-III in med-off at baseline or alternatively if the minimal clinically important improvement of 5 points was not reached. Single UPDRS-items and sub-scores were dichotomized. Following exploratory analysis, we trained supervised regression- and classification models for outcome prediction. Data analysis confirmed significant correlation between the absolute UPDRS-III reduction during Levodopa challenge and after stimulation. But individual improvement was inaccurately predicted with a large range of up to 30 UPDRS III points. Further analysis identified preoperative UPDRS-III/med-off-scores and preoperative Levodopa-improvement as most influential factors. The models for UPDRS-III and sub-scores improvement achieved comparably low accuracy. With large prediction intervals, the Levodopa challenge use for patient counseling is limited, though remains important for excluding non-responders to Levodopa. Despite these deficiencies, the current practice of patient selection is highly successful and builds not only on the Levodopa challenge. However, more specific motor tasks and further paraclinical tools for prediction need to be developed.


Effect of Jiawei Tongqiao Huoxue decoction in basilar artery dolichoectasia mice through yes-associated protein/transcriptional co-activator with PDZ-binding motif pathway.
The Jiawei Tongqiao Huoxue decoction (JTHD), composed of Acorus calamus var. angustatus Besser, Paeonia lactiflora Pall., Conioselinum anthriscoides 'Chuanxiong', Prunus persica (L.) Batsch, Ziziphus jujuba Mill., Carthamus tinctorius L., Pueraria montana var. lobata (Willd.) Maesen & S.M.Almeida ex Sanjappa & Predeep, Zingiber officinale Roscoe, Leiurus quinquestriatus, and Moschus berezovskii Flerov, was developed based on Tongqiao Huoxue decoction in Wang Qingren's "Yilin Gaicuo" in the Qing Dynasty. It has the effect of improving not only the blood flow velocity of vertebral and basilar arteries but also the blood flow parameters and wall shear stress. Especially in recent years, the potential efficacy of traditional Chinese medicine (TCM) for the treatment of basilar artery dolichoectasia (BAD) has attracted great attention as there are still no specific remedies for this disease. However, its molecular mechanism has not been elucidated. To identify the potential mechanisms of JTHD will help to intervene BAD and provide a reference for its clinical application. This study aims to establish a mouse model of BAD and explore the mechanism of JTHD regulating yes-associated protein/transcriptional co-activator with PDZ-binding motif (YAP/TAZ) pathway for attenuating BAD mice development. Sixty post-modeling C57/BL6 female mice were randomly divided into sham-operated, model, atorvastatin calcium tablet, low-dose JTHD, and high-dose JTHD groups. After 14 days of modeling, the pharmacological intervention was given for 2 months. Then, JTHD was analyzed by liquid chromatography-tandem mass spectrometry (LC-MS). ELISA was utilized to detect the changes in vascular endothelial growth factor (VEGF) and lipoprotein a (Lp-a) in serum. EVG staining was conducted to observe the pathological changes of blood vessels. TUNEL method was employed to detect the apoptosis rate of vascular smooth muscle cells (VSMCs). Micro-CT and ImagePro Plus software were used to observe and calculate the tortuosity index, lengthening index, percentage increase in vessel diameter, and tortuosity of the basilar artery vessels in mice. Western blot analysis was performed to detect the expression levels of YAP and TAZ proteins in the vascular tissues of mice. Many effective compounds such as choline, tryptophan, and leucine with anti-inflammation and vascular remodeling were identified in the Chinese medicine formula by LC-MS analysis. The serum levels of VEGF in the model mice decreased significantly while the levels of Lp-a increased obviously compared with those in the sham-operated group. The intima-media of the basilar artery wall showed severe disruption of the internal elastic layer, atrophy of the muscular layer, and hyaline changes of the connective tissue. Apoptosis of VSMCs added. Dilatation, elongation, and tortuosity of the basilar artery became notable, and tortuosity index, lengthening index, percentage increase in vessel diameter, and bending angle remarkably improved. The expression levels of YAP and TAZ protein in blood vessels elevated conspicuously (P < 0.05, P < 0.01). JTHD group markedly reduced the lengthening, bending angle, percentage increase in vessel diameter, and tortuosity index of basilar artery compared with the model group after 2 months of pharmacological intervention. The group also decreased the secretion of Lp-a and increased the content of VEGF. It inhibited the destruction of the internal elastic layer, muscular atrophy, and hyaline degeneration of connective tissue in basilar artery wall. The apoptosis of VSMCs was decreased, and the expression levels of YAP and TAZ proteins were abated (P < 0.05, P < 0.01). The mechanism of inhibition of basilar artery elongation, dilation, and tortuosity by JTHD, which has various anti-BAD effective compound components, may be related to the reduction in VSMCs apoptosis and downregulation of YAP/TAZ pathway expression.


