Water-pipe smoking promotes epithelial-mesenchymal transition and invasion of human breast cancer cells via ERK1/ERK2 pathways.
With the increasing popularity of water-pipe smoking (WPS), it is critical to comprehend how WPS may affect women's health. The main goal of this study is to identify the potential outcome of WPS on human breast cancer progression. Two breast cancer cell lines, MCF7 and BT20, were used in this investigation. We explored the outcome of WPS on cell morphology and cell invasion using inverted microscope and Biocoat Matrigel invasion chambers. On the other hand, Western blot was employed to study the expression patterns of key control genes of cell adhesion and invasion. Our data reveal that WPS induces epithelial-mesenchymal transition (EMT) of MCF7 and BT20 breast cancer cell lines; thus, WPS enhances cell invasion ability of both cell lines in comparison with their matched controls. More significantly, WPS provokes a down- and up-regulation of E-cadherin and focal adhesion kinase (FAK), respectively, which are important key regulators of cancer progression genes. Finally, our data point out that WPS incites the activation of Erk1/Erk2, which could be behind the stimulation of EMT and invasion as well as the deregulation of E-cadherin and FAK expression. Our data show, for the first time, that WPS initiates EMT and stimulates cell invasion of breast cancer cells, which could incite metastatic development in breast cancer patients. Thus, we believe that further studies, both in vitro and in vivo, are required to elucidate the pathogenic outcome of WPS on cancer progression of several human carcinomas including breast.


Resistance development and insecticide susceptibility in Culex quinquefasciatus against selection pressure of malathion and permethrin and its relationship to cross-resistance towards propoxur.
To determine resistance level and characterize malathion and permethrin resistance in Culex quinquefasciatus, two methods were used namely: WHO procedures of larval bioassay to determine the susceptibility of lethal concentration (LC) and adult bioassay to determine the lethal time (LT) which are resistant to malathion and permethrin. These mosquito strains were bred in the Insectarium, Division of Medical Entomology, IMR. Thousands of late fourth instar larvae which survived the selection pressure to yield 50% mortality of malathion and permethrin were reared and colonies were established from adults that emerged. Larvae from these colonies were then subjected to the subsequent 10 generations in the test undertaken for malathion resistant strain (F61 - F70) and permethrin resistant strain (F54 - F63). Selection pressure at 50% - 70% mortality level was applied to the larvae of each successive generation. The rate of resistance development and resistance ratio (RR) were calculated by LC5 0 for larval bioassay and LT50 value for adult bioassay. The lab bred Cx. quinquefasciatus was used as a susceptible strain for comparison purpose. The adult bioassay test was carried out by using diagnostic dosages of malathion 5.0%, permethrin 0.75% and with propoxur 0.1%. All bioassay results were subjected to probit analysis. The results showed that LC5 0 for both malathion (F61 - F70) and permethrin (F54 - F63) resistant Cx. quinquefasciatus increased steadily to the subsequent 10 generations indicating a marked development of resistance. The adult female malathion resistant strain have developed high resistance level to malathion diagnostic dosage with resistance ratio 9.3 to 9.6 folds of resistance. Permethrin resistance ratio remained as 1.0 folds of resistance at every generation. It was obvious that malathion resistance developing at a higher rate in adult females compared to permethrin. Female adults exposed to 2 hours of exposure period for propoxur 0.1% showed presence of cross-resistance among the both strains of mosquitoes towards propoxur and it was indicated by 70%-100% mortality at 24 hours post-recovery period.


Spatial characteristics and the epidemiology of human infections with avian influenza A(H7N9) virus in five waves from 2013 to 2017 in Zhejiang Province, China.
The five-wave epidemic of H7N9 in China emerged in the second half of 2016. This study aimed to compare the epidemiological characteristics among the five waves, estimating the possible infected cases and inferring the extent of the possible epidemic in the areas that have not reported cases before. The data for the H7N9 cases from Zhejiang Province between 2013 and 2017 was obtained from the China Information Network System of Disease Prevention and Control. The start date of each wave was 16 March 2013, 1 July 2013, 1 July 2014, 1 July 2015 and 1 July 2016. The F test or Pearson's chi-square test were used to compare the characteristics of the five waves. Global and local autocorrelation analysis was carried out to identify spatial autocorrelations. Ordinary kriging interpolation was analyzed to estimate the number of human infections with H7N9 virus and to infer the extent of infections in the areas with no cases reported before. There were 45, 94, 45, 34 and 80 cases identified from the first wave to the fifth, respectively. The death rate was significantly different among the five waves of epidemics (χ2 = 10.784, P = 0.029). The age distribution (F = 0.903, P = 0.462), gender (χ2 = 2.674, P = 0.614) and occupation(χ2 = 19.764, P = 0.407) were similar in each period. Most of the cases were males and farmers. A significant trend (χ2 = 70.328, P<0.001) was identified that showed a growing proportion of rural cases. There were 31 high-high clusters and 3 high-low clusters at the county level among the five waves and 12, 8, 2, 9 and 3 clusters in each wave, respectively. The total cases infected with the H7N9 virus were far more than those that have been reported now, and the affected areas continue to expand. The epidemic in the north of Zhejiang Province persisted in all five waves. Since the second wave, the virus spread to the south areas and central areas. There was an obvious decline in the infected cases in the urban areas, and the epidemics mostly occurred in the rural areas after the fourth wave. The epidemic was relatively dispersed since the third wave had fewer than two cases in most of the areas and showed a reinforcing trend again in the fifth wave. The study revealed that there were few differences in the epidemiologic characteristics among the five waves of the epidemic. However, the areas where the possible epidemic circulated was larger than reported. The epidemic cross-regional expansion continued and mostly occurred in rural areas. Continuous closure of the live poultry market (LPM) is strongly recommended in both rural and urban areas. Illegal and scattered live poultry trading, especially in rural areas, must be forbidden. It is suggested too that a more rigorous management be performed on live poultry trade and wholesale across the area. Health education, surveillance of cases and pathogenicity should also be strengthened.


Apodization and windowing eigenfunctions.
Across a range of spectral estimation problems and beam focusing problems, it is necessary to constrain the properties of a function and its Fourier transform. In many cases, compact functions in both domains are desired, within the theoretical bounds of the uncertainty principle. Recently, a hyperbolic sine function of modified argument and power was found to be an approximate eigenfunction of the Fourier transform operation, and demonstrated useful properties of compactness with low side lobes. The empirical finding of the eigenfunction relationship is explained by comparison with the prolate spheroidal wave functions, which have exact eigenfunction properties, and their usefulness is demonstrated by examples.


Exposure to Low-Dose X-Ray Radiation Alters Bone Progenitor Cells and Bone Microarchitecture.
Exposure to high-dose ionizing radiation during medical treatment exerts well-documented deleterious effects on bone health, reducing bone density and contributing to bone growth retardation in young patients and spontaneous fracture in postmenopausal women. However, the majority of human radiation exposures occur in a much lower dose range than that used in the radiation oncology clinic. Furthermore, very few studies have examined the effects of low-dose ionizing radiation on bone integrity and results have been inconsistent. In this study, mice were irradiated with a total-body dose of 0.17, 0.5 or 1 Gy to quantify the early (day 3 postirradiation) and delayed (day 21 postirradiation) effects of radiation on bone microarchitecture and bone marrow stromal cells (BMSCs). Female BALBc mice (4 months old) were divided into four groups: irradiated (0.17, 0.5 and 1 Gy) and sham-irradiated controls (0 Gy). Micro-computed tomography analysis of distal femur trabecular bone from animals at day 21 after exposure to 1 Gy of X-ray radiation revealed a 21% smaller bone volume (BV/TV), 22% decrease in trabecular numbers (Tb.N) and 9% greater trabecular separation (Tb.Sp) compared to sham-irradiated controls (P < 0.05). We evaluated the differentiation capacity of bone marrow stromal cells harvested at days 3 and 21 postirradiation into osteoblast and adipocyte cells. Osteoblast and adipocyte differentiation was decreased when cells were harvested at day 3 postirradiation but enhanced in cells isolated at day 21 postirradiation, suggesting a compensatory recovery process. Osteoclast differentiation was increased in 1 Gy irradiated BMSCs harvested at day 3 postirradiation, but not in those harvested at day 21 postirradiation, compared to controls. This study provides evidence of an early, radiation-induced decrease in osteoblast activity and numbers, as well as a later recovery effect after exposure to 1 Gy of X-rays, whereas osteoclastogenesis was enhanced. A better understanding of the effects of radiation on osteoprogenitor cell populations could lead to more effective therapeutic interventions that protect bone integrity for individuals exposed to low-dose ionizing radiation.


