The process of implementing a rural VA wound care program for diabetic foot ulcer patients.
Delivering and documenting evidence-based treatment to all Department of Veterans Affairs (VA) foot ulcer patients has wide appeal. However, primary and secondary care medical centers where 52% of these patients receive care are at a disadvantage given the frequent absence of trained specialists to manage diabetic foot ulcers. A retrospective review of diabetic foot ulcer patient records and a provider survey were conducted to document the foot ulcer problem and to assess practitioner needs. Results showed of the 125 persons with foot ulcers identified through administrative data, only, 21% of diabetic foot patients were correctly coded. Chronic Care and Microsystem models were used to prepare a tailored intervention in a VA primary care medical center. The site Principal Investigators, a multidisciplinary site wound care team, and study investigators jointly implemented a diabetic foot ulcer program. Intervention components include wound care team education and training, standardized good wound care practices based on strong scientific evidence, and a wound care template embedded in the electronic medical record to facilitate data collection, clinical decision making, patient ordering, and coding. A strategy for delivering offloading pressure devices, regular case management support, and 24/7 emergency assistance also was developed. It took 9 months to implement the model. Patients were enrolled and followed for 1 year. Process and outcome evaluations are on-going.


S-adenosylmethionine inhibits the growth of cancer cells by reversing the hypomethylation status of c-myc and H-ras in human gastric cancer and colon cancer.
A global DNA hypomethylation might activate oncogene transcription, thus promoting carcinogenesis and tumor development. S-Adenosylmethionine (SAM) serves as a major methyl donor in biological transmethylation events. The object of this study is to explore the influence of SAM on the status of methylation at the promoter of the oncogenes c-myc, H-ras and tumor-suppressor gene p16 (INK4a), as well as its inhibitory effect on cancer cells. The results indicated that SAM treatment inhibited cell growth in gastric cancer cells and colon cancer cells, and the inhibition efficiency was significantly higher than that in the normal cells. Under standard growth conditions, C-myc and H-ras promoters were hypomethylated in gastric cancer cells and colon cancer cells. SAM treatment resulted in a heavy methylation of these promoters, which consequently downregulated mRNA and protein levels. In contrast, there was no significant difference in mRNA and protein levels of p16 (INK4a) with and without SAM treatment. SAM can effectively inhibit the tumor cells growth by reversing the DNA hypomethylation on promoters of oncogenes, thus down-regulating their expression. With no influence on the expression of the tumor suppressor genes, such as P16, SAM could be used as a potential drug for cancer therapy.


Car Driving Status and Living Arrangement Associated with Sarcopenia among Rural Japanese Older Adults: A Cross-Sectional Study.
Ensuring mobility after driving cessation is an important public health issue to prevent functional limitations, but this issue is still not fully understood in rural settings. The aim of this study was to test the hypothesis that being a non-driver and living alone is associated with a greater risk of sarcopenia among the community-dwelling elderly in rural Japanese areas. This study was conducted in 2018 and data from 738 participants were used. Sarcopenia was assessed by measuring walking speed, handgrip strength, and skeletal muscle mass. Car driving status and living arrangement were collected using self-reported questionnaires and face-to-face interviews. Four groups were set to determine combined conditions of car driving status and living arrangement. Logistic regression analysis was performed to estimate the odds ratio (OR) and a 95% confidence interval of sarcopenia after adjustment for confounding factors. Compared with the reference group (driver and living with others), the OR of sarcopenia was significantly higher in the non-driver and living alone group (OR = 2.21; 95% confidence interval, 1.02-4.80). Our findings suggest that the consideration of both driving status and living arrangement are important in the formulation of public health strategies to prevent sarcopenia in rural Japanese areas.


Improved neurological function in a patient with complete spinal cord injury through bone marrow mesenchymal stem cells implantation: a case report.
Aim: Spinal cord injury (SCI) can cause severe disability. Several clinical trials of stem-cell based therapies are ongoing. We describe our experience of bone marrow mesenchymal stem cell (BMSC) therapy in a patient with complete SCI in the chronic stage. Case report: A 25-year-old man with complete SCI at T6 level presented with paraplegia for 5 years. We transplanted autologous BMSCs intramedullary. After 12 months follow-up, his Barthel index score was noticeably improved from severe to moderate dependency, and the sensation level improved from T7 to S5, but no improvement of motor function. Conclusion: Autologous BMSCs are potentially safe for patients with complete SCI in the chronic stage and may improve neurological function and quality of life.


Monitoring of airborne contamination during the handling of technetium-99m and radioiodine.
Measurements have been made using an air sampler to measure airborne radioactivity produced during the routine handling of large activities of technetium-99m, iodine-125 and iodine-131. The results indicate that 99Tcm can be safely handled in environments without direct exhaust of the ventilated air but that 125I and 131I should always be handled in a ventilated environment such as a fume cupboard or a down-draught work-station of the total-exhaust type. Monitoring of thyroid uptake proves to be the most reliable means of monitoring airborne contamination by these radionuclides, but burdens and radiation doses for typical procedures are well within the maximum permissible limits of the Code of Practice for the Protection of Persons from Ionizing Radiation arising from Medical and Dental Use.


Aminobacter MSH1-Mineralisation of BAM in Sand-Filters Depends on Biological Diversity.
BAM (2,6-dichlorobenzamide) is a metabolite of the pesticide dichlobenil. Naturally occurring bacteria that can utilize BAM are rare. Often the compound cannot be degraded before it reaches the groundwater and therefore it poses a serious threat to drinking water supplies. The bacterial strain Aminobacter MSH1 is a BAM degrader and therefore a potential candidate to be amended to sand filters in waterworks to remediate BAM polluted drinking water. A common problem in bioremediation is that bacteria artificially introduced into new diverse environments often thrive poorly, which is even more unfortunate because biologically diverse environments may ensure a more complete decomposition. To test the bioaugmentative potential of MSH1, we used a serial dilution approach to construct microcosms with different biological diversity. Subsequently, we amended Aminobacter MSH1 to the microcosms in two final concentrations; i.e. 10(5) cells mL(-1) and 10(7) cells mL(-1). We anticipated that BAM degradation would be most efficient at "intermediate diversities" as low diversity would counteract decomposition because of incomplete decomposition of metabolites and high diversity would be detrimental because of eradication of Aminobacter MSH1. This hypothesis was only confirmed when Aminobacter MSH1 was amended in concentrations of 10(5) cells mL(-1). Our findings suggest that Aminobacter MSH1 is a very promising bioremediator at several diversity levels.


Structures of human cytochrome P-450 2E1. Insights into the binding of inhibitors and both small molecular weight and fatty acid substrates.
Human microsomal cytochrome P-450 2E1 (CYP2E1) monooxygenates > 70 low molecular weight xenobiotic compounds, as well as much larger endogenous fatty acid signaling molecules such as arachidonic acid. In the process, CYP2E1 can generate toxic or carcinogenic compounds, as occurs with acetaminophen overdose, nitrosamines in cigarette smoke, and reactive oxygen species from uncoupled catalysis. Thus, the diverse roles that CYP2E1 has in normal physiology, toxicity, and drug metabolism are related to its ability to metabolize diverse classes of ligands, but the structural basis for this was previously unknown. Structures of human CYP2E1 have been solved to 2.2 angstroms for an indazole complex and 2.6 angstroms for a 4-methylpyrazole complex. Both inhibitors bind to the heme iron and hydrogen bond to Thr303 within the active site. Complementing its small molecular weight substrates, the hydrophobic CYP2E1 active site is the smallest yet observed for a human cytochrome P-450. The CYP2E1 active site also has two adjacent voids: one enclosed above the I helix and the other forming a channel to the protein surface. Minor repositioning of the Phe478 aromatic ring that separates the active site and access channel would allow the carboxylate of fatty acid substrates to interact with conserved 216QXXNN220 residues in the access channel while positioning the hydrocarbon terminus in the active site, consistent with experimentally observed omega-1 hydroxylation of saturated fatty acids. Thus, these structures provide insights into the ability of CYP2E1 to effectively bind and metabolize both small molecule substrates and fatty acids.


Sympathetic inhibition, leptin, and uncoupling protein subtype expression in normal fasting rats.
To further investigate neural effects on leptin and uncoupling proteins (UCPs), we studied in vivo perturbations intended to block adrenergic input to peripheral tissues. We examined plasma leptin, leptin mRNA, and adipose and muscle UCP subtype mRNA in rats treated with alpha-methyl-p-tyrosine methyl ester (AMPT-ME), which inhibits catecholamine synthesis and 6-hydroxydopamine (6HDA), which is toxic to catecholinergic nerve terminals but, unlike AMPT-ME, does not enter the central nervous system. Intraperitoneal AMPT-ME, 250 mg/kg, was administered at 1800 and 0700 the following day, and rats were killed at 1200-1400. All rats were fasted with free access to water during this time. Intraperitoneal AMPT-ME increased plasma leptin by 15-fold, increased interscapular brown adipose tissue (IBAT) and epididymal fat leptin mRNA by 2- to 2.5-fold, and also increased plasma insulin and glucose concentrations. Intraperitoneal AMPT-ME decreased IBAT UCP-3 mRNA to 40% of control, while it increased epididymal adipose UCP-3 mRNA approximately twofold. Intravenous AMPT-ME, 250 mg/kg, administered to conscious rats for 5 h decreased lumbar sympathetic nerve activity, increased plasma leptin (5.89 +/- 1.43 compared with 2.75 +/- 0.31 ng/ml in vehicle-treated rats, n = 7, P < 0.05), and decreased cardiac rate with no sustained change in blood pressure. Intraperitoneal 6HDA, 100 mg/kg, as a single dose at 1800, increased plasma leptin approximately twofold after 18-20 h, increased IBAT (but not epididymal fat) leptin mRNA by two- to threefold, and decreased IBAT UCP-3 mRNA to 30-40% of control. Neither AMPT-ME nor 6HDA significantly altered mRNA encoding gastrocnemius muscle UCP-3, IBAT UCP-1, or IBAT and epididymal UCP-2. In summary, AMPT-ME and 6HDA increased plasma leptin and upregulated leptin mRNA expression. AMPT-ME also resulted in complex tissue and subtype-specific modulation of adipose UCP mRNA. These data are consistent with interaction between leptin and sympathetic nerve activity (SNA) in regulation of fat cell energy utilization. However, the in vivo modulation of leptin and UCPs appears complex and, beyond a causal effect of SNA per se, may depend on concurrent changes in plasma insulin, glucose, and circulatory hemodynamics.