[Preliminary studies of Conus geographus venom structural properties: evaluation of the toxin's molecular weight].
Venom from the posterior half of the venom duct of Conus Geographus was toxic to mice but without effect on a crab, and produced a flaccid spasmodic paralysis in mice. The minimum lethal dose for mice ranged from 5 micrograms to 50 micrograms (net weight) of venom per mouse. Toxin was submitted to Sephadex G 25 chromatography and to ultrafiltration. The venom was inactivated by pronase. The data suggested that the toxin was a peptide of molecular weight of about 1 500-2 000.


Prolonged allergen challenge in mice leads to persistent airway remodelling.
Inflammatory infiltrates, airway hyper-responsiveness, goblet cell hyperplasia and subepithelial thickening are characteristic of chronic asthma. Current animal models of allergen-induced airway inflammation generally concentrate on the acute inflammation following allergen exposure and fail to mimic all of these features. The aim of this study was to use a murine model of prolonged allergen-induced airway inflammation in order to characterize the cells and molecules involved in the ensuing airway remodelling. Moreover, we investigated whether remodelling persists in the absence of continued allergen challenge. Acute pulmonary eosinophilia and airways hyper-reactivity were induced after six serial allergen challenges in sensitized mice (acute phase). Mice were subsequently challenged three times a week with ovalbumin (OVA) (chronic phase) up to day 55. To investigate the persistence of pathology, one group of mice were left for another 4 weeks without further allergen challenge (day 80). The extended OVA challenge protocol caused significant airway remodelling, which was absent in the acute phase. Specifically, remodelling was characterized by deposition of collagen as well as airway smooth muscle and goblet cell hyperplasia. Importantly, these airway changes, together with tissue eosinophilia were sustained in the absence of further allergen challenge. Examination of cytokines revealed a dramatic up-regulation of IL-4 and tumour growth factor-beta1 during the chronic phase. Interestingly, while IL-4 levels were significantly increased during the chronic phase, levels of IL-13 fell. Levels of the Th1-associated cytokine IFN-gamma also increased during the chronic phase. In conclusion, we have demonstrated that prolonged allergen challenge results in persistent airway wall remodelling.


Measuring operational efficiency in a health care system: a case study from Thailand.
This paper investigates the economic relationship among medical resources and efficiency of the health care system in a developing Asian country. The rapid growth in the use of limited resources and the escalating national health expenditure, raise the critical economic question of whether the use of health care resources are efficient. We estimated a four-factor production system, based on 1982-1997 annual operational data comprising five cross-sectional regions per year. The translog production function and three derived demand for factor input equations were jointly estimated using systems regression method. Results show that different types of medical care workers (doctors, nurses, pharmacists) influenced efficiency differently. The marginal products (MPs) of nurses and capital are the highest and they varied across the regions. Third, the estimates of factor substitution possibilities indicate difficult factor adjustments; these estimates differ in magnitudes and significance across regions but they similarly classify all but one (different) input pair as economic substitutes. Fourth, the regional variations in returns to scale estimates in live births tend to converge to that of the Bangkok metropolis. Finally, technical change is physician and pharmacist labor using, but capital and nursing labor saving. Policy implications of these findings touch on Article 78 of the Thailand Constitution.


Ca(2+)-activated Cl(-) channels: a newly emerging anion transport family.
A new family of chloride transport proteins has recently emerged. These proteins have extensive homology to a protein previously isolated from bovine tracheal epithelium that acts as a Ca(2+)-sensitive Cl(-) channel (CaCC) when heterologously expressed or when reconstituted into planar lipid bilayers. Several new members of this family have been identified in human, murine, and bovine epithelia, in addition to some other tissues, and are associated with Ca(2+)-sensitive conductive chloride transport when heterologously expressed in Xenopus oocytes or HEK 293 cells. The expressed current is also sensitive to inhibitors such as DIDS and niflumic acid. In addition, at least one family member acts as an endothelial cell adhesion molecule. This emerging family may underlie the Ca(2+)-mediated Cl(-) conductance responsible for rescue of the cystic fibrosis (CF) knockout mouse from significant airway disease.