Influence of age on proprioceptive accuracy in two dimensions.
Slow arm movements were made over a smooth horizontal table at shoulder height. With visual cues excluded, target position was indicated by the index finger of the nonmoving arm touching the underside of the table. 134 subjects (including 26 left-handed subjects) in the age range 8.0 to 24.6 yr. were examined to see if the results were age-related. Accuracy slightly but significantly increased with age. The younger subjects showed a greater accuracy with the dominant hand--a situation which tended to be reversed in the older subjects.


Impact of specific postoperative complications on the outcomes of emergency general surgery patients.
The relative contribution of specific postoperative complications on mortality after emergency operations has not been previously described. Identifying specific contributors to postoperative mortality following acute care surgery will allow for significant improvement in the care of these patients. Patients from the 2005 to 2011 American College of Surgeons' National Surgical Quality Improvement Program database who underwent emergency operation by a general surgeon for one of seven diagnoses (gallbladder disease, gastroduodenal ulcer disease, intestinal ischemia, intestinal obstruction, intestinal perforation, diverticulitis, and abdominal wall hernia) were analyzed. Postoperative complications (pneumonia, myocardial infarction, incisional surgical site infection, organ/space surgical site infection, thromboembolic process, urinary tract infection, stroke, or major bleeding) were chosen based on surgical outcome measures monitored by national quality improvement initiatives and regulatory bodies. Regression techniques were used to determine the independent association between these complications and 30-day mortality, after adjustment for an array of patient- and procedure-related variables. Emergency operations accounted for 14.6% of the approximately 1.2 million general surgery procedures that are included in American College of Surgeons' National Surgical Quality Improvement Program but for 53.5% of the 19,094 postoperative deaths. A total of 43,429 emergency general surgery patients were analyzed. Incisional surgical site infection had the highest incidence (6.7%). The second most common complication was pneumonia (5.7%). Stroke, major bleeding, myocardial infarction, and pneumonia exhibited the strongest associations with postoperative death. Given its disproportionate contribution to surgical mortality, emergency surgery represents an ideal focus for quality improvement. Of the potential postoperative targets for quality improvement, pneumonia, myocardial infarction, stroke, and major bleeding have the strongest associations with subsequent mortality. Since pneumonia is both relatively common after emergency surgery and strongly associated with postoperative death, it should receive priority as a target for surgical quality improvement initiatives. Prognostic and epidemiologic study, level III.


Immediate localized CDKN1A (p21) radiation response after damage produced by heavy-ion tracks.
Using confocal microscopy on immunofluorescence-stained cells, we have investigated the response of CDKN1A (p21), one of the key proteins involved in the DNA damage response pathway, after irradiation with accelerated lead or chromium ions. Each traversal of an accelerated ion leads to the formation of a single, bright focus of the CDKN1A protein in the nuclei of human fibroblasts within 2 min after irradiation at 4 degrees C. This immediate, localized CDKN1A response is specific for particle irradiation with a high linear energy transfer (LET), whereas X irradiation, after a period of induction, yields a diffusely spread pattern, in line with the differences in the microscopic dose deposition pattern of both radiation types. The particle-induced CDKN1A foci persist for several hours until they become diffuse and vanish. These findings suggest that CDKN1A accumulates at the sites of primary DNA damage, possibly mediated by the interaction with proteins involved in DNA repair. Here, for the first time, an immediate biological response confined to the radial extension of low-energy particle tracks ( approximately 1 micrometer) is directly visualized and correlated to ion traversals. This indicates that particle irradiation represents an ideal tool to study the processing of biological damage induced in defined subnuclear regions.


Renoprotective mechanisms of angiotensin II antagonism in experimental chronic renal failure.
We investigated angiotensin II and nitric oxide-cGMP pathway in the development of hypertension and renal damage in chronic experimental nephritis. Rats with autoimmune nephritis were treated for 12 weeks with AT1 receptor antagonist L-158,809 and/or ACE inhibitor captopril given in drinking water. Blood pressure, urinary albumin, and urinary excretion of cGMP were measured. Renal density of ACE, AT1 and AT2 receptors was determined by quantitative in vitro autoradiography. L-158,809, captopril, and their combination decreased blood pressure and normalised urinary albumin excretion rate in rats with nephritis. In L-158,809-treated rats, cGMP excretion was increased compared to the vehicle-treated nephritic group suggesting that the dysfunctional nitric oxide system may be activated by angiotensin antagonism. In nephritic rats, AT1 and AT2 receptor binding densities in renal medulla were decreased, cortical AT receptor expression remained unchanged. Following L-158,809 treatment, both AT1 and AT2 receptor binding was suppressed. Long-term blockade of AT1 receptors in chronic nephritis has beneficial effects both on albuminuria and blood pressure being as effective as ACE inhibition or their combination. The stimulatory effect of AT1 receptor antagonism on cGMP production was not mediated by AT2 receptor-dependent mechanisms suggesting that AT1 receptor blockade per se favours activation of humoral pathways that stimulate cGMP production and potentially contribute to renal protection in chronic nephritis.


Measuring infant movement. Clinical and technological assessment techniques.
Assessment of infant movement patterns is an important evaluation component in the early identification and management of high-risk and handicapped infants. Differences in infant movement patterns may provide early clues to the diagnosis of cerebral palsy and other developmental disabilities. The ultimate goal of early identification of movement disorders is the provision of early intervention services, such as developmental physical therapy. Qualitative analysis of infant movement differences can assist the clinician in developing treatment strategies aimed at facilitating normal movement patterns during the critical period of early central nervous system plasticity. This article presents four measurement strategies for quantifying such qualitative differences. Three clinical assessment tools--the Movement Assessment of Infants, the Bayley Motor Scale, and the Peabody Developmental Motor Scales--are described. Finally, new technological advances for kinematic analysis of infant movement are presented.


Comorbid Conditions Differentiate Rehabilitation Profiles in Traumatic Versus Nontraumatic Brain Injury: A Retrospective Analysis Using a Medical Database.
We examined the relationship between comorbid medical conditions and changes in cognition over the course of rehabilitation following acquired brain injury. In particular, we compared outcomes between traumatic brain injury (TBI) and non-TBI using a retrospective inpatient rehabilitation dataset. We hypothesized that differences by diagnosis would be minimized among subgroups of patients with common comorbid medical conditions. We used the Functional Independence Measure (FIM)-cognition subscale to index changes in cognition over rehabilitation. A decision tree classifier determined the top 10 comorbid conditions that maximally differentiated TBI and non-TBI. Ten subsets of patients were identified by matching on these conditions, in rank order. Data from these subsets were submitted to repeated-measures logistic regression to establish the minimum degree of commonality in comorbid conditions that would produce similar cognitive rehabilitation, regardless of etiology. The TBI group demonstrated a greater increase in ordinal scores over time relative to non-TBI, across all subscales of the FIM-cognition. When both groups were matched on the top 3 symptoms, there were no significant group differences in rehabilitation trajectory in problem-solving and memory domains (Cohen's d range: 0.2-0.4). Comorbid medical conditions explain differences in cognitive rehabilitation trajectories following acquired brain injury beyond etiology.


Clinicians' perceptions of placing a fetal oximetry sensor.
The aim of this study was to determine clinicians' perceptions of placing a fetal oximetry sensor. A survey was developed seeking details of staff category, birth suite experience, previous sensor placement, ease of application and use of epidural analgesia. Following staff feedback, the survey was revised from asking clinicians to rate women's comfort during sensor placement, to a visual analogue scale (VAS) of the clinician's assessment of the women's pain during sensor placement. Survey validity was sought from experts during development and for specific questions using item-total correlation in this analysis. Ease of sensor placement (n = 131) was rated as good or excellent 71% of the time. Clinicians rated women's comfort during sensor placement as good or excellent in 90% of cases. Median pain VAS was 0 (range 0-7.5). Item-total correlations of survey questions were all significant. Fetal oximetry sensor placement was rated well by clinicians in this purpose-designed survey.