The Expanding Spectrum of Primary Aldosteronism: Implications for Diagnosis, Pathogenesis, and Treatment.
Primary aldosteronism is characterized by aldosterone secretion that is independent of renin and angiotensin II and sodium status. The deleterious effects of primary aldosteronism are mediated by excessive activation of the mineralocorticoid receptor that results in the well-known consequences of volume expansion, hypertension, hypokalemia, and metabolic alkalosis, but it also increases the risk for cardiovascular and kidney disease, as well as death. For decades, the approaches to defining, diagnosing, and treating primary aldosteronism have been relatively constant and generally focused on detecting and treating the more severe presentations of the disease. However, emerging evidence suggests that the prevalence of primary aldosteronism is much greater than previously recognized, and that milder and nonclassical forms of renin-independent aldosterone secretion that impart heightened cardiovascular risk may be common. Public health efforts to prevent aldosterone-mediated end-organ disease will require improved capabilities to diagnose all forms of primary aldosteronism while optimizing the treatment approaches such that the excess risk for cardiovascular and kidney disease is adequately mitigated. In this review, we present a physiologic approach to considering the diagnosis, pathogenesis, and treatment of primary aldosteronism. We review evidence suggesting that primary aldosteronism manifests across a wide spectrum of severity, ranging from mild to overt, that correlates with cardiovascular risk. Furthermore, we review emerging evidence from genetic studies that begin to provide a theoretical explanation for the pathogenesis of primary aldosteronism and a link to its phenotypic severity spectrum and prevalence. Finally, we review human studies that provide insights into the optimal approach toward the treatment of primary aldosteronism.


Site selection for heart failure clinical trials in the USA.
There are more than 1 million primary hospitalizations for heart failure (HF) annually in the USA alone, and post-discharge outcomes remain persistently poor despite available therapies and quality improvement initiatives. Recent international randomized clinical trials in hospitalized HF have repeatedly failed to improve this post-discharge event rate. A potential reason for this persistent lack of clinical trial success that has not previously received significant attention relates to site selection and the generally low level of patient enrollment from the USA. Only ~5 % of US hospitals participate in clinical trials, and in four recent randomized trials of hospitalized HF, only one-third of patients were enrolled in North America. This poor participation among US centers has necessitated disproportionate enrollment from non-US sites. Regional variations in HF patient characteristics and clinical outcomes are well documented, and a lack of US patient representation in clinical trials limits the generalizability of results and presents obstacles for US regulatory agency approval. There are multiple impediments to successful US enrollment including a lack of incentive for investigators and institutions, the relative value unit-based compensation system, poor institutional framework for identification of appropriate patients, and increasing liability to conduct trials. In this manuscript, we specifically identify barriers to successful hospitalized HF clinical trial participation in the USA and suggest possible solutions.


Study of early atherosclerotic markers in women with polycystic ovary syndrome.
Women with polycystic ovary syndrome (PCOS) may represent a large underappreciated segment of female population who is at increased cardiovascular risk because of the presence of cluster of metabolic abnormalities. The aim of our study was to assess atherosclerotic risk factors in women with PCOS. In a cross-sectional study, 50 women with PCOS and 50 age and weight-matched healthy controls were enrolled. Endothelial dysfunction by flow-mediated dilatation (FMD) of brachial artery, highly sensitive C-reactive protein (hs CRP), and carotid intima media thickness (CIMT) were measured in both cases and control groups. The mean age of women with PCOS was 26.82 ± 3.26 years and Body-mass index (BMI) of 26.2 ± 4.8 kg/ m(2). Thirty-six (72%) patients were overweight or obese,54% had central obesity and 12% had impaired glucose tolerance. Among the markers of atherosclerosis, hsCRP levels were nonsignificantly higher in patients with PCOS than in controls. The FMD was 12.18 ± 2.3% vs 8.3 ± 2.23% in patients with PCOS and controls respectively (P=0.01). CIMT was significantly different in two study groups (0.68 ± 0.11 in PCOS vs 0.52 ± 0.02 in normal subjects, (P=0.01). FMD had significant negative correlation with homeostasis model assessment (HOMA) index (r = -0.32, P=0.02) and hs CRP (r = -0.37, P=0.04) while hs CRP was correlated with BMI (r = 0.54, P=0.005), HOMA (r = 0.38, P=0.02) and FMD (r = -0.33, P=0.01). CIMT was significantly different in women with PCOS and control subjects, and it had significant correlation with age (r = 0.42, P=0.03), BMI (r = 0.36, P=0.01), waist circumference (r = 0.52, P=0.001) and HOMA (r = 0.31, P=0.04). Women with PCOS definitely have increased risk for future cardiovascular events. Clinicians should consider early cardiovascular screening and interventions to control all modifiable cardiovascular risk factors.


Normality and naturalness: a comparison of the meanings of concepts used within veterinary medicine and human medicine.
This article analyses the different connotations of "normality" and "being natural," bringing together the theoretical discussion from both human medicine and veterinary medicine. We show how the interpretations of the concepts in the different areas could be mutually fruitful. It appears that the conceptions of "natural" are more elaborate in veterinary medicine, and can be of value to human medicine. In particular they can nuance and correct conceptions of nature in human medicine that may be too idealistic. Correspondingly, the wide ranging conceptions of "normal" in human medicine may enrich conceptions in veterinary medicine, where the discussions seem to be sparse. We do not argue that conceptions from veterinary medicine should be used in human medicine and vice versa, but only that it could be done and that it may well be fruitful. Moreover, there are overlaps between some notions of normal and natural, and further conceptual analysis on this overlap is needed.


Energy homeostasis and cachexia in chronic kidney disease.
Loss of protein stores, presenting as clinical wasting, is reported to have a prevalence of 30-60% and is an important risk factor for mortality in chronic kidney disease (CKD) patients. There is debate as to whether the clinical wasting in CKD patients represents malnutrition or cachexia. Malnutrition results from inadequate intake of nutrients, despite a good appetite, and manifests as weight loss associated with adaptive metabolic responses such as decreased basic metabolic rate and preservation of lean body mass at the expense of fat mass. Furthermore, the abnormalities in malnutrition can usually be overcome simply by supplying more food or altering the composition of the diet. In contrast, cachexia is characterized by maladaptive responses such as anorexia, elevated basic metabolic rate, wasting of lean body tissue, and underutilization of fat tissue for energy. Diet supplementation and intradialytic parenteral nutrition have not been successful in reversing cachexia in CKD. The etiology of cachexia in CKD is complex and multifactorial. Two major factors causing muscle wasting in uremia are acidosis and decreased insulin responses. Inflammation secondary to cytokines may also play a significant role. The hypoalbuminemia of CKD patients is principally associated with inflammation and not changes in food intake. There is also recent evidence that hypothalamic neuropeptides may be important in the downstream signaling of cytokines in the pathogenesis of cachexia in CKD. Elevated circulating levels of cytokines, such as leptin, may be an important cause of uremia-associated cachexia via signaling through the central melanocortin system. Further research into the molecular pathways leading to cachexia may lead to novel therapeutic therapy for this devastating and potentially fatal complication of CKD.


Biochemical and genetic tests for inhibitors of Leishmania pteridine pathways.
The study of antifolate-resistant mutants of the protozoan parasite Leishmania has provided useful information about genetic processes such as gene amplification and mutation and knowledge of the unique features of the pteridine metabolic pathway in this primitive eukaryote. The novel bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) is an essential enzyme, yet most DHFR-TS inhibitors show little promise as potential drugs. Leishmania possess a novel alternative pteridine reductase (PTR1) which is relatively insensitive to methotrexate. We have proposed that the ability of PTR1 to serve as a metabolic bypass and thus modulate drug inhibition of DHFR-TS activity may be responsible for the poor efficacy of many antifolates. In this work, we have sought inhibitors of L. major PTR1 from a collection of 74 compounds. The most potent inhibitors were also tested against L. major DHFR-TS and human DHFR and several compounds showing good activity for PTR1 alone, or for all three reductases, were identified. The activity of these compounds was tested against wild-type promastigotes, and those which were potent inhibitors of both PTR1 and DHFR-TS (but not those active against only PTR1) showed good potencies. Growth inhibition tests of L. major mutants, lacking PTR1 or DHFR-TS (ptr1- and dhfr-ts- knockouts) or overexpressing PTR1, were used as a "genetic screen" to assess whether these two pteridine reductases were targets in vivo. Remarkably, only one compound showed a methotrexate-like pattern of inhibition. Six compounds showed good inhibition of Leishmania growth regardless of PTR1 or DHFR-TS levels. These findings suggest that Leishmania cells contain multiple targets for a diverse set of antifolates, with one or more significant targets in addition to DHFR-TS and PTR1. This emphasizes the necessity of combined biochemical and genetic screens in efforts to rationally design chemotherapeutic strategies in Leishmania.


Lower Success Rate of Débridement and Implant Retention in Late Acute versus Early Acute Periprosthetic Joint Infection Caused by Staphylococcus spp. Results from a Matched Cohort Study.
Surgical débridement, antibiotics and implant retention (DAIR) is currently recommended by international guidelines for both early acute (postsurgical) and late acute (hematogenous) periprosthetic joint infections (PJIs). However, due to a different pathogenesis of infection, a different treatment strategy may be needed. (1) Compared with early acute PJIs, are late acute PJIs associated with a higher risk of DAIR failure? (2) When stratified by microorganism, is the higher risk of failure in late acute PJI associated with Staphylocococcus aureus infection? (3) When analyzing patients with S. aureus infection, what factors are independently associated with DAIR failure? In this multicenter observational study, early acute and late acute PJIs treated with DAIR were retrospectively evaluated and matched according to treating center, year of diagnosis, and infection-causing microorganism. If multiple matches were available, the early acute PJI diagnosed closest to the late acute PJI was selected. A total of 132 pairs were included. Treatment success was defined as a retained implant during follow-up without the need for antibiotic suppressive therapy. Late acute PJIs had a lower treatment success (46% [60 of 132]) compared with early acute PJIs (76% [100 of 132]), OR 3.9 [95% CI 2.3 to 6.6]; p < 0.001), but the lower treatment success of late acute PJIs was only observed when caused by Staphylococcus spp (S. aureus: 34% versus 75%; p < 0.001; coagulase-negative staphylococci: 46% versus 88%; p = 0.013, respectively). On multivariable analysis, late acute PJI was the only independent factor associated with an unsuccessful DAIR when caused by S. aureus (OR 4.52 [95% CI 1.79 to 11.41]; p < 0.001). Although DAIR seems to be a successful therapeutic strategy in the management of early acute PJI, its use in late acute PJI should be reconsidered when caused by Staphylococcus spp. Our results advocate the importance of isolating the causative microorganism before surgery. Level III, therapeutic study.