Ecotoxicogenomics: the challenge of integrating genomics into aquatic and terrestrial ecotoxicology.
Rapid progress in the field of genomics (the study of how an individual's entire genetic make-up, the genome, translates into biological functions) is beginning to provide tools that may assist our understanding of how chemicals can impact on human and ecosystem health. In many ways, if scientific and regulatory efforts in the 20th century have sought to establish which chemicals cause damage to ecosystems, then the challenge in ecotoxicology for the 21st century is to understand the mechanisms of toxicity to different wildlife species. In the human context, 'toxicogenomics' is the study of expression of genes important in adaptive responses to toxic exposures and a reflection of the toxic processes per se. Given the parallel implications for ecological (environmental) risk assessment, we propose the term 'ecotoxicogenomics' to describe the integration of genomics (transcriptomics, proteomics and metabolomics) into ecotoxicology. Ecotoxicogenomics is defined as the study of gene and protein expression in non-target organisms that is important in responses to environmental toxicant exposures. The potential of ecotoxicogenomic tools in ecological risk assessment seems great. Many of the standardized methods used to assess potential impact of chemicals on aquatic organisms rely on measuring whole-organism responses (e.g. mortality, growth, reproduction) of generally sensitive indicator species at maintained concentrations, and deriving 'endpoints' based on these phenomena (e.g. median lethal concentrations, no observed effect concentrations, etc.). Whilst such phenomenological approaches are useful for identifying chemicals of potential concern they provide little understanding of the mechanism of chemical toxicity. Without this understanding, it will be difficult to address some of the key challenges that currently face aquatic ecotoxicology, e.g. predicting toxicant responses across the very broad diversity of the phylogenetic groups present in aquatic ecosystems; estimating how changes at one ecological level or organisation will affect other levels (e.g. predicting population-level effects); predicting the influence of time-varying exposure on toxicant responses. Ecotoxicogenomic tools may provide us with a better mechanistic understanding of aquatic ecotoxicology. For ecotoxicogenomics to fulfil its potential, collaborative efforts are necessary through the parallel use of model microorganisms (e.g. Saccharomyces cerevisiae) together with aquatic (e.g. Danio rerio, Daphnia magna, Lemna minor and Xenopus tropicalis) and terrestrial (e.g. Arabidopsis thailiana, Caenorhabdites elegans and Eisenia foetida) plants, animals and microorganisms.


Response to an experimental HBV vaccine permits withdrawal of HBIg prophylaxis in fulminant and selected chronic HBV-infected liver graft recipients.
Strategies using lamivudine and hepatitis B immunoglobulins (HBIg) for prevention of hepatitis B virus (HBV) reinfection after liver transplantation (LT) are expensive since life-long treatment is needed. We evaluated the possibility to obtain protective hepatitis B surface antigen (HBsAg) antibody (anti-HBs) titers after LT and to discontinue HBIg prophylaxis after a reinforced course of vaccination against HBV using an experimental adjuvant HBsAg / AS04 vaccine (GlaxoSmithKline Biologicals [GSK], Rixensart, Belgium) in patients transplanted for hepatitis B. Fifteen LT patients on stable low-level immunosuppression were vaccinated with a double dose of the vaccine at 0, 1, 2, 6, and 12 months: 5 patients were transplanted for nonviral diseases and 10 patients were transplanted for HBV on HBIg monotherapy. HBIg were continued during baseline vaccination (0, 1, and 2 months) and when anti-HBs titers determined every 6 weeks dropped below 150 IU/L. Overall follow-up was 18 months. Sustained long-term response to vaccination was defined as anti-HBs titers >500 IU/L without further need for HBIg administration during a follow-up period of at least 12 months. Overall sustained response to vaccination was 53% (8 / 15 patients); 80% (4 / 5 patients) in the nonviral disease group and 40% (4 / 10 patients) in the HBV group (2 /2 fulminant and 2/8 chronically infected patients) developed a sustained long-term response and were completely free of HBIg at the end of the 18-month follow-up. No HBV recurrence, rejection episodes, or side effects occurred during the follow-up. In conclusion, protective anti-HBs titers were obtained in a substantial number of LT patients following a reinforced course of HBV vaccination with vaccines containing new immunostimulating adjuvants. Vaccination seems well tolerated and safe and allows long-term discontinuation of HBIg.