[Comparative study of iodine and thyroid gland in biological processes: development and reproduction].
Iodine is an scarce metalloid, essential for life, more abundant in the aquatic ecosystem than in land. Marine species do not need to store and transform the inorganic iodine into organic iodine through the thyroid gland. Adaptation to terrestrial ecosystem has required phylogenetic processes, and animal species have used different strategies to solve the iodine captation problem according to their needs. Jellyfish lack of thyroid gland while lampery developed the thyroid gland during evolution in order to adapt to the terrestrial ecosystem. This review is a comparative phylogenetic study of thyroid gland development and iodine physiology, includes functional disorders, as well as iodine captation ability and requirements during growth and reproduction in different species.


Clinicopathological review of 18 cases of liver allografts lost due to bile duct necrosis.
Clinical, radiographic, and pathological features of 18 patients with biliary necrosis in their explanted liver allografts were reviewed. Twelve patients were men and ages ranged from 27 to 72 years. Indications for initial liver transplant (LT) were viral hepatitis (n = 7), steatohepatitic cirrhosis (n = 3), cryptogenic cirrhosis (n = 3), secondary sclerosing cholangitis (n = 2), primary sclerosing cholangitis (n = 1), biliary atresia (n = 1), and nodular regenerative hyperplasia (n = 1). Donor age ranged from 16 to 75 years. Duct-to-duct biliary anastomoses were fashioned in 13 cases; warm and cold ischemia times were not significantly different from general LT population. Seventeen allograft biopsies after recirculation had no significant findings. Post-LT, clinical and radiographic evaluation indicated biliary strictures (n = 7), bile leak (n = 7), intrahepatic abscess (n = 1), and duodenal perforation (n = 1). Radiographic vascular studies suggested hepatic arterial thrombosis or stenosis in 11 cases. Biopsies prior to retransplantation were performed on 17 patients and showed acute rejection (n = 10), biliary outflow impairment (n = 4), normal histology (n = 2), and centrilobular necrosis (n = 1). Retransplantation was performed 14 to 334 days after initial LT. Pathological examination of explants revealed perihilar duct necrosis in all cases, with bacterial colonies (n = 10) and fungal organisms (n = 2). Arterial thrombi were seen in 10 cases, and two had prominent arteriosclerosis. Infarction and centrilobular necrosis were seen in 9 and 13 cases, respectively. Four explants showed features of biliary outflow impairment. Twelve patients were alive 6 to 18 months following retransplantation. We conclude that post-LT biliary necrosis is associated with ischemia, and such a complication is rarely evident in allograft biopsies. Biliary and vascular imaging studies are essential in evaluating patients for this complication.


Plasmid-based vaccines encoding rat neu and immune stimulatory molecules can elicit rat neu-specific immunity.
DNA vaccines are ideally suited for immunizing against tumor antigens because constructs can be formulated that not only encode the tumor antigen but also encode molecules chosen to improve the ability to elicit an antitumor response. Ligands expressed on antigen-presenting cells associated with stimulating a robust T-cell response are excellent candidates for inclusion in a DNA vaccine. Mice transgenic for the HER-2/neu homologue, rat neu, were immunized with full-length rat neu cDNA given alone or in combination with plasmids encoding costimulatory molecules CD80 or CD86 and the ligand for CD137 (CD137L). Intradermal injection of the plasmid constructs resulted in both plasmid transcript and antigen protein expression being detected in lymph nodes draining the injection site. Immunization with plasmids encoding the neu antigen along with plasmids encoding CD137L and either CD80 or CD86 resulted in the generation of neu-specific antibodies that induced phopshorylation of the neu tyrosine kinase and inhibited the growth of cultured tumor cells overexpressing neu. Survival of animals was significantly prolonged after immunization with vaccines encoding neu together with the costimulatory molecules. Although tumors eventually occurred in the vaccinated animals, they were markedly infiltrated with CD4+ T cells. DNA vaccines encoding neu, when given in combination with both CD137L and either CD80 or CD86, can induce cellular and humoral immunity and result in an antitumor effect.


51V-n.m.r. analysis of the binding of vanadium(V) oligoanions to sarcoplasmic reticulum.
The binding of mono- and oligo-vanadates to sarcoplasmic reticulum was analysed by 51V-n.m.r. spectroscopy. The observations indicate that, in addition to monovanadate, the di-, tetra- and deca-vanadates are also bound to sarcoplasmic-reticulum membranes with high affinity. The binding of the vanadate oligoanions may explain some of the effects of vanadates on the conformation and crystallization of Ca2+-transport ATPase.


Secreted frizzled related proteins: Implications in cancers.
The Wnt (wingless-type) signaling pathway plays an important role in embryonic development, tissue homeostasis, and tumor progression becaluse of its effect on cell proliferation, migration, and differentiation. Secreted frizzled-related proteins (SFRPs) are extracellular inhibitors of Wnt signaling that act by binding directly to Wnt ligands or to Frizzled receptors. In recent years, aberrant expression of SFRPs has been reported to be associated with numerous cancers. As gene expression of SFRP members is often lost through promoter hypermethylation, inhibition of methylation through the use of epigenetic modifying agents could renew the expression of SFRP members and further antagonize deleterious Wnt signaling. Several reports have described epigenetic silencing of these Wnt signaling antagonists in various human cancers, suggesting their possible role as tumor suppressors. SFRP family members thus come across as potential tools in combating Wnt-driven tumorigenesis. However, little is known about SFRP family members and their role in different cancers. This review comprehensively covers all the available information on the role of SFRP molecules in various human cancers.


In vivo immune selection of a Moloney murine leukemia virus-induced tumor results in the loss of viral- but not tumor-associated antigens.
A protocol of in vivo immune selection has been used to isolate a variant of the Moloney murine leukemia virus (MuLV)-induced tumor MBL-2. Characterization of the tumor variant indicated that selection resulted in the isolation of a cell which is incapable of producing infectious virus and no longer capable of synthesizing viral proteins. Although the failure to express viral Ag has rendered the variant tumor cells resistant to lysis by CTL specific for MuLV viral Ag, the variant tumor cells retained their susceptibility to lysis by CTL which appear to be directed against an MuLV-induced tumor-associated Ag. The data indicate that the expression of nonviral tumor-associated Ag by MBL-2 is not dependent upon continued viral gene expression.


Larvicidal Potential of Five Selected Dragonfly Nymphs in Sri Lanka over Aedes aegypti (Linnaeus) Larvae under Laboratory Settings.
Limitations in breeding source reduction practices, development of insecticide resistance in mosquitoes, and ill effects of chemical controlling methods on human and ecosystem health have motivated Sri Lankan authorities working for dengue control to seek for alternative, ecofriendly, and sustainable approaches for controlling of Aedes vectors, to manage dengue epidemics. The present study attempted to investigate the predation efficiency of locally available dragonfly nymphs over Aedes aegypti under laboratory conditions, aiming to evaluate the potential of using dragonflies as biocontrol agents against dengue. Nymphal stages of five locally abundant dragonfly species were collected from different stagnated water bodies in Belihuloya area. After morphological identification, a well grown individual of each species was starved for 12 hours and introduced into a glass tank containing 1L of pond water with 200 larvae (4th instar) of Aedes aegypti. Number of larvae survived in the tank was enumerated hourly up to 48 hours. In case where >75% of larvae are consumed by dragonfly nymphs, additional Ae. aegypti larvae were introduced into such tanks. Experiment was repeated for five times. Same procedure was followed with different stages of growth of the dragonfly nymphs characterized by the highest predation rate. General Linear Model followed by Tukey's pairwise comparison was used for statistical analysis. The predation rates of different dragonfly species varied significantly (p<0.05), whereby Anax indicus (110±7.14 per day) indicated the highest, followed by Pantala flavescens (54.07±5.15) and Gynacantha dravida (49.00±11.89), while Tholymis tillarga (23.47±2.48) had the lowest. Further, significant variations in the larval predation were found among different maturity stages (10-20; 25-35; and 35-45 mm in body length) of Ana. indicus (p<0.05). Regardless of statistical significance, a relatively higher larvicidal activity was observed at dusk than in dawn. Conclusion. Ana. indicus, which is characterized by the highest predation rate, and P. flavescens that has the widest geographical distribution within Sri Lanka along with a notable predation efficacy could be recommended as potential candidates for field trials in biological control of dengue outbreaks via suppression of Ae. aegypti larvae.