Analysis by laser-induced breakdown spectroscopy of complex solids, liquids, and powders with an echelle spectrometer.
One of the most promising approaches to laser-induced breakdown spectroscopy (LIBS) experiments involves the use of an echelle spectrometer coupled with an intensified CCD. Even if drawbacks remain with its use, the echelle spectrometer facilitates a multielemental analysis that is more rapid than can be obtained with the more-conventional Czerny-Turner spectrometer and, moreover, does not sacrifice reliability. Quantitative results obtained with such apparatus for solids, liquids, powders, and gases are described and when possible compared with results from Czerny-Turner spectrometers. Liquid analysis by LIBS with echelle spectrometers has allowed a spectral database to be compiled. Once the qualitative spectra of pure elements in aqueous solutions, are obtained, they can be used for qualitative analysis of unknown samples.


[Ethical and methodological quality of non-interventional post-authorization studies promoted by Hospital Pharmacy Departments].
To describe the ethical and methodological quality of non-interventional post-authorization studies promoted by Hospital Pharmacy Departments (HPD). HPD promoted studies in the 2009-2011 period included in the Spanish Agency of Medicines and Medical Devices (AEMPS) registry and/or published in "Farmacia Hospitalaria" were identified. The most relevant ethical and methodological characteristics were analyzed. Studies promoted by HPD were also compared with studies not promoted by HPD. Twenty two studies promoted by HPD, and registered in the AEMPS were identified. Within the registered studies HPD promoted studies had lower sample size estimation (41,5% vs 80%) and international scope (0% vs 24%) compared to non HPD promoted studies with significant differences (p < 0,05). None of the published studies in the journal Farmacia Hospitalaria have been registered in the AEMPS and had lower methodological quality than the registered studies promoted by HPD in characteristics such as presence of control group (3,8% vs 27,3%) (p = 0,0072) and the sample size estimation of (19,2% vs 42,8%) (p < 0,05). The management and the methodological and ethical characteristics of the studies promoted by HPD should be improved according to the regulation. The registration in the AEMPS might have a positive impact on the quality of these research protocols.


An evaluation of supervised methods for identifying differentially methylated regions in Illumina methylation arrays.
Epigenome-wide association studies (EWASs) have become increasingly popular for studying DNA methylation (DNAm) variations in complex diseases. The Illumina methylation arrays provide an economical, high-throughput and comprehensive platform for measuring methylation status in EWASs. A number of software tools have been developed for identifying disease-associated differentially methylated regions (DMRs) in the epigenome. However, in practice, we found these tools typically had multiple parameter settings that needed to be specified and the performance of the software tools under different parameters was often unclear. To help users better understand and choose optimal parameter settings when using DNAm analysis tools, we conducted a comprehensive evaluation of 4 popular DMR analysis tools under 60 different parameter settings. In addition to evaluating power, precision, area under precision-recall curve, Matthews correlation coefficient, F1 score and type I error rate, we also compared several additional characteristics of the analysis results, including the size of the DMRs, overlap between the methods and execution time. The results showed that none of the software tools performed best under their default parameter settings, and power varied widely when parameters were changed. Overall, the precision of these software tools were good. In contrast, all methods lacked power when effect size was consistent but small. Across all simulation scenarios, comb-p consistently had the best sensitivity as well as good control of false-positive rate.


Heterogeneous reactions of alkylamines with ammonium sulfate and ammonium bisulfate.
The heterogeneous reactions between alkylamines and ammonium salts (ammonium sulfate and ammonium bisulfate) have been studied using a low-pressure fast flow reactor coupled to an ion drift-chemical ionization mass spectrometer (ID-CIMS) at 293 ± 2 K. The uptake of three alkylamines, i.e., monomethylamine, dimethylamine, and trimethylamine, on ammonium sulfate shows a displacement reaction of ammonium by aminium, evidenced by the release of ammonia monitored using protonated acetone dimer as the reagent ion. For the three alkylamines, the initial uptake coefficients (γ(0)) range from 2.6 × 10(-2) to 3.4 × 10(-2) and the steady-state uptake coefficients (γ(ss)) range from 6.0 × 10(-3) to 2.3 × 10(-4) and decrease as the number of methyl groups on the alkylamine increases. A different reaction mechanism is observed for the uptake of the three alkylamines on ammonium bisulfate, which is featured by an acid-base reaction (neutralization) with irreversible alkylamine loss and no ammonia generation and occurs at a rate limited by diffusion of gaseous alkylamines to the ammonium bisulfate surface. Our results reveal that the reactions between alkylamines and ammonium salts contribute to particle growth and alter the composition of ammonium sulfate and bisulfate aerosols in the atmosphere.


Amyloid Precursor Protein (APP) and GABAergic Neurotransmission.
The amyloid precursor protein (APP) is the parent polypeptide from which amyloid-beta (Aβ) peptides, key etiological agents of Alzheimer's disease (AD), are generated by sequential proteolytic processing involving β- and γ-secretases. APP mutations underlie familial, early-onset AD, and the involvement of APP in AD pathology has been extensively studied. However, APP has important physiological roles in the mammalian brain, particularly its modulation of synaptic functions and neuronal survival. Recent works have now shown that APP could directly modulate γ-aminobutyric acid (GABA) neurotransmission in two broad ways. Firstly, APP is shown to interact with and modulate the levels and activity of the neuron-specific Potassium-Chloride (K+-Cl-) cotransporter KCC2/SLC12A5. The latter is key to the maintenance of neuronal chloride (Cl-) levels and the GABA reversal potential (EGABA), and is therefore important for postsynaptic GABAergic inhibition through the ionotropic GABAA receptors. Secondly, APP binds to the sushi domain of metabotropic GABAB receptor 1a (GABABR1a). In this regard, APP complexes and is co-transported with GABAB receptor dimers bearing GABABR1a to the axonal presynaptic plasma membrane. On the other hand, secreted (s)APP generated by secretase cleavages could act as a GABABR1a-binding ligand that modulates presynaptic vesicle release. The discovery of these novel roles and activities of APP in GABAergic neurotransmission underlies the physiological importance of APP in postnatal brain function.


[Ureterohydronephrosis at pelvic prolapse in women].
The study was aimed to examination of prevalence of obstructive complications of the upper urinary tract in patients with pelvic prolapse and evaluation of the effectiveness of their correction using modern reconstructive surgery. Bilateral ureterohydronephrosis was detected in 14 (12%) of 117 patients. In patients with stage IV pelvic prolapse, proportion of patients with hydronephrosis was much higher--22.6%. Transvaginal reconstructive surgery with the installation of synthetic mesh prosthesis according to the Tension free Vaginal Mesh method ensured the restoration of normal anatomy of the pelvic floor and allowed to eliminate obstructive disorders of the upper urinary tract and their complications.


Sudden and unexpected death from pituitary tumor apoplexy.
Pituitary tumor apoplexy refers to a clinical syndrome precipitated by the expansion of a pituitary adenoma by hemorrhage or infarction. Individuals may present with myriad signs, including sudden onset of severe headache, visual changes, altered mental status, cranial nerve palsies, and hormonal dysfunction. This disorder constitutes a medical emergency and warrants an expedited evaluation, diagnosis, and treatment to prevent the potential sequelae of permanent visual loss, endocrine abnormalities, or death. We report a case of sudden death from undiagnosed pituitary tumor apoplexy. The decedent was evaluated by medical personnel on three occasions in the week prior to her death for severe headache, nausea, vomiting, and photophobia. Postmortem examination demonstrated a hemorrhagic infarction of a pituitary adenoma with necrosis and expansion out of the sella turcica. The recognition of and treatment for a patient with pituitary tumor apoplexy requires a rapid multidisciplinary effort. Failure of prompt diagnosis may be fatal and require a medico-legal death investigation for sudden and unexpected death.


[Adsorptive and catalytic properties of cholinesterase on phospholipid monolyers in different oxidation states].
Adsorption isotherm and enzymatic activity of protein interacting with the surface of solid carrier formed by oriented fat lipid chains of differently oxidized phospholipids have been studied. It has been found that the appearance of peroxide groups in fat acid chains results in a twofold decrease of protein limiting adsorption on the lipid monolayer. The data on enzymatic activity of proteins at variously filled suface with protein molecules indicate that the peroxide groups produce an activating effect under the conditions when protein interactions can be neglected. Superoxidation of phospholipid fat acids is suggested to be one of the mechanisms involved in the control of the processes proceeding on the membranes.


Secondary (AA) amyloidosis in cystic fibrosis. A report of three cases.
The authors report the pathologic features of three cases of amyloidosis associated with cystic fibrosis. Renal biopsy led to the diagnosis (case 1) or suspicion (case 2) of amyloidosis in patients who were 23 and 21 years old, respectively. The third patient died at age 22 years, and amyloidosis was not discovered until autopsy. Immunohistochemical staining and potassium-permanganate pretreatment of histologic sections in all three cases provided evidence that the amyloid seen in these patients is of the secondary (AA) type. Congo red staining in each case and electron microscopy in case 1 confirmed the initial diagnosis of amyloidosis. A markedly elevated serum amyloid A protein (160 micrograms/mL; normal less than 1 microgram/mL) in case 1 indicated the presence of large quantities of the precursor protein from which the AA fibrils of secondary amyloid are derived. The kidneys, spleen, and liver contained amyloid deposits in autopsy material from all three cases. Involvement of other organs by amyloid was variable. Review of autopsy material in Boston from 23 additional cystic fibrosis patients with long-term survival did not reveal any evidence of amyloidosis. It appears that secondary amyloidosis is emerging as a significant, although rare, complication of cystic fibrosis as greater numbers of these patients survive into adulthood.


Balloon sinuplasty: a minimally invasive option for patients with chronic rhinosinusitis.
Rhinosinusitis affects ~37 million people in the United States and accounts for almost 2% of all primary care office visits. Chronic rhinosinusitis (CRS) is often successfully managed in the primary care setting using antibiotics, topical or oral steroids, and saline nasal irrigation. Surgery is an option when medical management fails. Balloon sinuplasty is a minimally invasive endoscopic treatment, which is often used in combination with surgery. It aims to restore ostial patency with minimal mucosal damage, and it is indicated for dilatation of the paranasal sinuses for diagnostic and therapeutic purposes. The technology gently displaces, microfractures, and molds the bone surrounding the sinus outflow and may be used alone or in combination with conventional endoscopic surgery. Recent uncontrolled retrospective and prospective studies have reported the effectiveness and safety of balloon sinuplasty, including radiographic evidence of sinus patency and improved sinus-related quality of life scores for up to 2 years after balloon dilation. An examination of adverse events during a postmarketing assessment of balloon sinuplasty identified a total of 3 major complications among 28 500 patients, with a total of > 85 000 treated sinuses. While randomized controlled trials comparing balloon sinuplasty with conventional functional endoscopic sinus surgery have not been conducted, existing prospective and retrospective assessments suggest that balloon sinuplasty is a viable option for sinus intervention, either alone or in combination with conventional surgical treatment.