Accuracy of SARC-F and SARC-CalF for sarcopenia screening in older women from southern Brazil.
The aim of this study was to verify the accuracy of the SARC-F and the SARC-CalF as screening tools for sarcopenia in community-dwelling older women ≥60 y of age. This was a cross-sectional study evaluating a convenience sample of women ≥60 y of age, living in Southern Brazil. Sarcopenia was defined according to the criteria proposed in the latest European Working Group on Sarcopenia in Older People consensus (EWGSOP2). Appendicular muscle mass was assessed by dual-energy x-ray absorptiometry. Muscle strength was measured by handheld dynamometry, and physical performance through the 4-m gait speed test. The SARC-F questionnaire and SARC-CalF score for sarcopenia screening were also applied. We evaluated 288 participants, with a mean age of 67.6 ± 5.8 y. The frequency of probable and confirmed sarcopenia in the sample was 7.3% and 2.1%, respectively. The frequency of risk for sarcopenia assessed by the SARC-F was 4.5% and SARC-CalF 22.2%. Despite the excellent specificity (95.4%) demonstrated by the SARC-F, its sensitivity in identifying confirmed cases was null, whereas the SARC-CalF showed high sensitivity (83.3%) and good specificity (79%). The present study findings suggested that SARC-CalF may be able to outperform SARC-F as a sarcopenia screening tool in women ≥60 y of age even under the new EWGSOP2 criteria, the main determinant of which is strength as observed in studies based on the previous definition.


Polymorphisms in oxidative stress, metabolic detoxification, and immune function genes, maternal exposure to ambient air pollution, and risk of preterm birth in Taiyuan, China.
Exposure to air pollutants may be associated with preterm birth (PB) through oxidative stress, metabolic detoxification, and immune system processes. However, no study has investigated the interactive effects of maternal air pollution and genetic polymorphisms in these pathways on risk of PB. The study included 126 PB and 310 term births. A total of 177 single nucleotide polymorphisms (SNPs) in oxidative stress, immune function, and metabolic detoxification-related genes were examined and analyzed. The China air quality index (AQI) was used as an overall estimation of ambient air pollutants. Among 177 SNPs, four SNPs (GPX4-rs376102, GLRX-rs889224, VEGFA-rs3025039, and IL1A-rs3783550) were found to have significant interactions with AQI on the risk of PB (Pinteraction were 0.001, 0.003, 0.03, and 0.04, respectively). After being stratified by the maternal genotypes in these four SNPs, 1.38 to 1.76 times of the risk of PB were observed as per interquartile range increase in maternal AQI among women who carried the GPX4-rs376102 AC/CC genotypes, the GLRX-rs889224 TT genotype, the VEGFA-rs3025039 CC genotype, or the IL1A-rs3783550 GT/TT genotypes. After adjustment for multiple comparisons, only GPX4-rs376102 and AQI interaction remained statistically significant (false discovery rate (FDR)=0.17). After additional stratification by preeclampsia (PE) status, a strongest association was observed in women who carried the GPX4-rs376102 AC/CC genotypes (OR, 2.26; 95% CI, 1.41-3.65, Pinteraction=0.0002, FDR=0.035) in the PE group. Our study provided the first evidence that association between maternal air pollution and PB risk may be modified by the genetic polymorphisms in oxidative stress and immune function genes. Future large studies are necessary to replicate and confirm the observed associations.


Family perspectives on end-of-life care experiences in nursing homes.
The purpose of this study is to expand knowledge regarding end-of-life care received in nursing homes through the use of narrative interviews with family members close to the decedents. We conducted follow-up qualitative interviews with 54 respondents who had participated in an earlier national survey of 1,578 informants. Interviews were taped and transcribed and then coded by a five-member, multidisciplinary team to identify overarching themes. Respondents report that the needs of dying patients are often insufficiently addressed by health care professionals. Their low expectations of nursing homes and their experiences cause many to become vigilant advocates. Respondents report that physicians are often "missing in action," and they desire more and better trained staff. They indicate that regulations reinforce task-focused rather than person-centered care and add to patient and family burden. Although hospice services are reported to enhance end-of-life care, respondents also report late referrals and occasional misunderstandings about the role and scope of hospice. Sustained efforts on many fronts are needed to improve end-of-life care in nursing homes. Policy recommendations are suggested.