Severe intellectual disability and autistic features associated with microduplication 2q23.1.
We report on two patients with developmental delay, hypotonia, and autistic features associated with duplications of chromosome region 2q23.1-2q23.2 detected by chromosome microarray analysis. The duplications include one OMIM Morbid Map gene, MBD5, as well as seven known RefSeq genes (ACVR2A, ORC4L, EPC2, KIF5C, MIR1978, LYPD6B, and LYPD6). MBD5 lies in the minimum area of overlap of the 2q23.1 microdeletion syndrome. This report provides the first detailed clinical examination of two individuals with a duplication of this region and suggests that brain development and cognitive function may be affected by an increased dosage of the genes involved.


Traditional Small-Size Citrus from Taiwan: Essential Oils, Bioactive Compounds and Antioxidant Capacity.
Background: The calamondin (Citrus microcarpa Bunge) and the kumquat (Fortunella crassifolia Swingle) are two small-size citrus fruits that have traditionally been consumed in Taiwan; however, there has been a lack of scientific research regarding the active compounds and functionalities of these fruits. Methods: Analysis of volatile composition of essential oil and phytosterol was carried out using Gas Chromatography-Mass Spectrometry (GC-MS). Flavonoid and limonoid were analyzed by High Performance Liquid Chromatography (HPLC). Moreover, antioxidant capacity from their essential oils and extracts were assessed in vitro. Results: The compositions of the essential oils of both fruits were identified, with the results showing that the calamondin and kumquat contain identified 43 and 44 volatile compounds, respectively. In addition, oxygenated compounds of volatiles accounted for 4.25% and 2.04%, respectively, consistent with the fact that oxygenated compounds are generally found in high content in citrus fruits. In terms of flavonoids, the calamondin exhibited higher content than the kumquat, with disomin-based flavonoids being predominant; on the other hand, phytosterol content of kumquat was higher than that of calamondin, with amyrin being the dominant phytosterol. Both of them contain high amounts of limonoids. The ethanol extracts and essential oils of small-sized citrus fruits have been shown to have antioxidant effects, with those effects being closely related to the flavonoid content of the fruit in question. Conclusions: The present study also reviewed antioxidant activity in terms of specific bioactive compounds in order to find the underlying biological activity of both fruits. The calamondin and kumquat have antioxidant effects, which are in turn very important for the prevention of chronic diseases.


Role for DYRK family kinases on regulation of apoptosis.
The cellular response to a variety of stress including DNA damage is involved in cell cycle arrest, activation of DNA repair, and in the event of irreparable damage, induction of apoptosis. However, the signals that determine cell fate, that is, survival or apoptosis, are largely unknown. Accumulating studies have revealed that dual-specificity tyrosine-regulated kinases (DYRKs) play key roles on cell proliferation and apoptosis induction. In particular, DYRK2 translocates from the cytoplasm into the nucleus following genotoxic stress. DYRK2 is then activated by ATM and induce apoptosis by phosphorylating p53 at Ser46. Importantly, whereas precise regulation of these kinases remain uncertain, this mechanism has consequences for cell proliferation, differentiation, or apoptosis. This progress review highlights recent efforts demonstrating that DYRKs could be novel and essential regulatory molecules for the regulation of cell fate including apoptosis.


Transglutaminases, thioredoxins and protein disulphide isomerase: diverse enzymes with a common goal of cross-linking proteins in lower organisms.
Prokaryotes and various eukaryotes have remarkable ability to survive under adverse physiologic conditions and protect themselves from environmental stresses. An important mechanism by which they accomplish this is by synthesizing rigid and biochemically inert structures around them. In general, these structures are highly stable and resistant to mechanical and chemical insults. Biochemically, they are composed of complex carbohydrates, such as chitin and heavily crosslinked scaffold of proteins to form complex structures, such as sheath, cuticle, and epicuticle. Transglutaminases (TGases) are a family of enzymes that share catalytic function with thioredoxin and protein disulphide isomerases (PDI) and catalyze protein crosslink reaction by establishing epsilon-(gamma-glutamyl)lysine isopeptide bonds. The isopeptide bonds thus formed are of great physiologic significance because once formed, they cannot be hydorlysed by any known enzymes of the eukaryote system and exhibit high resistance to reducing agents, detergents, and chaotropic agents. Therefore, it is likely that protective structures viz., sheath, cuticle, epicuticle, and viral core proteins synthesized by microorganisms involve active participation of TGases. In this review, we briefly describe the current knowledge of non-mammalian TGases and their possible role in growth, development, and survival of small organisms. Special reference is made to filarial nematode and bacterial TGases since they are the most well-characterized and studied enzymes among non-mammalian TGases.


Metabolome profile variation in Azolla filiculoides exposed to Bisphenol A assists in the identification of stress-responsive metabolites.
This study attempted to explore the metabolome profile of Azolla filiculoides subjected to two different concentrations of BPA (1 and 30 mg L-1) in congruence with two different durations (3 and 9 days) of treatment. Bisphenol A (BPA) is a ubiquitously occurring environmental pollutant that imparts acute toxicity in aquatic plants. Therefore, studying the variations in the fern metabolome profile and identifying stress-responsive metabolites can help develop criteria for assessing the aquatic ecosystem. In recent times, metabolomics has drawn attention for its ability to detect biochemical processes and help link plant responses with environmental stresses. However, the studies concerning the metabolome profile of A. filiculoides exposed to environmental contaminants are limited. In the present study, the untargeted metabolomics study allowed the detection of a large array of metabolites, with 767 shared metabolites representing 41 crucial pathways. Exposure to 30 mg L-1 BPA seemingly disrupted the primary metabolism of the fern and induced a shift toward defense-related pathways. Additionally, BPA stress triggered the expression of metabolites like 3,4-dihydroxyphenylglycol, perillic acid, and perillaldehyde in BPA_L3 (1 mg L-1 for 3 days) and BPA_L9 (1 mg L-1 for 9 days) samples indicating protective mechanism of the plants. Conversely, the BPA_H3 (30 mg L-1 for 3 days) and BPA_H9 (30 mg L-1 for 9 days) samples expressed a distinct set of markers like luteolin, 3-hydroxyanthranilic acid, cinnamaldehyde, and l-DOPA indicating the onset of senescence and apoptosis related pathways can help in the health assessment of freshwater ecosystems and also appraisal of ecotoxicological risks imposed by BPA.


Food for thought: Leptin regulation of hippocampal function and its role in Alzheimer's disease.
Accumulating evidence indicates that diet and body weight are important factors associated with Alzheimer's disease (AD), with a significant increase in AD risk linked to mid-life obesity, and weight loss frequently occurring in the early stages of AD. This has fuelled interest in the hormone leptin, as it is an important hypothalamic regulator of food intake and body weight, but leptin also markedly influences the functioning of the hippocampus; a key brain region that degenerates in AD. Increasing evidence indicates that leptin has cognitive enhancing properties as it facilitates the cellular events that underlie hippocampal-dependent learning and memory. However, significant reductions in leptin's capacity to regulate hippocampal synaptic function occurs with age and dysfunctions in the leptin system are associated with an increased risk of AD. Moreover, leptin is a potential novel target in AD as leptin treatment has beneficial effects in various models of AD. Here we summarise recent advances in leptin neurobiology with particular focus on regulation of hippocampal synaptic function by leptin and the implications of this for neurodegenerative disorders like AD. This article is part of the Special Issue entitled 'Metabolic Impairment as Risk Factors for Neurodegenerative Disorders.'


JNK-mediated phosphorylation of paxillin in adhesion assembly and tension-induced cell death by the adenovirus death factor E4orf4.
The adenovirus type 2 Early Region 4 ORF4 (E4orf4) protein induces a caspase-independent death program in tumor cells involving changes in actin dynamics that are functionally linked to cell killing. Because an increase in myosin II-based contractility is needed for the death of E4orf4-expressing cells, we have proposed that alteration of cytoskeletal tension is part of the signals engaging the death pathway. Yet the mechanisms involved are poorly defined. Herein, we show that the Jun N-terminal kinase JNK is activated in part through a pathway involving Src, Rho, and ROCK (Rho kinase) and contributes to dysregulate adhesion dynamics and to kill cells in response to E4orf4. JNK supports the formation of atypically robust focal adhesions, which are bound to the assembly of the peculiar actomyosin network typifying E4orf4-induced cell death and which are required for driving nuclear condensation. Remarkably, the dramatic enlargement of focal adhesions, actin remodeling, and cell death all rely on paxillin phosphorylation at Ser-178, which is induced by E4orf4 in a JNK-dependent way. Furthermore, we found that Ser-178-paxillin phosphorylation is necessary to decrease adhesion turnover and to enhance the time residency of paxillin at focal adhesions, promoting its recruitment from an internal pool. Our results indicate that perturbation of tensional homeostasis by E4orf4 involves JNK-regulated changes in paxillin adhesion dynamics that are required to engage the death pathway. Moreover, our findings support a role for JNK-mediated paxillin phosphorylation in adhesion growth and stabilization during tension signaling.