Fit Accuracy of Removable Partial Denture Frameworks Fabricated with CAD/CAM, Rapid Prototyping, and Conventional Techniques: A Systematic Review.
Analyzing and comparing the fit and accuracy of removable partial denture (RPDs) frameworks fabricated with CAD/CAM and rapid prototyping methods with conventional techniques. The present systematic review was carried out according to PRISMA guidelines. The search was carried out on PubMed/MEDLINE, Cochrane collaboration, Science direct, and Scopus scientific engines using selected MeSH keywords. The articles fulfilling the predefined selection criteria based on the fit and accuracy of removable partial denture (RPD) frameworks constructed from digital workflow (CAD/CAM; rapid prototyping) and conventional techniques were included. Nine full-text articles comprising 6 in vitro and 3 in vivo studies were included in this review. The digital RPDs were fabricated in all articles by CAD/CAM selective laser sintering and selective laser melting techniques. The articles that have used CAD/CAM and rapid prototyping technique demonstrated better fit and accuracy as compared to the RPDs fabricated through conventional techniques. The least gaps between the framework and cast (41.677 ± 15.546 μm) were found in RPDs constructed through digital CAD/CAM systems. A better accuracy was achieved using CAD/CAM and rapid prototyping techniques. The RPD frameworks fabricated by CAD/CAM and rapid prototyping techniques had clinically acceptable fit, superior precision, and better accuracy than conventionally fabricated RPD frameworks.


Adult ileo-ileo-caecal intussusception: case report and literature review.
Adult intussusception is a rare entity accounting for only 5% of all intussusceptions and causes approximately 1% of all adult intestinal obstructions. Unlike paediatric intussusceptions which are usually idiopathic, there is usually a lead point pathology which might be malignant in up to 50% cases. We present an unusual case of adult intussusception which was not diagnosed on any investigation including computerized tomographic (CT) scan and magnetic resonance imaging (MRI). It was a case of ileo-ileo-cecal intussusception caused by a large lipoma 38 mm × 43 mm × 61 mm. It was treated by emergency laparotomy for acute intestinal obstruction. A conservative resection with ileostomy was performed with good postoperative recovery.


Gender differences in diet-induced steatotic disease in Cyp2b-null mice.
Nonalcoholic fatty liver disease (NAFLD) is the most common liver disease; however, progression to nonalcoholic steatohepatitis (NASH) is associated with most adverse outcomes. CYP2B metabolizes multiple xeno- and endobiotics, and male Cyp2b-null mice are diet-induced obese (DIO) with increased NAFLD. However, the DIO study was not performed long enough to assess progression to NASH. Therefore, to assess the role of Cyp2b in fatty liver disease progression from NAFLD to NASH, we treated wildtype (WT) and Cyp2b-null mice with a normal diet (ND) or choline-deficient, L-amino acid-defined high fat diet (CDAHFD) for 8 weeks and determined metabolic and molecular changes. CDAHFD-fed WT female mice gained more weight and had greater liver and white adipose tissue mass than their Cyp2b-null counterparts; males experienced diet-induced weight loss regardless of genotype. Serum biomarkers of liver injury increased in both CDAHFD-fed female and male mice; however CDAHFD-fed Cyp2b-null females exhibited significantly lower serum ALT, AST, and ASP concentrations compared to WT mice, indicating Cyp2b-null females were protected from liver injury. In both genders, hierarchical clustering of RNA-seq data demonstrates several gene ontologies responded differently in CDAHFD-fed Cyp2b-null mice compared to WT mice (lipid metabolism > fibrosis > inflammation). Oil Red O staining and direct triglycerides measurements confirmed that CDAHFD-fed Cyp2b-null females were protected from NAFLD. CDAHFD-fed Cyp2b-null mice showed equivocal changes in fibrosis with transcriptomic and serum markers suggesting less inflammation due to glucocorticoid-mediated repression of immune responses. In contrast to females, CDAHFD-fed Cyp2b-null males had higher triglyceride levels. Results indicate that female Cyp2b-null mice are protected from NAFLD while male Cyp2b-null mice are more susceptible to NAFLD, with few significant changes in NASH development. This study confirms that increased NAFLD development does not necessarily lead to progressive NASH. Furthermore, it indicates a role for Cyp2b in fatty liver disease that differs based on gender.


Radiolabeling of [11C]FPS-ZM1, a receptor for advanced glycation end products-targeting positron emission tomography radiotracer, using a [11C]CO2-to-[11C]CO chemical conversion.
Aim: The receptor for advanced glycation end products (RAGE) is a viable target for early Alzheimer's disease (AD) diagnosis using positron emission tomography (PET) as RAGE overexpression precedes Aβ plaque formation. The development of a carbon-11 analog of FPS-ZM1 (N-benzyl-4-chloro-N-cyclohexylbenzamide, [11C]FPS-ZM1), possessing nanomolar affinity for RAGE, may enable the imaging of RAGE for early AD detection. Methodology & results: Herein we report an optimized [11C]CO2-to-[11C]CO chemical conversion for the synthesis of [11C]FPS-ZM1 and in vitro brain autoradiography. The [11C]CO2-to-[11C]CO conversion via 11C-silanecarboxylate derivatives was achieved with a 57% yield within 30 s from end of [11C]CO2 delivery. [11C]FPS-ZM1 was obtained with a decay-corrected isolated radiochemical yield of 9.5%. Conclusion: [11C]FPS-ZM1 distribution in brain tissues of wild-type versus transgenic AD model mice showed no statistically significant difference and high nondisplaceable binding.


Future needs and expected trends in peripheral blood cell analysis: erythrocyte histograms.
Erythrocyte histograms should be analysed in accordance with the ICSH recommended protocol to determine: How many red cell populations are present and the proportion of the total red cells which are included in each population. The central tendency (mode, median & mean) and dispersion (size ratio, geometric standard deviation & coefficient of variation) for each population. The proportion of microcytic or macrocytic cells in each population. Examples are given to show how the patient's diagnosis is assisted by careful analysis of the histograms and the effect of treatment monitored by sequential testing. Histograms can be generated from blood films or from aperture-impedance and light-scatter volume measurements. Volume measurements on these systems are influenced by the red cell shape and internal refractive index. Mean cell hemoglobin concentration affects both shape in the aperture-impedance orifice and internal refractive index in light-scatter systems. There is poor agreement between volume histograms obtained on the two measuring systems and the histograms available on current automated instruments provide no more useful information than can be obtained from the blood film. Automated instruments need to produce histograms with fewer artefacts and the histograms should then be examined in accordance with the ICSH protocol. This approach should maximise the diagnostic value of the complete blood count.


Intraspecific breakdown of self-incompatibility in Physalis acutifolia (Solanaceae).
Variation in mating systems is prevalent throughout angiosperms, with many transitions between outcrossing and selfing above and below the species level. This study documents a new case of an intraspecific breakdown of self-incompatibility in a wild relative of tomatillo, Physalis acutifolia. We used controlled greenhouse crosses to identify self-incompatible (SI) and self-compatible (SC) individuals grown from seed sampled across seven sites across Arizona and New Mexico. We measured 14 flower and fruit traits to test for trait variation associated with mating system. We also quantified pollen tube growth in vivo and tested for the presence of the S-RNase proteins in SI and SC styles. We found that seed from six of the seven sites produced SI individuals that terminated self-pollen tubes in the style and showed detectable S-RNase expression. By contrast, seed from one Arizona site produced SC individuals with no S-RNase expression. These SC individuals displayed typical selfing-syndrome traits such as smaller corollas, reduced stigma-anther distances, and a smaller pollen-ovule ratio. We also found plasticity in self-incompatibility as most of the SI individuals became SC and lost S-RNase expression roughly after 6 months in the greenhouse. While fixed differences in mating systems are known among the SI wild species and the often SC domesticated tomatillos, our study is the first to demonstrate intraspecific variation in natural populations as well as variation in SI over an individual's lifespan.


Effect of hysterosalpingography on serum cancer antigen (CA-125) levels.
The aim of our study was to detect any rise in serum levels of CA-125 after hysterosalpingography (HSG) as a result of irritation of the peritoneum. Serum levels of CA-125 were assessed before HSG and at the 2nd and 6th hours after HSG in 32 infertile patients who were admitted to our clinic from April 2002 to September 2002. Patients who showed tubal occlusion or Asherman Syndrome were excluded from the study. The levels of serum CA-125 were compared statistically. Statistical analysis was performed with SPSS 10.0 statistical software and the paired-sample t-test was applied. The mean levels of serum CA-125 before HSG, and at the 2nd and 6th hours after HSG were; 14.11 +/- 5.97 mIU/ml, 13.96 +/- 5.80 mIU/ml, and 14.06 +/- 5.80 mIU/ml, respectively. There was no statistically significant difference between serum levels of CA-125 before HSG and at the 2nd or 6th hours after HSG, and also between the 2nd and 6th hours after HSG (p > 0.05). We have not found any reports up to date about the levels of serum CA-125 after HSG. In our study we did not detect any increase in levels of serum CA-125 at the 2nd and 6th hours after HSG in spite of peritoneal irritation. We propose that this finding is because leakage of radioopaque material to the abdominal cavity is minimal during the HSG procedure.


Morphometry of the epidermis of an invasive megascoelecid earthworm (Amynthas gracilis, Kinberg 1867) inhabiting actively volcanic soils in the Azores archipelago.
For the first time, the structure, dimensions, and composition of the epidermis of an invasive earthworm species that has successfully colonized hostile conditions in actively volcanic soil on São Miguel (Azores) have been measured. Metal concentrations in actively volcanic (Furnas) and volcanically inactive (Fajã) soils were similar; however, Furnas soil was characterised by elevated temperature (10°C differential), relative hypoxia, extremely high CO(2) tension, and accompanying acidity. The epidermis of earthworm's resident at Fajã was approximately twice the thickness of the epidermis of conspecifics resident in Furnas soil. Reference worms transferred to Furnas soil for 14 days experienced an epidermal thinning of approximately 51%. In comparison, when Furnas earthworms were transferred to mesocosms at the relatively benign Fajã site, their epidermal thickness increased by approximately 21% over 14 days. Earthworms resident in Furnas soil had higher goblet cell counts than the residents of volcanically inactive soil on a neighbouring island (S. Maria). Transferring worms from S. Maria to mesocosms at Furnas induced a significant increase in goblet cell counts. Clearly, the active volcanic environment at Furnas poses a multifactorial stress challenge to the epigeic A. gracilis colonizer.