Simple and efficient cell disruption of extremely small quantities of mycelium of phytopathogenic mycotoxin-producing moulds for quantitative extraction of genomic DNA.
DNA was efficiently and quantitatively isolated from extremely small quantities of mycelia (0.1-10 mg) of different phytopathogenic moulds by grinding freeze-dried mycelia with glass beads and then using a commercial DNA extraction kit. The efficiency of disruption of the mycelia and the quantitative DNA extraction was proved by microscopy and the quantification of isolated DNA by real time PCR.


Subperiosteal onlay augmentation of the mandible: a clinical and radiographic survey.
A five-year follow-up study of 26 patients who underwent subperiosteal onlay augmentation of the mandible using corticocancellous bone from the iliac crest is presented. An overall postoperative reduction in mandibular height of 44% was found. The different rates of resorption between the various suggested augmentation methods are discussed, and suggestions are given to prevent the rapid resorption that occurs during the immediate postoperative period.


[Familial Hodgkin's disease: a case report and review of the literature].
In a family of five sisters four developed malignant lymphomas. A case report is given together with a literature survey of familial Hodgkin's disease. The observations fit the hypothesis that for the development of Hodgkin's disease two requirements must be met: a genetically determined susceptibility as well as some causative agent.


Protein determination in materials containing melanin.
Strategy of protein determination in melanin containing samples was analyzed, and the prerequisites under which the Lowry procedure could be used were established. (a) Complete solubilization of proteins from melanosomes was not reached until the treatment with 3% sodium dodecylsulfate at 100 degrees C was prolonged to 5 h. (b) It is necessary to correct the data obtained with Folin reaction to eliminate melanin interference. (c) The correction is based upon the determination of the extent of solubilized melanin and subtraction of the corresponding Folin-Ciocalteu reaction values. (d) The extent of correction due to the melanin interference was found to be about 40% of the absorbance value. It was concluded that correction of the data obtained with Lowry assay is possible, but the procedure is complicated and time-consuming. However, a more simple modification has not been developed.


Reference values of the carotid elastic modulus using shear wave elastography and arterial stiffness change in coronary slow flow.
Shear wave elastography (SWE) accurately and sensitively evaluates arterial wall stiffness by quantifying the elastic modulus (EM); however, the absence of reference values has precluded its widespread clinical application. This prospective cohort study aimed to establish reference values for the carotid EM using SWE; investigate the main determinants of the EM; and evaluate EM changes in coronary slow flow (CSF), which is characterized by delayed coronary opacification without evident obstructive lesion in epicardial coronary artery on angiography. This study enrolled 169 healthy volunteers and 30 patients with CSF. The carotid maximum EM (EMmax), mean EM, and minimum EM were measured using SWE. CSF was diagnosed by thrombolysis in the myocardial infarction frame count during coronary angiography. No differences were found in the EM between the left and right carotid arteries and between men and women. Multiple linear regression analysis revealed that age was independently correlated with the EMmax, which progressively increased with age. Moreover, smoking had an independent influence on the EM after adjusting for age; smokers had higher EM than non-smokers. Age-specific reference values for the carotid EM were established. The EM was higher in patients with CSF than in controls after adjusting for age and smoking status. This study first established the reference values for the carotid EM using SWE. Age and smoking status were the main determinants of the EM. Patients with CSF had high EM. SWE can effectively and noninvasively evaluate arterial stiffness in patients with CSF.


A chestnut seed cystatin differentially effective against cysteine proteinases from closely related pests.
Cystatin CsC, a cysteine proteinase inhibitor from chestnut (Castanea sativa) seeds, has been purified and characterized. Its full-length cDNA clone was isolated from an immature chestnut cotyledon library. The inhibitor was expressed in Escherichia coli and purified from bacterial extracts. Identity of both seed and recombinant cystatin was confirmed by matrix-assisted laser desorption/ionization mass spectrometry analysis, two-dimensional electrophoresis and N-terminal sequencing. CsC has a molecular mass of 11,275 Da and pI of 6.9. Its amino acid sequence includes all three motifs that are thought to be essential for inhibitory activity, and shows significant identity to other phytocystatins, especially that of cowpea (70%). Recombinant CsC inhibited papain (Ki 29 nM), ficin (Ki 65 nM), chymopapain (Ki 366 nM), and cathepsin B (Ki 473 nM). By contrast with most cystatins, it was also effective towards trypsin (Ki 3489 nM). CsC is active against digestive proteinases from the insect Tribolium castaneum and the mite Dermatophagoides farinae, two important agricultural pests. Its effects on the cysteine proteinase activity of two closely related mite species revealed the high specificity of the chestnut cystatin.