[Combined cycloferon treatment of tuberculosis in patients infected with HIV].
Outcome of treatment of 102 patients co-infected with HIV and tuberculosis hospitalized at Second Municipal Tuberculosis Hospital (St. Petersburg) is analyzed. The median period of follow up was 8 weeks. In order to select the optimum treatment scheme, all patients were divided into three groups (I-III). Group I (51 patients) received, in addition to antituberculosis medications, cycloferon 600 mg tablets on alternate days. Group II (16 patients) received alpha and gamma interferons (Interal injection, 3 MU and Ingaron injection, 500,000 on alternate days) in combination with antituberculosis medications. Control group III (35 patients) received antituberculosis treatment plus placebo. Assessment of the outcome of TB treatment upon 8 weeks of follow up showed a remarkable improvement of clinical and immunological conditions in the patients who received interferons and cycloferon.


Detection of precipitating antibodies to Histoplasma capsulatum by counterimmunoelectrophoresis.
The technique of counterimmunoelectrophoresis was evaluated for its usefulness in the detection of precipitating antibodies to Histoplasma capsulatum, designated h and m precipitin bands. Forty-four patients with active histoplasmosis had either m bands or both h and m bands. The h precipitin band occurred primarily in patients with disseminated disease, chronic pulmonary disease, or mediastinal lymphadenopathy of several months' duration; with resolution of the infection, this antibody disappeared. The m band appeared earlier in the course of histoplasmosis and persisted for months to years after resolution of the infection. Antibodies detected by counterimmunoeelectrophoresis were in the immunoglobin G class and were more specific for histoplasmosis than those detected by the complement fixation test. Only one false positive h band occurred in a total of 81 sera tested (specimens from 24 healthy control subjects and 57 patients with other diseases). Counterimmunoelectrophoresis was both more sensitive and quicker than immunodiffusion at detecting h and m antibodies. Combining this assay with standard complement fixation tests and fungal culture methods should lead to the accurate, rapid diagnosis of histoplasmosis.


FOCUS 2: a randomized, double-blinded, multicentre, Phase III trial of the efficacy and safety of ceftaroline fosamil versus ceftriaxone in community-acquired pneumonia.
Ceftaroline (active form of the prodrug ceftaroline fosamil) is a novel cephalosporin with activity against pathogens commonly associated with community-acquired pneumonia (CAP), including Streptococcus pneumoniae and Gram-negative pathogens. This randomized, double-blind, Phase III study evaluated the efficacy and safety of ceftaroline fosamil in treating patients with CAP. The primary objective was to determine non-inferiority [lower limit of 95% confidence interval (CI) ≥ -10%] of clinical cure rates achieved with ceftaroline fosamil compared with those achieved with ceftriaxone in the clinically evaluable (CE) and modified intent-to-treat efficacy (MITTE) populations. Patients hospitalized in a non-intensive care unit setting with CAP of Pneumonia Outcomes Research Team (PORT) risk class III or IV requiring intravenous (iv) therapy were randomized (1:1) to receive 600 mg of ceftaroline fosamil iv every 12 h or 1 g of ceftriaxone iv every 24 h. Clinical cure, microbiological response, adverse events (AEs) and laboratory tests were assessed. FOCUS 2 registration number NCT00509106 (http://clinicaltrials.gov/ct2/show/NCT00509106). The study enrolled 627 patients, 315 of whom received ceftaroline fosamil and 307 of whom received ceftriaxone. Patients in both treatment groups had comparable baseline characteristics. Clinical cure rates were as follows: CE population, 82.1% (193/235) for ceftaroline fosamil and 77.2% (166/215) for ceftriaxone [difference (95% CI), 4.9% (-2.5, 12.5)]; and MITTE population, 81.3% (235/289) for ceftaroline fosamil and 75.5% (206/273) for ceftriaxone [difference (95% CI), 5.9% (-1.0, 12.7)]. Clinical cure rates for CAP caused by S. pneumoniae in the microbiological MITTE (mMITTE) population were 83.3% (35/42) and 70.0% (28/40) for ceftaroline fosamil and ceftriaxone, respectively. Ceftaroline fosamil and ceftriaxone were well tolerated, with similar rates of AEs, serious AEs, deaths and discontinuations due to an AE. The most common AEs for ceftaroline fosamil-treated patients were diarrhoea, headache, hypokalaemia, insomnia and phlebitis, and the most common AEs for ceftriaxone-treated patients were diarrhoea, insomnia, phlebitis and hypertension. Ceftaroline fosamil achieved high clinical cure and microbiological response rates in patients hospitalized with CAP of PORT risk class III or IV. Ceftaroline fosamil was well tolerated, with a safety profile that is similar to that of ceftriaxone and other cephalosporins. Ceftaroline fosamil is a promising agent for the treatment of CAP.


Neuro-genetic optimization of temperature control for a continuous flow polymerase chain reaction microdevice.
In this study, a hybridized neuro-genetic optimization methodology realized by embedding finite element analysis (FEA) trained artificial neural networks (ANN) into genetic algorithms (GA), is used to optimize temperature control in a ceramic based continuous flow polymerase chain reaction (CPCR) device. The CPCR device requires three thermally isolated reaction zones of 94 degrees C, 65 degrees C, and 72 degrees C for the denaturing, annealing, and extension processes, respectively, to complete a cycle of polymerase chain reaction. The most important aspect of temperature control in the CPCR is to maintain temperature distribution at each reaction zone with a precision of +/-1 degree C or better, irrespective of changing ambient conditions. Results obtained from the FEA simulation shows good comparison with published experimental work for the temperature control in each reaction zone of the microfluidic channels. The simulation data are then used to train the ANN to predict the temperature distribution of the microfluidic channel for various heater input power and fluid flow rate. Once trained, the ANN analysis is able to predict the temperature distribution in the microchannel in less than 20 min, whereas the FEA simulation takes approximately 7 h to do so. The final optimization of temperature control in the CPCR device is achieved by embedding the trained ANN results as a fitness function into GA. Finally, the GA optimized results are used to build a new FEA model for numerical simulation analysis. The simulation results for the neuro-genetic optimized CPCR model and the initial CPCR model are then compared. The neuro-genetic optimized model shows a significant improvement from the initial model, establishing the optimization method's superiority.


Structure and interactions of the first three RNA recognition motifs of splicing factor prp24.
The essential Saccharomyces cerevisiae pre-messenger RNA splicing protein 24 (Prp24) has four RNA recognition motifs (RRMs) and facilitates U6 RNA base-pairing with U4 RNA during spliceosome assembly. Prp24 is a component of the free U6 small nuclear ribonucleoprotein particle (snRNP) but not the U4/U6 bi-snRNP, and so is thought to be displaced from U6 by U4/U6 base-pairing. The interaction partners of each of the four RRMs of Prp24 and how these interactions direct U4/U6 pairing are not known. Here we report the crystal structure of the first three RRMs and the solution structure of the first two RRMs of Prp24. Strikingly, RRM 2 forms extensive inter-domain contacts with RRMs 1 and 3. These contacts occupy much of the canonical RNA-binding faces (beta-sheets) of RRMs 1 and 2, but leave the beta-sheet of RRM 3 exposed. Previously identified substitutions in Prp24 that suppress mutations in U4 and U6 spliceosomal RNAs cluster primarily in the beta-sheet of RRM 3, but also in a conserved loop of RRM 2. RNA binding assays and chemical shift mapping indicate that a large basic patch evident on the surface of RRMs 1 and 2 is part of a high affinity U6 RNA binding site. Our results suggest that Prp24 binds free U6 RNA primarily with RRMs 1 and 2, which may remodel the U6 secondary structure. The beta-sheet of RRM 3 then influences U4/U6 pairing through interaction with an unidentified ligand.