Brain structural correlates of irritability: Findings in a large healthy cohort.
Irritability and nonviolent aggression are common behavioral features across the population, yet there is limited neurobiological research into subclinical phenotypes representing the lower edge of a symptom continuum ranging from slight irritability to criminal violence. We studied brain structural correlates of irritability in a large healthy cohort to test the hypothesis of associations with fronto-limbic brain structures implicated in mood regulation. In a large multicenter effort, we recruited 409 mentally healthy adults from the community, who received T1-weighted high-resolution 3 T MRI scans. These structural scans were automatically preprocessed for voxel- and surface-based morphometry measurements with the CAT 12 toolbox implemented in SPM 12. Subclinical aggressive symptoms were assessed using the SCL-90-R aggression/hostility subscale and then correlated with cortical volume (VBM), and cortical thickness and gyrification. VBM analysis showed significant (P < 0.05, FDR-corrected at peak-level) positive correlations of cortical volume with SCL-90-R aggression subscale values in large clusters spanning bilateral anterior cingulate and orbitofrontal cortices and left lingual and postcentral gyri. Surface-based morphometry yielded mostly uncorrected positive correlations with cortical thickness in bilateral precentral gyri and with gyrification in left insula and superior temporal gyrus. Our findings imply an association of subclinical aggressive symptoms with cortical volume in areas important for emotion awareness and regulation, which might also be related to cortical adaptation to mental stress. These results overlap with several findings on impulsive aggression in patients suffering from affective and disruptive behavior disorders. They also suggest a biological symptom continuum manifesting in these brain areas. Hum Brain Mapp 38:6230-6238, 2017. © 2017 Wiley Periodicals, Inc.


[Changes in the indices of the atrial complex on a differentially amplified ECG in atrial infarct].
The atrial complex of differentially amplified ECG was examined in 19 patients with atrial infarction. The diagnosis of atrial infarction was based on the electrocardiographic features, and in 7 cases it was confirmed by postmortem examination. The electrocardiographic pattern was characterized by P-Q segment shift. Atrial infarction resulted in a decrease in the amplitude, the prolongation of atrial conduction time and the appearance of additional fragments in the atrial complex of differentially amplified ECG.


Cognitive-behavioural therapy and return-to-work intervention for patients on sick leave due to common mental disorders: a randomised controlled trial.
Common mental disorders (CMDs) cause great individual suffering and long-term sick leave. Cognitive-behavioural therapy (CBT) effectively treats CMDs, but sick leave is not reduced to the same extent as psychiatric symptoms. Research results regarding return-to-work interventions (RTW-Is) and their effect on sick leave are inconclusive. The aim of this study was to evaluate CBT, a RTW-I and combined CBT and RTW-I (COMBO) for primary care patients on sick leave due to CMDs. Patients with CMDs (n=211) were randomised to CBT (n=64), RTW-I (n=67) or COMBO (n=80). Sick-leave registry data after 1 year and blinded Clinician's Severity Rating (CSR) of symptoms post-treatment and at follow-ups after 6 and 12 months were primary outcomes. There was no significant difference between treatments in days on sick leave 1 year after treatment start (mean difference in sick-leave days range=9-27). CBT led to larger reduction of symptoms post-treatment (CSR; Cohen's d=0.4 (95% CI 0.1 to 0.8)) than RTW-I, whereas COMBO did not differ from CBT or RTW-I. At follow-up, after 1 year, there was no difference between groups. All treatments were associated with large pre-treatment to post-treatment improvements, and results were maintained at 1-year follow-up. No treatment was superior to the other regarding reducing sick leave. All treatments effectively reduced symptoms, CBT in a faster pace than RTW-I, but at 1-year follow-up, all groups had similar symptom levels. Further research is needed regarding how CBT and RTW-I can be combined more efficiently to produce a larger effect on sick leave while maintaining effective symptom reduction.


Comparative features of carcinoma in situ and atypical ductal hyperplasia of the breast on fine-needle aspiration biopsy specimens.
With the use of fine-needle aspiration biopsy to evaluate non-palpable breast lesions, an increasing number of cases of ductal carcinoma in situ (DCIS) are encountered. The authors previously demonstrated that it is not possible to distinguish definitively between DCIS and invasive ductal carcinoma on fine-needle aspiration biopsy. To determine whether DCIS could be separated from atypical ductal hyperplasia (ADH), the authors identified all cases of exclusive DCIS or ADH with fine-needle aspiration biopsy before surgery. Sixteen cases of ADH and 19 cases of DCIS were identified. Of these, 12 in each category were sufficiently cellular to allow evaluation of architectural and cytologic features. Cases of ADH were more likely to be diagnosed as negative or atypical (11 of 12); in contrast, DCIS was more likely to be designated as suspicious or positive (9 of 12). Architectural and cytologic features characteristic of ADH included cells arranged in flat cohesive sheets, distinct cell borders, and myoepithelial cells. Those features characteristic of DCIS were single cells representing more than 10% of atypical cells, cellular dyshesion, and inflammatory background, coarsely granular chromatin, and nuclear pleomorphism. Many other features were not useful in separating ADH from DCIS. Based on this small series, it appears that the distinction between some cases of DCIS and ADH may be possible on fine-needle aspiration biopsy.


Bioresorbable Bone Graft Composed of an RGD-Enriched Recombinant Human Collagen Polypeptide Induced Neovascularization and Regeneration of Mature Bone Tissue.
Bone graft materials provide a scaffold for migrating cells for bone regeneration. One of the major challenges is to support adequate neovascularization in the graft materials and bone tissue. Vascular endothelial cells have been shown to recognize the integrin-binding Arg-Gly-Asp (RGD) sequence in natural extracellular matrix (ECM) molecules. Here, we report a bone graft material composed of an RGD-enriched recombinant polypeptide based on human type I collagen alpha 1 chain (RCPhC1) and propose a category of bone graft materials called the recombinant bone matrix. RCPhC1 demonstrated significantly increased human umbilical vein endothelial cell attachment in vitro and was further processed through freeze casting and heat crosslinking processes to generate porous granular bone graft, in which RGD sequences remained canonical. When grafted in the rat model, RCPhC1 bone graft demonstrated a uniquely increased presence of CD34+ endothelial cells within the graft material. Bone tissue was found directly in contact with the pore structure of RCPhC1 bone graft, resulting in the regeneration of large bone tissue. By contrast, the combined demineralized and decellularized bone allograft containing bone collagen in the ECM did not show vascular formation within the graft material. When applied to canine tooth extraction socket, RCPhC1 bone graft rapidly induced highly vascularized regenerating tissues, which became a mature bone with the bone marrow tissue. These results indicate that RCPhC1 bone graft is a promising material and generated highly active bone tissues, which rapidly matured.


Measuring the physical cohesiveness of proteins using physical interaction enrichment.
Protein-protein interaction (PPI) networks are a valuable resource for the interpretation of genomics data. However, such networks have interaction enrichment biases for proteins that are often studied. These biases skew quantitative results from comparing PPI networks with genomics data. Here, we introduce an approach named physical interaction enrichment (PIE) to eliminate these biases. PIE employs a normalization that ensures equal node degree (edge) distribution of a test set and of the random networks it is compared with. It quantifies whether a set of proteins have more interactions between themselves than proteins in random networks, and can therewith be regarded as physically cohesive. Among other datasets, we applied PIE to genetic morbid disease (GMD) genes and to genes whose expression is induced upon infection with human-metapneumovirus (HMPV). Both sets contain proteins that are often studied and that have relatively many interactions in the PPI network. Although interactions between proteins of both sets are found to be overrepresented in PPI networks, the GMD proteins are not more likely to interact with each other than random proteins when this overrepresentation is taken into account. In contrast the HMPV-induced genes, representing a biologically more coherent set, encode proteins that do tend to interact with each other and can be used to predict new HMPV-induced genes. By handling biases in PPI networks, PIE can be a valuable tool to quantify the degree to which a set of genes are involved in the same biological process.


Polyethylene Terephthalate Textiles Enhance the Structural Maturation of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes.
Human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have the potential to serve as a model for human cardiomyocytes. However, hiPSC-CMs are still considered immature. CMs differentiated from hiPSCs more resemble fetal than adult cardiomyocytes. Putative factors enhancing maturation include in vitro culture duration, culture surface topography, and mechanical, chemical, and electrical stimulation. Stem cell-derived cardiomyocytes are traditionally cultured on glass surfaces coated with extracellular matrix derivatives such as gelatin. hiPSC-CMs are flat and round and their sarcomeres are randomly distributed and unorganized. Morphology can be enhanced by culturing cells on surfaces providing topographical cues to the cells. In this study, a textile based-culturing method used to enhance the maturation status of hiPSC-CMs is presented. Gelatin-coated polyethylene terephthalate (PET)-based textiles were used as the culturing surface for hiPSC-CMs and the effects of the textiles on the maturation status of the hiPSC-CMs were assessed. The hiPSC-CMs were characterized by analyzing their morphology, sarcomere organization, expression of cardiac specific genes, and calcium handling. We show that the topographical cues improve the structure of the hiPSC-CMs in vitro. Human iPSC-CMs grown on PET textiles demonstrated improved structural properties such as rod-shape structure and increased sarcomere orientation.


Lychee (Litchi chinensis Sonn.) Pulp Phenolic Extract Provides Protection against Alcoholic Liver Injury in Mice by Alleviating Intestinal Microbiota Dysbiosis, Intestinal Barrier Dysfunction, and Liver Inflammation.
Liver injury is the most common consequence of alcohol abuse, which is promoted by the inflammatory response triggered by gut-derived endotoxins produced as a consequence of intestinal microbiota dysbiosis and barrier dysfunction. The aim of this study was to investigate whether modulation of intestinal microbiota and barrier function, and liver inflammation contributes to the hepatoprotective effect of lychee pulp phenolic extract (LPPE) in alcohol-fed mice. Mice were treated with an ethanol-containing liquid diet alone or in combination with LPPE for 8 weeks. LPPE supplementation alleviated ethanol-induced liver injury and downregulated key markers of inflammation. Moreover, LPPE supplementation reversed the ethanol-induced alteration of intestinal microbiota composition and increased the expression of intestinal tight junction proteins, mucus protecting proteins, and antimicrobial proteins. Furthermore, in addition to decreasing serum endotoxin level, LPPE supplementation suppressed CD14 and toll-like receptor 4 expression, and repressed the activation of nuclear factor-κB p65 in the liver. These data suggest that intestinal microbiota dysbiosis, intestinal barrier dysfunction, and liver inflammation are improved by LPPE, and therefore, the intake of LPPE or Litchi pulp may be an effective strategy to alleviate the susceptibility to alcohol-induced hepatic diseases.