Evaluation of the intra- and inter-specific genetic variability of Plasmodium lactate dehydrogenase.
Malaria diagnosis is vital to efficient control programmes and the recent advent of malaria rapid diagnostic tests (RDTs) provides a reliable and simple diagnostic method. However a characterization of the efficiency of these tests and the proteins they detect is needed to maximize RDT sensitivity. Plasmodial lactate dehydrogenase (pLDH) gene of wild isolates of the four human species of Plasmodium from a variety of malaria endemic settings were sequenced and analysed. No variation in nucleotide was found within Plasmodium falciparum, synonymous mutations were found for Plasmodium malariae and Plasmodium. vivax; and three different types of amino acid sequence were found for Plasmodium ovale. Conserved and variable regions were identified within each species. The results indicate that antigen variability is unlikely to explain variability in performance of RDTs detecting pLDH from cases of P. falciparum, P. vivax or P. malariae malaria, but may contribute to poor detection of P. ovale.


Assessment of physical functioning in patients with chronic obstructive pulmonary disease (COPD) requiring long-acting dual bronchodilation in routine clinical practice.
We aim to determine the effect of a fixed-dose combination (FDC) of tiotropium/olodaterol on Physical activity (PA) in patients with chronic obstructive pulmonary disease (COPD) in a real world setting. COPD patients were prospectively enrolled to evaluate the effect of a FDC of tiotropium/olodaterol inhaler therapy via the Respimat® Soft Mist™ inhaler (SMI) on the physical functioning scale (PF-10), and the general condition of the patient as assessed by the physician (Physician's Global Evaluation, PGE), and the patient's satisfaction after 6 weeks of treatment. The primary end-point was the percentage of patients with therapeutic success at 6th week follow-up, defined as a ≥10-points increase in the standardised PF-10 score from baseline. A total of 257 patients from 57 sites were enrolled, and 234 completed the follow up. After 6 weeks of treatment, 155 out of 234 patients (66.2%) showed therapeutic success in the physical functioning score, coupled with significant improvement in PGE score: 78 (33.3%) patients with good/excellent PGE score at baseline, increasing to 172 (73.5%) at 6th week (p<0.0001). The patient's satisfaction was excellent: 77.2% reporting to be satisfied/very satisfied with the treatment, 79.9% with inhaling and 79.0% with the handling of SMI device. 1.6% of patients reported an investigator-defined drug-related adverse event. Treatment of COPD patients with a FDC of tiotropium/olodaterol SMI for 6 weeks resulted in significant improvements in the patients' condition as assessed by patients and physicians, with no new safety findings.


GlideScope videolaryngoscope-assisted retrieval of an intratracheal foreign body.
Postintubation tracheal stenosis presents major challenges to the anesthesiologist, especially in situations where the airway is shared with the surgeon. The airway management of a patient with severe postintubation subglottic stenosis, who developed complete airway obstruction during attempted tracheal dilatation, is presented.


The plain chest radiograph after acute smoke inhalation.
Serial chest radiographs of 45 patients from a major fire disaster were assessed for the effects of smoke inhalation injury. Thirty-three had abnormal chest radiographs on admission. Bronchial wall thickening was present in a total of 29 patients, subglottic oedema in 13, pulmonary oedema in seven, and patchy consolidation in three. Seven patients developed pulmonary oedema following initial exposure. This was the cause of death in two patients. Serial chest radiographs were useful in monitoring this very severe complication. Bronchial wall thickening and subglottic oedema were common early findings and thus may herald more serious subsequent respiratory embarrassment. Subglottic oedema is a previously undescribed finding. It not only indicates damage to the upper airways but may also indicate damage to the lung parenchyma. It may also make endotracheal intubation difficult. Therefore, the initial chest radiograph is an important predictor of significant smoke inhalation injuries enabling selection of patients likely to need ventilatory support.