Buprenorphine Microdosing Cross Tapers: A Time for Change.
Buprenorphine is a partial opioid agonist that is Food and Drug Administration (FDA) approved to treat chronic pain and opioid use disorder (OUD). The national prescribing guidelines in the United States (US) recommend that patients transitioning from full opioid agonists to buprenorphine first undergo 12 or more hours of active opioid withdrawal, in order to avoid buprenorphine-precipitated opioid withdrawal. This opioid-free period imposes a significant barrier for many patients. Evidence is accumulating that using microdoses of buprenorphine to cross taper from full-agonist opioids to buprenorphine is a safe and effective way to avoid opioid withdrawal and uncontrolled pain. This microdose cross-tapering strategy is already being used across the US. The US prescribing guidelines and buprenorphine training would benefit from acknowledging this new approach. Additionally, to facilitate this strategy, the FDA should approve transdermal buprenorphine formulations for OUD and manufacturers could produce lower dose formulations of sublingual buprenorphine. The time has come for us to embrace buprenorphine microdosing cross tapers as a new standard of care.


Neuroticism polygenic risk score predicts 20-year burden of depressive symptoms for Whites but not Blacks.
Black-White differences are reported in social, psychological, behavioral, medical, and biological correlates of depression. This study was conducted to compare Black and White older adults for the association between neuroticism polygenic risk score (N-PRS) and chronicity of depressive symptoms over 20 years. Data came from the Health and Retirement Study (HRS), 1990 - 2012, a nationally representative sample of Americans above age 50. Current analysis followed 9,249 individuals (7,924 Whites and 1,325 Blacks) for up to 22 years. Depressive symptoms were measured every two years between 1992 and 2012 using the 8-item Center for Epidemiological Studies-Depression Scale (CES-D-8). The independent variable was N-PRS. The dependent variable was average depressive symptoms between 1992 and 2012. Linear regression was used for data analysis. In the pooled sample, higher N-PRS was associated with higher average depressive symptoms over the 20-year follow up period [b=0.01, 95%CI=0.00 to 0.04], net of all covariates. We also found an interaction between race and N-PRS [b=-0.02, 95%CI=-0.03 to 0.00], suggesting a stronger effect of N-PRS on 20-year average depressive symptoms for Whites than Blacks. Based on our race-specific linear regression models, higher N-PRS was associated with higher depressive symptoms from 1992 to 2012 for Whites [b=0.01, 95%CI=0.01 to 0.02] but not Blacks [b=0.00, 95%CI=-0.02 to 0.02]. Black and White older adults may differ in the salience of the existing N-PRS for depressive symptoms, which better reflects the burden of depression for Whites than Blacks. This may be because the existing PRSs are derived from mostly or exclusively White samples, limiting their applicability in other race groups. Racial variation in psychosocial, clinical, and biological correlates of depression needs further research.


[Ultrasonically-guided percutaneous puncture and drainage of a splenic abscess caused by Salmonella heidelberg in a child].
In a boy aged 9 years and 11 months pain developed in the left upper abdomen following Salmonella enteritis infection. Purulent splenitis was diagnosed by sonography. Ultrasonically guided percutaneous puncture and drainage, and appropriate chemotherapy led to a marked improvement in the clinical picture and finally to complication-free remission. In a review of the literature the problems associated with purulent splenitis and the possibility of treating it by percutaneous puncture are pointed out. Purulent splenitis should be considered in all patients with temperatures and unexplained abdominal pain.


Treatment for T1a Renal Cancer Substratified by Size: "Less is More".
Due to the widespread use of computerized tomography, the diagnosis of small renal cancers (3 cm or less) within the T1a classification continues to increase. Current treatment of these tumors includes radical nephrectomy, partial nephrectomy and thermal ablation. We used the SEER (Surveillance, Epidemiology, and End Results) Program to compare treatment modalities for these cancers based on 1 cm increments in tumor size. We examined overall survival, cancer specific survival, survival from cardiovascular disease and race based treatment disparities. In the SEER database we identified 17,716 renal cancers 3 cm or less diagnosed from 2005 to 2010 treated with radical nephrectomy, partial nephrectomy or thermal ablation. Overall survival, cancer specific survival and cardiovascular survival were determined for each treatment group, and then substratified by size in centimeters, tumor grade, age, geographical location and ethnicity. Survival was analyzed using Kaplan-Meier methods, multivariate proportional hazards models and a propensity score weighted approach. Overall survival, cancer specific survival and cardiovascular survival were better for partial nephrectomy than radical nephrectomy in all circumstances. Thermal ablation showed equivalent overall survival to partial nephrectomy for tumors 2 cm or less. Notably, radical nephrectomy for renal tumors 3 cm or less was applied in a disparately larger number of black patients (OR 1.63, 95% CI 1.47-1.81) and Hispanic patients (OR 1.28, 95% CI 1.14-1.44). Radical nephrectomy should be avoided for all tumors 3 cm or less. For renal cancers 2 cm or less partial nephrectomy and thermal ablation are equally effective. For tumors 2.1 to 3 cm partial nephrectomy is better than thermal ablation. We identified significant racial treatment disparities that negatively impact survival in black and Hispanic patients.


The coronary collateral circulation in normal goats.
This study was undertaken to evaluate the coronary collateral circulation of the goat. Year old, castrated male goats were anesthetized with sodium pentobarbital. A branch of the left circumflex coronary artery was dissected and a snare placed around it. Regional myocardial blood flow was measured by injecting colored microspheres into the left atrium before and during 3 hr of occlusion of this coronary artery. The Area At Risk for infarction, as defined by a left atrial injection of Brilliant Green dye, was divided into a central Ischemic zone and a peripheral Ischemic Border zone. The degree of overlap in the blood flow distributions between the risk and the nonrisk areas was quantitatively assessed by injecting microspheres directly into the artery to the Area At Risk. Baseline blood flow to the normally perfused goat myocardium was 1.13 +/- 0.18 ml/min/g (mean +/- SE). Following occlusion, the flow to the Ischemic zone was 0.07 +/- 0.03 ml/min/g and to the Ischemic Border zone was 0.31 +/- 0.18 ml/min/g. Flow to either zone did not increase during the 3-hr observation period. Overlap at the perimeter of the risk and nonrisk areas was approximately 22%. We conclude that flow to the Ischemic zone is low because the goat has few native collateral blood vessels, and that flow to the Ischemic Border zone is significantly affected by overlap with normal myocardium.


GalphaL1 (Galpha14) couples the opioid receptor-like1 receptor to stimulation of phospholipase C.
In most tissues and cells the opioid receptor-like (ORL1) receptor regulates effectors primarily through the pertussis toxin (PTX)-sensitive guanine nucleotide-binding regulatory proteins (G proteins) Gi/Go. Many Gi-coupled receptors possess additional capability to interact with one or more PTX-insensitive G proteins. Using the betagamma-induced stimulation of type 2 adenylyl cyclase as a readout, we screened the ability of ORL1 receptor to interact with a panel of PTX-insensitive G proteins. In the presence of PTX, activation of the ORL1 receptor resulted in the stimulation of type 2 adenylyl cyclase only in HEK 293 cells coexpressing the alpha subunit of Gz, G12, G14, or G16, but not in cells coexpressing G11, G13, or Gq. Coupling to both Gz and G16 was expected because close relatives of the ORL1 receptor, the opioid receptors, are known to couple productively to these G proteins. ORL1 receptor coupling to either G12 or G14 has not been demonstrated. As predicted by the type 2 adenylyl cyclase assays, activation of the ORL1 receptor resulted in the formation of inositol phosphates in COS-7 cells transiently cotransfected with Galpha14. The ORL1 receptor-mediated stimulation of phospholipase C was found to be Galpha14 dependent, agonist dose dependent, ligand selective, and PTX insensitive. We conclude that G14 can link the ORL1 receptor to regulation of phopholipase C.