Comparison of six ECMO selection criteria and analysis of factors influencing their accuracy.
This study compared six extracorporeal membrane oxygenation (ECMO) selection criteria in 42 neonates and analyzed factors influencing the accuracy of outcome predictions. The sensitivity of the criteria in identifying fatal cases varied from 0.44 to 0.94 and the specificity of predictions of survival ranged from 0.42 to 0.69. The criterion having the highest sensitivity had the lowest specificity and conversely the criterion with the lowest sensitivity had the highest specificity. Overall accuracy of the criteria, as measured by the total number of correct outcome predictions, differed little among the criteria (23/42 to 27/42 correct predictions). Three factors influenced predictive accuracy: 1) a primary diagnosis of congenital diaphragmatic hernia (CDH) was associated with a greater mortality (P less than 0.001) and a significantly higher positive predictive value (PPV) for all criteria (P = 0.0009-0.012) than that seen in patients with other primary diagnoses; 2) calculating the alveolar-arterial oxygen gradient using an assumed, rather than measured barometric pressure, or estimating oxygenation index using a calculated, rather than a measured, mean airway pressure, increased false positive mortality predictions in non-CDH patients; and 3) requiring a peak inspiratory pressure (PIP) of at least 50 cm H2O in the definition of maximal medical management, rather than a PIP of 20-49 cm H2O, significantly increased the PPV for three of four criteria examined (P = 0.02-0.04). Awareness of these factors may facilitate the identification of neonates who need ECMO to survive.


Phylogenomic analyses reveal a deep history of hybridization and polyploidy in the Neotropical genus Lachemilla (Rosaceae).
Hybridization, incomplete lineage sorting, and phylogenetic error produce similar incongruence patterns, representing a great challenge for phylogenetic reconstruction. Here, we use sequence capture data and multiple species tree and species network approaches to resolve the backbone phylogeny of the Neotropical genus Lachemilla, while distinguishing among sources of incongruence. We used 396 nuclear loci and nearly complete plastome sequences from 27 species to clarify the relationships among the major groups of Lachemilla, and explored multiple sources of conflict between gene trees and species trees inferred with a plurality of approaches. All phylogenetic methods recovered the four major groups previously proposed for Lachemilla, but species tree methods recovered different topologies for relationships between these four clades. Species network analyses revealed that one major clade, Orbiculate, is likely of ancient hybrid origin, representing one of the main sources of incongruence among the species trees. Additionally, we found evidence for a potential whole genome duplication event shared by Lachemilla and allied genera. Lachemilla shows clear evidence of ancient and recent hybridization throughout the evolutionary history of the group. Also, we show the necessity to use phylogenetic network approaches that can simultaneously accommodate incomplete lineage sorting and gene flow when studying groups that show patterns of reticulation.


Perceptions of "futile care" among caregivers in intensive care units.
Many caregivers in intensive care units (ICUs) feel that they sometimes provide inappropriate or excessive care, but little is known about their definition of "futile care" or how they attempt to limit its impact. We sought to explore how ICU staff define medically futile care, why they provide it and what strategies might promote a more effective use of ICU resources. Using semi-structured interviews, we surveyed 14 physician directors, 16 nurse managers and 14 respiratory therapists from 16 ICUs across Ontario. We analyzed the transcripts using a modified grounded-theory approach. From the interviews, we generated a working definition of medically futile care to mean the use of considerable resources without a reasonable hope that the patient would recover to a state of relative independence or be interactive with his or her environment. Respondents felt that futile care was provided because of family demands, a lack of timely or skilled communication, or a lack of consensus among the treating team. Respondents said they were able to resolve cases of futile care most effectively by improving communication and by allowing time for families to accept the reality of the situation. Respondents felt that further efforts to limit futile care should focus on educating the public and health care professionals about the role of the ICU and about alternatives such as palliative care; mandating early and skilled discussion of resuscitation status; establishing guidelines for admission to the ICU; and providing legal and ethical support for physicians who encounter difficulties. There was a broad consistency in responses among all disciplines. ICU physicians, nurses and respiratory therapists have similar and well-formed opinions about how to define and resolve medically futile care and where to focus future efforts to limit the impact of futile care in the ICU.


An inexpensive, efficient method for regular egg collection from zebrafish in a recirculating system.
Zebrafish in our laboratory are usually bred by removing the fish from the recirculating aquatic system and placing them into 1-2 L spawning tanks. These spawning tanks consist of a bottom reservoir, a lid, and an insert that fits in closely into the bottom reservoir. When the fish breed, the eggs fall through holes of the insert and into the reservoir, thus preventing them from being cannibalized. Because fish in these spawning tanks are not fed and do not get fresh water, they are bred only once a week. During a period where we had high demand for embryos, we instead tried breeding the fish for multiple consecutive days on the recirculating system. Fish were placed into the spawning insert as usual, but the insert was placed into the home tank instead of into the bottom reservoir. We found that there was no significant difference in the number of fertilized eggs produced between the spawning tank and home tank breeding methods. Further, the fish in the home tanks regularly produced fertile embryos over a 28-day time course, with the highest number of eggs per pair produced by the tank with only one pair of adult fish. This method is time-saving as fish bred in home tanks only require to be set up once. It is also an effective way to collect embryos over long periods from the same pair or group of fish and to more easily obtain embryos from stocks with low spawning frequency.


Prognostic value of DNA ploidy using flow cytometry in 1301 breast cancer patients: results of the prospective Multicenter Morphometric Mammary Carcinoma Project.
The literature on breast cancer reports conflicting prognostic results with respect to DNA ploidy of flow cytometric DNA histograms. This might result from different DNA ploidy classification methods. Our study evaluated the prognostic power of DNA ploidy, using different classification methods, in a large prospective group (n = 1301) of breast cancer patients. Flow cytometric DNA histograms obtained from fresh frozen material were interpreted with use of a commercially available computer program. On the basis of the number of stemlines and the DNA Index, we classified the DNA ploidy by different methods. In all of the cases, the classification method "DNA diploid versus DNA nondiploid" provided the best prognostic significance for overall survival (OS) (Mantel-Cox (MC) = 5.4, P = .02; relative risk (RR) = 1.3, P = .05) and for disease-free survival (DFS) (MC = 11.8, P = .0006; RR = 1.3, P < .05). This was also true for the OS of the lymph node-positive (but not the lymph node-negative) subgroup (MC = 4.1, P = .04; RR = 1.3, P = .05). In subgroups classified on the basis of tumor size, DNA ploidy showed prognostic significance for DFS only in the subgroup of tumors smaller than 2 cm and larger than 5 cm. In multivariate analysis, DNA ploidy showed no additional prognostic power to lymph node status and tumor size. The classification "DNA diploid versus DNA nondiploid" was mostly consistent with respect to prognostic power for OS and DFS, especially in small or lymph node-positive tumors. The RR of DNA nondiploid patients was only marginally higher, however, so large study groups are required to reach statistical significance. This could partly explain the disagreements in the literature. Therefore, DNA ploidy seems to be of little clinical importance in breast cancer patients, compared with other prognostic parameters.


Graviresponsiveness and abscisic-acid content of roots of carotenoid-deficient mutants of Zea mays L.
The abscisic-acid (ABA) content of roots of the carotenoid-deficient w-3, vp-5, and vp-7 mutants of Z. mays was analyzed using gas chromatography-mass spectrometry with an analysis sensitivity of 6 ng ABA g(-1) fresh weight (FW). Roots of normal seedlings of the same lines were characterized by the following amounts of ABA (as ng ABA g(-1) FW,±standard deviation): w-3, 279±43; vp-5, 237±26; vp-7, 338±61. We did not detect any ABA in roots of any of the mutants. Thus, the lack of carotenoids in these mutants correlated positively with the apparent absence of ABA. Primary roots of normal and mutant seedlings were positively gravitropic, with no significant differences in the curvatures of roots of normal as compared with mutant seedlings. These results indicate that ABA 1) is synthesized in maize roots via the carotenoid pathway, and 2) is not necesary for positive gravitropism by primary roots of Z. mays.


Synthesis of multi-functional large pore mesoporous silica nanoparticles as gene carriers.
The development of functional nanocarriers that can enhance the cellular delivery of a variety of nucleic acid agents is important in many biomedical applications such as siRNA therapy. We report the synthesis of large pore mesoporous silica nanoparticles (LPMSN) loaded with iron oxide and covalently modified by polyethyleneimine (denoted PEI-Fe-LPMSN) as carriers for gene delivery. The LPMSN have a particle size of ∼200 nm and a large pore size of 11 nm. The large pore size is essential for the formation of large iron oxide nanoparticles to increase the magnetic properties and the adsorption capacity of siRNA molecules. The magnetic property facilitates the cellular uptake of nanocarriers under an external magnetic field. PEI is covalently grafted on the silica surface to enhance the nanocarriers' affinity against siRNA molecules and to improve gene silencing performance. The PEI-Fe-LPMSN delivered siRNA-PLK1 effectively into osteosarcoma cancer cells, leading to cell viability inhibition of 80%, higher compared to the 50% reduction when the same dose of siRNA was delivered by a commercial product, oligofectamine.


Protective mechanisms of tea polyphenols regulating the PI3K/Akt pathway on early brain injury after subarachnoid hemorrhage in rats.
In recent years, numerous studies have demonstrated that tea polyphenols (TPPs) can exert neuroprotective effects through the regulation of the PI3K/Akt pathway. The objective of this work was to verify whether TPPs could protect against early brain injury in rats after subarachnoid hemorrhage (SAH) by modulating the PI3K/Akt pathway. A total of 150 rats were randomly rolled into control (C), TPP, and SAH groups. The TPP and SAH groups underwent endovascular perforation to induce SAH, while C group received only endovascular needle puncture and saline injection. Brain water content, Evans Blue (EB) extravasation assay, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining, Western blot, and RT-PCR analyses were performed. Relative to SAH group, TPP treatment considerably improved neurological function scores following SAH, reduced brain edema, cortical neuronal apoptosis, and blood-brain barrier damage. Levels of aquaporin-4 (AQP4) and apoptosis-related protein Bax were considerably lower in the TPP group than in SAH group. Conversely, levels of anti-apoptotic protein Bcl-2 and tight junction protein Zona occludens 1 (ZO-1) were considerably higher in the TPP group. Furthermore, TPP treatment was found to activate the PI3K/Akt signaling. TPPs can mitigate early brain injury caused by SAH in rats by reducing AQP4 levels, alleviating cortical damage, and attenuating neuronal apoptosis. These findings elucidate the protective mechanisms of TPPs against early brain injury following SAH through the regulation of the PI3K/Akt signaling.


Protective roles of meditation practice and self-esteem on cognitive functions over time: findings from the Midlife in the United States study.
Cognitive function is an important component of healthy ageing. However, it is unclear whether relaxation/meditation practices provide cognitive benefits, particularly in midlife and early late life. Meditative practices are associated with higher self-esteem, and self-esteem serves as a general protective factor for many health outcomes. The current study examines associations between meditation practice, self-esteem, and change over 10 years in midlife cognitive performance. Data were from waves 2 (2003-2004) and 3 (2013-2014) of the Midlife in the United States (MIDUS) study. We used structural equation modelling to examine whether persistent meditation practice at both waves or episodic practice at one wave is associated with better cognitive function over 10 years, compared to no meditation practice, while controlling for prior cognitive function and covariates (baseline socio-demographics, health, and functional status). Additionally, we assessed if self-esteem mediates the above associations. We included 2168 individuals (Mage = 65 ± 11). After controlling for covariates, the findings revealed that persistent meditation practice in both waves was associated with significantly less decline in episodic memory; however, no such effects were found for executive function. Further, although participants' higher self-esteem was significantly associated with less decline in executive function and episodic memory, it did not mediate the associations between meditation practice and cognitive functions. While both persistent meditation practice and self-esteem have associations with cognitive outcomes for middle-aged and older adults in MIDUS, self-esteem as a mediator was not supported. Thus, future investigations should examine mechanisms that underlie these protective factors on cognitive performances across adulthood.


The East coast magnetic anomaly.
A model is proposed for the East Coast Magnetic Anomaly that, for the first time, incorporates an observed basement feature-a normal fault that consistently lies below the landward flank of the anomaly. The East Coast Magnetic Anomaly is similar to slope anomalies found over passive continental margins in many parts of the world. Thus the proposed model can be used to interpret slope anomalies in general.


Effects of toxic agents at the protein level: quantitative measurement of 213 mouse liver proteins following xenobiotic treatment.
By analyzing two-dimensional electrophoretic patterns of mouse liver proteins with a computerized image analysis system, we have observed quantitative changes in the abundance of more than 70 proteins in mice treated with various agents. Aroclor 1254, a mixture of polychlorinated biphenyls known to induce a broad spectrum of microsomal activity, induces the largest group of changes (60 proteins altered at p less than 0.001 significance). Phenobarbital produces a small set of characteristic changes that forms part of the much larger Aroclor 1254 effect. Ibuprofen treatment produces a phenobarbital-like pattern of change, with the addition of at least one protein change not observed with any of the other treatments. Cycloheximide and carbon tetrachloride each induces a different characteristic pattern of protein alteration. We have assigned most of the mouse liver proteins to a specific subcellular fraction, and it appears that the predominant class of proteins altered by each compound is present in the soluble phase, rather than in the microsomal fraction. The ability to survey large numbers of tissue proteins for involvement in pharmacologic and toxic effects may allow a more comprehensive understanding of the mechanisms of action in vivo and provide new markers of tissue damage.


More than inorganic copper is bioavailable to aquatic mosses at environmentally relevant concentrations.
The present study investigates how dissolved organic matter (DOM) alters copper bioavailability at environmentally relevant concentrations (1-5 microg/L of dissolved copper, 1-4 mg/L of dissolved organic copper). A methodology combining two biological endpoints (short-term and steady-state bioaccumulation of copper by the aquatic moss Fontinalis antipyretica) and a sampling of labile copper with diffusion gradient in thin films (DGT) is proposed for batch experiments conducted with mineral water and various DOM, ethylenediaminetetra-acetic acid (EDTA), humic acid, and natural Seine River (France) extracts (hydrophobic and transphilic fractions). All types of DOM reduce the bioavailability of copper to aquatic mosses, and this reduction was more pronounced for the short-term biological endpoint, which was taken as being representative for environmental exposure. Labile copper sampled with DGT made it possible to estimate short-term bioaccumulation in the case of EDTA and natural Seine River extracts. With humic acid solutions, however, labile copper was lower than bioavailable copper. This result suggests that at realistic metal concentrations and with certain types of natural DOM, bioavailable copper might comprise not only inorganic copper but also some weak organic complexes. Hence, labile copper, in situ sampled with DGT, might not systematically overestimate bioavailable copper, as suggested previously on the basis of in vitro toxicity studies.


Agents which increase cyclic AMP have diverse effects on low-density-lipoprotein-receptor function in human vascular smooth-muscle cells and skin fibroblasts.
Receptor-mediated binding and metabolism of low-density lipoproteins (LDL) in cultured human vascular smooth-muscle cells and skin fibroblasts are altered by increased cellular cyclic AMP concentrations. However, the LDL receptor does not respond to changes in cyclic AMP concentration in a simple manner. The activation of adenylate cyclase with forskolin, or the addition of membrane-permeant cyclic AMP analogues, initially decreases the expression of the LDL receptor, but is followed by a substantial increase in receptor expression after 24 h. This increase does not occur in the presence of inhibitors of RNA or protein synthesis, and is due to doubling of the Bmax. of the LDL receptor, without alteration of its affinity for LDL. By contrast, elevation of cyclic AMP concentration by inhibition of phosphodiesterases results in decreased receptor expression throughout the 24 h period. These two response patterns are reproducible phenomena, consistently observed in low-passaged cells derived from seven unrelated individuals.


Proteins and protein fragments in nephrotic syndrome: Clusters, specificity and mechanisms.
Proteinuria is the hallmark of renal diseases and the characterization of the urinary protein composition may become an important source of information for diagnosis and research. So far, protein analysis in urine has been utilized for a generic individuation of site-specific defects (glomerular vs. tubular) but there is a need for an extension of proteomics to specific urinary biomarkers in selected clinical conditions. The identification of fragments of proteins in plasma and urine may increase the spectrum of urinary biomarkers. The unique speculative application so far proposed for protein fragments is nephrotic syndrome, and specifically focal segmental glomerulosclerosis, in which case they reflect intrinsic proteolysis occurring in plasma and represent surrogate biomarkers of the disease activity. Albumin is probably the most studied protein. Several of the albumin fragments present a peculiar distribution of the fingerprint peptide pattern containing both the N-terminal region and the C-terminal domain with a complete lack of any MS signals for the internal sequence region. Their characterization utilizing new strategies based on 2-D nondenaturing electrophoresis is now in progress. Studies on a direct characterization of proteases in plasma and urine will also define the participation of proteases to the genesis of renal diseases.


Alterations in substance P binding in brain nuclei of spontaneously hypertensive rats.
Substance P binding sites were characterized in brain nuclei of young (4-wk-old) and adult (16-wk-old) spontaneously hypertensive rats (SHR) and age-matched normotensive Wistar-Kyoto (WKY) control rats by quantitative autoradiography. Young SHR presented higher affinity constants (KA) than young WKY. The changes were restricted to the locus coeruleus, the area postrema, the dorsal motor nucleus of the vagus, and to discrete areas located in lobes 9 and 10 of the vermis cerebelli of SHR. There were no differences in the maximal binding capacity (Bmax) except in the nucleus ambiguus where the Bmax was lower than in WKY. Conversely, the number of substance P binding sites was higher in the locus coeruleus, the nucleus tegmentalis dorsalis, the nucleus ambiguus, the dorsal motor nucleus of the vagus, the hypoglossal nucleus, the inferior olivary nucleus, and lobes 9 and 10 of the vermis cerebelli of adult SHR when compared with adult WKY. Our results support the hypothesis of a role for brain substance P in blood pressure regulation and in genetic hypertension in rats.


Effects of load on good morning kinematics and EMG activity.
Many strength and conditioning coaches utilize the good morning (GM) to strengthen the hamstrings and spinal erectors. However, little research exists on its electromyography (EMG) activity and kinematics, and how these variables change as a function of load. The purpose of this investigation was to examine how estimated hamstring length, integrated EMG (IEMG) activity of the hamstrings and spinal erectors, and kinematics of the lumbar spine, hip, knee, and ankle are affected by changes in load. Fifteen trained male participants (age = 24.6 ± 5.3 years; body mass = 84.7 ± 11.3 kg; height = 180.9 ± 6.8 cm) were recruited for this study. Participants performed five sets of the GM, utilizing 50, 60, 70, 80, and 90% of one-repetition maximum (1RM) in a randomized fashion. IEMG activity of hamstrings and spinal erectors tended to increase with load. Knee flexion increased with load on all trials. Estimated hamstring length decreased with load. However, lumbar flexion, hip flexion, and plantar flexion experienced no remarkable changes between trials. These data provide insight as to how changing the load of the GM affects EMG activity, kinematic variables, and estimated hamstring length. Implications for hamstring injury prevention are discussed. More research is needed for further insight as to how load affects EMG activity and kinematics of other exercises.


Mastering mouse lung transplantation from scratch--a track record.
Mouse lung transplantation has evolved into an established scientific model that is currently used by an increasing number of research groups. Acquiring this technique without previous microsurgical knowledge is considered very difficult. Disclosing all the intraoperative failures and mistakes during the model's evolution will encourage all researchers who lack microsurgical skills that overcoming and eventually succeeding in this model is possible. Inbred (C57BL/6, BALB/c, SVG129) and CD1-outbred mice served as the transplant donors and recipients. The training procedure was performed by a surgeon not experienced in microsurgery, and arranged as follows: donor preparation until proof of functionality, graft implantation into deceased recipients, and graft implantation into surviving recipients until stable performance was achieved. The transplant's viability was controlled using micro-computed tomography imaging. Donor preparation complications decreased from 43% after 1 month to 0% after 2 mo. The first functional donor was implanted at day 28, and the first successful implantation into a surviving recipient was performed at day 60 after six training recipients. Micro-computed tomography confirmed a ventilated and perfused graft. Intraoperative complications, mainly due to anastomosis failure, decreased from 58% after the first month to 15% at the latest assessment. The most recent implantation time was 75 ± 4.8 min, and the transplantation success rate was 82% ± 2.8%. A modified forceps considerably improved completion of the venous anastomosis. Consistent success in the mouse lung transplantation model can be achieved even without pre-existing microsurgical skills. The surgery can be mastered within a reasonable period using a limited number of training animals. Procedure-related complications can be restricted to a minimum by applying key corrective steps at critical phases. This should encourage investigators without pre-expert knowledge in microsurgery to start to learn this research model.