Epicardial adipose tissue reflects the presence of coronary artery disease: comparison with abdominal visceral adipose tissue.
Accumulation of visceral adipose tissue is associated with a risk of coronary artery disease (CAD). The aim of this study was to examine whether different types of adipose tissue depot may play differential roles in the progression of CAD. Consecutive 174 patients who underwent both computed tomography (CT) and echocardiography were analyzed. Cardiac and abdominal CT scans were performed to measure epicardial and abdominal visceral adipose tissue (EAT and abdominal VAT, resp.). Out of 174 patients, 109 and 113 patients, respectively, presented coronary calcification (CC) and coronary atheromatous plaque (CP). The EAT and abdominal VAT areas were larger in patients with CP compared to those without it. Interestingly, the EAT area was larger in patients with CC compared to those without CC, whereas no difference was observed in the abdominal VAT area between patients with CC and those without. Multivariable logistic regression analysis revealed that the presence of echocardiographic EAT was an independent predictor of CP and CC, but the abdominal VAT area was not. These results suggest that EAT and abdominal VAT may play differential pathological roles in CAD. Given the importance of CC and CP, we should consider the precise assessment of CAD when echocardiographic EAT is detected.


An insulin-independent mechanism of intrathecal morphine-induced hypoglycemia in mice: mediation through a central alpha-2 adrenergic pathway.
Studies on the mechanism of the hypoglycemia caused by the i.t. administration of morphine (40 micrograms) to nonfasted, unanesthetized mice included assessment of effects of i.t. morphine on circulating levels of immunoreactive insulin and glucagon, on body temperature, as well as testing the effects of i.p. pretreatment with antagonists of cholinergic, beta adrenergic, alpha-1 adrenergic, alpha-2 adrenergic, serotonergic and opioid receptors. Serum levels of insulin were not significantly affected at 30 and 60 min after i.t. morphine, but plasma glucagon levels were significantly increased at both time points. Rectal temperature decreased significantly in mice given either saline or morphine i.t., but there was no significant difference between the two groups over time. Atropine (2 mg/kg), propranolol (10 mg/kg), prazosin (2 mg/kg) and methysergide (2 mg/kg) did not affect morphine-induced hypoglycemia. Naloxone (20 mg/kg) antagonized i.t. morphine. Mecamylamine (2 mg/kg) produced a moderate hypoglycemia in control mice, which appeared to be additive to that caused by i.t. morphine. Yohimbine (1-4 mg/kg i.p.) also produced a moderate hypoglycemia in control mice, but caused a dose-related antagonism of i.t. morphine. The centrally active alpha-2 antagonist, L-657,743 (2S-trans)-1,3,4,5',6,6',7,12b-octahydro-1',3'-dimethylspiro -(2H-benzofuro- (2,3-a)quinolizine-2,4'(1'H)pyrimidin)-2-(3'H)-one hydrochloride) (4-20 mg/kg i.p.), but not the peripherally selective alpha-2 antagonist, L-659,066 (2R-trans)-N-(2-(1,3,4,6,7,12b-he xahydro-2'- oxospiro(2H-benzofuro(2,3-a) quinolizine-2,4-imidazolidin)-3'-yl)ethyl)methanesulfonamide hydrochloride) (4-20 mg/kg i.p.), caused a dose-related antagonism of i.t. morphine. The i.t. coadministration of yohimbine (2 or 10 micrograms) or L-659,066 (10 micrograms) with morphine exhibited no antagonism.(ABSTRACT TRUNCATED AT 250 WORDS)


The child's first dental visit. Age, reasons, oral health status and dental treatment needs among children in Southern Poland.
The aim of this study was to determine the age at and the reasons for the child's first dental visit, and to assess the oral health status and treatment needs in the analysed group of paediatric patients. The study involved 320 children (154 girls and 166 boys) aged between 0.7 months and 13.5 years, visiting the dentist for the first time. All parents/legal guardians of the study participants gave written informed consent for participation in the study. Data on the child's age and reason for the dental visit were collected from interviews with parents. The state of oral health and dental treatment needs were assessed based on clinical examination, according to recommendations of the World Health Organization. collected data were entered into an Excel spreadsheet and analysed using IBM SPSS software (version 24). Research hypotheses were verified using the Chi-square independence test at the level of statistical significance p<0.05. The mean age of children at their first dental visit was 3.79 years (+/- 1.82 years). The most common reasons (60%) for the first dental visit were pain followed by dental caries (33.1%) and the presence of decayed teeth (26.9%). The frequency of caries in the population was 75.9%, which means that only one out of four examined children was free from dental caries. Only 23.1% of patients did not require dental treatment and as many as 76.9% of the studied population needed dental treatment. Polish children make their first dental visit too late (usually at the age of 4 years) in relation to medical recommendations (between 6 and 12 months of life). The predominant reason for the child's first dental visit is caries and its complications. The results of this study indicate the bad oral health of Polish children making their first dental visit and low health awareness of parents and guardians.


The test-retest reliability of anatomical co-ordinate axes definition for the quantification of lower extremity kinematics during running.
Three-dimensional (3-D) kinematic analyses are used widely in both sport and clinical examinations. However, this procedure depends on reliable palpation of anatomical landmarks and mal-positioning of markers between sessions may result in improperly defined segment co-ordinate system axes which will produce in-consistent joint rotations. This had led some to question the efficacy of this technique. The aim of the current investigation was to assess the reliability of the anatomical frame definition when quantifying 3-D kinematics of the lower extremities during running. Ten participants completed five successful running trials at 4.0 m·s(-1) ± 5%. 3-D angular joint kinematics parameters from the hip, knee and ankle were collected using an eight camera motion analysis system. Two static calibration trials were captured. The first (test) was conducted prior to the running trials following which anatomical landmarks were removed. The second was obtained following completion of the running trials where anatomical landmarks were re-positioned (retest). Paired samples t-tests were used to compare 3-D kinematic parameters quantified using the two static trials, and intraclass correlations were employed to examine the similarities between the sagittal, coronal and transverse plane waveforms. The results indicate that no significant (p>0.05) differences were found between test and retest 3-D kinematic parameters and strong (R(2)≥0.87) correlations were observed between test and retest waveforms. Based on the results obtained from this investigation, it appears that the anatomical co-ordinate axes of the lower extremities can be defined reliably thus confirming the efficacy of studies using this technique.


Longitudinal effects on mental health of moving to greener and less green urban areas.
Despite growing evidence of public health benefits from urban green space there has been little longitudinal analysis. This study used panel data to explore three different hypotheses about how moving to greener or less green areas may affect mental health over time. The samples were participants in the British Household Panel Survey with mental health data (General Health Questionnaire scores) for five consecutive years, and who relocated to a different residential area between the second and third years (n = 1064; observations = 5320). Fixed-effects analyses controlled for time-invariant individual level heterogeneity and other area and individual level effects. Compared to premove mental health scores, individuals who moved to greener areas (n = 594) had significantly better mental health in all three postmove years (P = .015; P = .016; P = .008), supporting a "shifting baseline" hypothesis. Individuals who moved to less green areas (n = 470) showed significantly worse mental health in the year preceding the move (P = .031) but returned to baseline in the postmove years. Moving to greener urban areas was associated with sustained mental health improvements, suggesting that environmental policies to increase urban green space may have sustainable public health benefits.


[Polypharmacy prevalence and potentially inappropriate drug prescription in the elderly hospitalized for cardiovascular disease].
Many older adults have multiple medical conditions that require medical attention, exposing them to polypharmacy and to a higher increase in prescribing potentially inappropriate medications. These situations may cause adverse drug reactions, longer hospital stays, and death. The prevalence of polypharmacy in our country is estimated at 55% and inappropriate prescribing of medications of 30%. To determine the prevalence of polypharmacy and potentially inappropriate drug prescription in older patients hospitalized for cardiovascular diseases. Patients older than 70 years from a tertiary level hospital admitted to cardiology and angiology were included, from the period 2013-2014. Fragility status, polypharmacy, and drug prescription based on the Beer's criteria were established. The results were analyzed using descriptive statistics. For inferential analysis, cross tabulations with Pearson χ2 were used. 446 patients where included, with female predominance of 56% (mean age 76.6 ± 5.9 years). The prevalence of fragility was 35.7%, polypharmacy 84.5%, and inappropriate prescription of drugs 48.9%. Coefficient correlation between inappropriate prescription of drugs and polypharmacy was p = 0.001. The prevalence of polypharmacy and inappropriate prescription of drugs was higher than reported previously, which shows the situational diagnosis in this tertiary level hospital, considering a population with high cardiovascular risk.