Molecular and functional characterization of inositol trisphosphate receptors during early zebrafish development.
Fluctuations in cytosolic Ca(2+) are crucial for a variety of cellular processes including many aspects of development. Mobilization of intracellular Ca(2+) stores via the production of inositol trisphosphate (IP(3)) and the consequent activation of IP(3)-sensitive Ca(2+) channels is a ubiquitous means by which diverse stimuli mediate their cellular effects. Although IP(3) receptors have been well studied at fertilization, information regarding their possible involvement during subsequent development is scant. In the present study we examined the role of IP(3) receptors in early development of the zebrafish. We report the first molecular analysis of zebrafish IP(3) receptors which indicates that, like mammals, the zebrafish genome contains three distinct IP(3) receptor genes. mRNA for all isoforms was detectable at differing levels by the 64 cell stage, and IP(3)-induced Ca(2+) transients could be readily generated (by flash photolysis) in a controlled fashion throughout the cleavage period in vivo. Furthermore, we show that early blastula formation was disrupted by pharmacological blockade of IP(3) receptors or phospholipase C, by molecular inhibition of the former by injection of IRBIT (IP(3) receptor-binding protein released with IP(3)) and by depletion of thapsigargin-sensitive Ca(2+) stores after completion of the second cell cycle. Inhibition of Ca(2+) entry or ryanodine receptors, however, had little effect. Our work defines the importance of IP(3) receptors during early development of a genetically and optically tractable model vertebrate organism.


Lymphocytic choriomeningitis virus. III. Structural proteins of the virion.
Analysis of radioactively labelled and highly purified infectious lymphocytic choriomeningitis (LCM) virus by polyacrylamide gel electrophoresis (PAGE) revealed 12 components which, according to their apparent molecular weight and glycosylation status, were designated as p19, p25, p26, gp35, p38, gp44, gp60, p63, p77, gp85, gp130, and p200. As shown by immunoprecipitation, they all bound to rabbit anti-LCM virus antibodies. Three proteins, namely gp35 (= 'GP-2'), gp44 (= 'GP-1') and p63 (= 'NP'), had previously been described by others as major constituents of the virion. Our results confirm this and suggest that gp60, p77, gp85, and p200 are further distinct structural proteins. In contrast, p25 and p38 appear to be cleavage or degradation products of p63; p19 and p26 seem to belong to gp60, which could be the monomeric form of a dimer, gp130. Peptide mapping by limited proteolysis revealed considerable overlapping of amino acid sequences among the major glycoproteins with one peptide being common to all. From the results of PAGE performed after external labelling of intact virions, we conclude that gp44, gp60, and gp85 (but not gp35) form the surface of the virus envelope. Analytical isoelectric focusing under non-reducing conditions has shown that the major glycoproteins appeared to consist of several components with different isoelectric points.


Considerations for a better definition of male orgasmic disorder in DSM V.
All of the Diagnostic and Statistical Manual of Mental Disorders, 4th Ed., text revision (DSM-IV-TR) criteria for sexual disorders have been criticized on multiple grounds, including that the criteria lack precision, that the requirement of marked distress is inappropriate, and that the specification of etiological subtypes should be deleted. The goal of this article is to review evidence relevant to diagnostic criteria for male orgasmic disorder published since 1990. Medline searches from 1990 forward were conducted using the terms male orgasmic disorder, anorgasmia, delayed ejaculation, retarded ejaculation, ejaculatory delay, and ejaculatory disorder. Early drafts of proposed alterations in diagnostic criteria were submitted to advisors. Evidence reviewed was judged by current usage of terminology, evidence allowing precise definition of the syndrome, and evidence concerning separation of the syndrome from distress. The literature search indicated minimal use of the term male orgasmic disorder and minimal knowledge concerning psychogenic ejaculatory problems. It is recommended that the term male orgasmic disorder be replaced with the term delayed ejaculation. Duration and severity criteria are recommended. Since many ejaculatory problems are idiopathic, it is recommended that the etiological subtypes due to psychological or due to combined factors be eliminated.


Protective effects of trifluoperazine on the microcirculation of cold-stored livers.
Previous studies have shown a protective effect of trifluoperazine (TFP), a calmodulin inhibitor, upon the microcirculation of cold-stored kidneys. The present study points to similar beneficial effects of TFP on the microcirculation of cold-stored livers; 25 canine livers were preserved for 24 hr with Euro-Collins' solution (EC) (n = 8), University of Wisconsin solution (UW) (n = 7), or UW + TFP (n = 10). The stored livers underwent heterotopic transplantation (HLTX); hepatic-artery and portal-vein pressure and flow were monitored; oxygen consumption and extraction were measured before HLTX and at 15-min intervals after reperfusion, for 1 hr. Mean hepatic-artery and portal-vein flow (HAF & PVF) prior to donor hepatectomy were 172 and 530 cc/min, respectively. Poor HAF and PVF occurred in EC-HLTX (mean 35, 175 cc/min, respectively). The damaged EC-flushed livers could not compensate to the decreased hepatic blood flow by increased oxygen extraction (oxygen consumption and extraction, 8.7 vol.% and 48%, respectively). Light and electron microscopy showed severe liver necrosis and periportal hemorrhages. Improved hepatic-artery and portal-vein flows were seen in UW HLTX (105 and 254 cc/min), and oxygen consumption and extraction were 16.4 vol.% and 66%, respectively. Liver biopsy taken just before reperfusion revealed well-preserved liver architecture. Liver biopsy obtained 1 hr after reperfusion revealed marked edema of the portal triad, sinusoid congestion, and hemorrhage. Electron-microscopy biopsies obtained during reperfusion at 15-min intervals revealed severe vasospasm of the terminal hepatic arterioles and progressive damage to the liver microcirculation. The addition of TFP to the UW-flush solution resulted in excellent protection of the liver microcirculation. Marked increase in hepatic-artery and portal-vein blood flow was noted after reperfusion (mean 167 and 421 cc/min, respectively (P 0.02 vs. UW: P 0.001 vs. EC). The recovery of metabolic activity was evident by the high oxygen consumption and extraction (25.8 vol.% and 80%, respectively). And serial liver biopsies obtained after reperfusion have shown excellent protection of liver architecture and the absence of hepatic arteriolar vasospasm. Taken together, these data suggest that the addition of TFP to the UW solution protects the liver microcirculation by rendering the hepatic microcirculation insensitive to vasospastic stimuli during reperfusion, thus permitting better metabolic recovery after transplantation.


Water, water everywhere: environmental DNA can unlock population structure in elusive marine species.
Determining management units for natural populations is critical for effective conservation and management. However, collecting the requisite tissue samples for population genetic analyses remains the primary limiting factor for a number of marine species. The harbour porpoise (Phocoena phocoena), one of the smallest cetaceans in the Northern Hemisphere, is a primary example. These elusive, highly mobile small animals confound traditional approaches of collecting tissue samples for genetic analyses, yet their nearshore habitat makes them highly vulnerable to fisheries by-catch and the effects of habitat degradation. By exploiting the naturally shed cellular material in seawater and the power of next-generation sequencing, we develop a novel approach for generating population-specific mitochondrial sequence data from environmental DNA (eDNA) using surface seawater samples. Indications of significant genetic differentiation within a currently recognized management stock highlights the need for dedicated eDNA sampling throughout the population's range in southeast Alaska. This indirect sampling tactic for characterizing stock structure of small and endangered marine mammals has the potential to revolutionize population assessment for otherwise inaccessible marine taxa.


[Diagnosis of neurometabolic and neurodegenerative diseases by cutaneous biopsy].
Cutaneous biopsy is of great use in the diagnosis of neurodegenerative diseases, especially neurometabolic and inborn errors of metabolism. The sweat glands, especially the eccrine ones, and certain cellular elements in the adventitial dermis like the Schwann cells, axons, endothelium and fibroblasts, are altered in inborn errors of metabolism and in certain neurodegenerative diseases. The morphology and placement of these deposits, as for example axonal spheroids, Lafora bodies, homogeneous cytoplasmic bodies and prismatic bodies, permit to diagnosis the disease. Differential diagnosis must be made with simulating storage disorders. The study of skin biopsy must be complete and rigorous and follow a system of work, that includes electron microscopy, enzymatic determination and, occasionally, molecular biology. Valuable information about neurodegenerative diseases as muscular dystrophies, axonal neuropathy, Alzheimer disease and others can be carried out by skin biopsy. It can be foreseen that in the future indications of skin biopsy will be extended to other neurodegenerative diseases.


[Nursing in the movies: its image during the Spanish Civil War].
The cinema had carried out a determining role in the development of stereotypes and in a wide gamut of models related to real life situations. The objective of this analysis is to determine the influence cinema had on the image of nurses during the Spanish Civil War from 1936-1939. These are the initial hypotheses: the role of Spanish nurses during the civil war was reflected by both sides in their respective movie productions; and the image of nurses shown in these films, on both sides, presents a conflicting role concept for women in society. Following strategies developed by specialists in film analysis (Bondwell 1995, Uneso 1995, Carmona 1991) a total of 453 movie productions, 360 on the republican side and 93 on the national side, were reviewed. These films were listed in the Spanish National Films Library records. After analyzing the Spanish cinema productions during the Spanish Civil War, data relating to 453 films were identified. The genre included documents, news programs and fiction movies. 77 were produced in 1936, 235 in 1937, 102 in 1938 and 39 in 1939. A tremendous difference exists between the republican productions, 79% of the total, and the national productions. By genres, the types produced on the republican side were: in 1936, 53 documentals, 4 news programs and 9 fiction films; in 1937, 186 documentals, 5 news programs and 19 fiction films; in 1938, 72 documentals, 1 news programs and 2 fiction films; in 1939, 2 documentals and 2 fiction films. On the national side, their productions were: in 1936, 10 documentals and 1 fiction film; in 1937, 22 documentals, 2 news programs and 1 fiction film; in 1938, 19 documentals and 3 news programs; in 1939, 29 documentals and 6 fiction films. During the Spanish Civil War, movies produced by both sides made an effort to reflect their ideal woman as a stereotypical ideal nurse. This ideal nurse showed the values, ideas, aesthetics and prejudices each side held in the war.