Global developmental delay in a 10-month-old infant boy.
A 10-month-old boy was seen for the first time for a health supervision visit by a pediatrician. A brief review of the child's medical history did not reveal any specific problems. On physical examination, the pediatrician found an alert, smiling child, but she was surprised by the following observations: unable to sit without support, absent pincer grasp, no audible language, unilateral exotropia, and microcephaly. Expansion of the medical history revealed an uneventful full-term prenatal course and normal vaginal delivery. The mother denied use of alcohol or other drugs/medications during the pregnancy. She did not have a recent history of any infections, unexplained fevers, or high risks for sexually transmitted disease. The baby cried spontaneously and the parents reported no resuscitation efforts. There were early feeding problems associated with a poor suck and gastroesophageal reflux. The parents were healthy and this was their first child. Family history was negative for early problems in child development or any neurological conditions. Parents were high school graduates without any learning problems; they were both employed in retail sales with a steady employment history. The pediatrician then took a second look at the child and discovered truncal hypotonia, extremity hypertonia, tongue protrusion, and a broad mouth. She concluded that the child had a global developmental delay, including delays in motor, language, and social development.


Long-term outcome in 44 horses with stifle lameness after arthroscopic exploration and debridement.
To (1) examine the outcome in horses with osteoarthritis or intra-articular soft tissue injuries of the stifle after arthroscopic exploration and debridement and (2) to determine any imaging or surgical findings that may influence prognosis. Case series. Horses (n=44) with lameness referable to the stifle, diagnosed with osteoarthritis, meniscal tears, or other intra-articular soft tissue injuries based on arthroscopic examination. Medical records of horses with stifle lameness that had arthroscopic exploration were reviewed. Horses with osteochondrosis lesions, intra-articular fractures, or osseous cyst-like lesions were excluded. Pertinent case information was analyzed and short- and long-term outcome was assessed. There was no association between radiographic score and surgery score. Diagnostic ultrasound had a sensitivity of 79% and a specificity of 56% for identifying meniscal injuries. Follow-up information was available for 35 horses; 23 horses (60%) improved after surgery, 16 (46%) became sound, and 13 (37%) returned to their previous level of function. A negative association was observed between age and degree of preoperative lameness and outcome. More severe changes observed on preoperative radiographs were also negatively associated with prognosis. No horses with grade 3 meniscal tears improved postoperatively and increasing meniscal pathology was negatively associated with return to previous function. A weak association between surgery grade and outcome was also observed. Degree of chondral damage, location of primary pathology, and microfracture techniques had no effect on outcome. Advanced horse age, severe lameness and preoperative radiographic changes, and presence of large meniscal tears are associated with a negative postoperative outcome for horses with stifle lameness. Appearance of the articular surface at surgery appears to be an inconsistent prognostic indicator. Some horses with extensive cartilage damage may return to athletic function after arthroscopic debridement and lavage. A more pessimistic prognosis may be given to older horses, those with more severe preoperative lameness, and those with severe radiographic changes or large meniscal tears.


New C₂₀-gibberellin diterpene from the leaves of Schefflera sessiliflora De P. V.
From the leaves of Schefflera sessiliflora De P. V., one new C20-gibberellin diterpene 2β,12β-dihydroxygibberellin (12β-hydroxy-GA110 or 2β-hydroxy-GA112) (1), together with three known compounds, trans-tiliroside (2), kaempferol 3-O-β-D-glucuronopyranoside (3), 5-p-trans-coumaroylquinic acid (4), was isolated for the first time from the genus Schefflera by various chromatography methods. Their structures were elucidated by IR, UV, HR-ESI-MS, NMR 1D and 2D experiments and comparison with previous reported data. The α-glucosidase inhibitory activity of all compounds was measured. The isolates (2, 3) showed better α-glucosidase inhibitory activity (IC50 = 134.60, 147.10 μM, respectively) than the standard drug acarbose (IC50 = 214.50 μM).


Setup of galvanic sensors for the monitoring of gilded bronzes.
Traditional electrochemical techniques, such as linear polarization resistance (Rp), and electrochemical impedance spectroscopy (EIS), cannot be applied to gilded bronzes, as it may not be possible to interpret the results obtained due to the bimetallic nature of the studied material. The measurement of the macrocouple current generated by the gold/bronze galvanic couple can be used as an indicator of degradation processes. Nevertheless, this measurement cannot be performed directly on the original artifacts due to the systematic presence of short-circuits between the two metals. In the present work the use of galvanic sensors is proposed as a possible solution for the monitoring of gilded bronze artefacts. The sensors have been designed to simulate real gilded bronze surfaces in terms of composition and stratigraphy and have proved to be a reliable diagnostic tool for the in situ monitoring of the rates of deterioration of gilded bronze surfaces and to test new conservation treatments. Their set-up and application is reported and their performances discussed.


A comparative study of psychiatric services in Japan and England.
Psychiatric services in Japan and England are compared using government statistics. In Japan, the number of in-patients per 100,000 population has increased from the 1950s, while that of England has decreased since 1954. Since 1972 the prevalence of in-patients has been higher in Japan than in England. The admission rate is lower in Japan than in England, and there are more long-stay patients. Most Japanese in-patients are admitted compulsorily, whereas most are admitted voluntarily in England. The attendance at out-patient clinics is higher in Japan than in England, but there are far fewer day-hospital places in Japan. Differing government policies are the main reason for these differences.


Assessment of serum antibody patterns and analysis of subgingival microflora of members of a family with a high prevalence of early-onset periodontitis.
In a study of members of a large family with a high prevalence of early-onset periodontitis, we sampled the subgingival microflora and characterized 40 isolates from each sample. We surveyed serum samples by enzyme-linked immunosorbent assay for antibodies reacting with any of a panel of 21 periodontal bacteria. The mother and 7 of her 13 children had early-onset periodontitis. Bacteroides gingivalis was not detected in the subgingival flora of any affected or unaffected family member, and Actinobacillus actinomycetemcomitans was isolated from only one affected child. Capnocytophaga ochracea was isolated from five of seven affected children and from none of their normal siblings. We found no significant differences among the floras from family members who had rapidly progressive, juvenile, and prepubertal forms of periodontitis. Elevated levels of serum antibody reacting with one or more of the bacteria tested were found in all family members with disease, but in only one periodontally normal family member. Both children with prepubertal periodontitis had antibodies reacting with C. sputigena, a species not found in their subgingival floras, but with none of the other bacteria tested. All remaining affected family members had antibodies to one or more serotypes of A. actinomycetemcomitans, and four had antibodies reacting with additional bacteria, including C. sputigena, Eikenella corrodens, Fusobacterium nucleatum, and Haemophilus aphrophilus. Sera from patients contained antibodies specific for putative periodontal pathogens not found in their pocket flora, and conversely, putative periodontal pathogens for which no serum antibodies were found frequently comprised a large proportion (10% or more) of the pocket flora. In no case were both the bacterium and its antibody found. These observations are suggestive of sequential infection in the early-onset forms of periodontitis and of induction of protective immunity against reinfection by the same microorganism.


Biological effectiveness of high-energy protons: target fragmentation.
High-energy protons traversing tissue produce local sources of high-linear-energy-transfer (LET) ions through nuclear fragmentation. We examine the contribution of these target fragments to the biological effectiveness of high-energy protons using the cellular track model. The effects of secondary ions are treated in terms of the production collision density using energy-dependent parameters from a high-energy fragmentation model. Calculations for mammalian cell cultures show that at high dose, at which intertrack effects become important, protons deliver damage similar to that produced by gamma rays, and with fragmentation the relative biological effectiveness (RBE) of protons increases moderately from unity. At low dose, where sublethal damage is unimportant, the contribution from target fragments dominates, causing the proton effectiveness to be very different from that of gamma rays with a strongly fluence-dependent RBE. At high energies, the nuclear fragmentation cross sections become independent of energy. This leads to a plateau in the proton single-particle-action cross section, below 1 keV/micron, since the target fragments dominate.


After how many drinks does someone experience acute consequences-determining thresholds for binge drinking based on two event-level studies.
The threshold of 4+/5+ drinks per occasion has been used for decades in alcohol research to distinguish between non-risky versus risky episodic drinking. However, no study has assessed the validity of this threshold using event-level data. This study aimed to determine the optimal thresholds for the detection of five acute alcohol-related consequences (hangover, blackout, risky sex, fights and injury) using data from two event-level studies. An event-level study to assess the ability to use the number of drinks consumed to discriminate between nights with and without consequences using the area under the receiver operating characteristic (AUROC) curve. Optimal thresholds were determined using the Youden Index based on sensitivity and specificity. Separate thresholds were estimated for gender and age groups (16-17 versus 18-25). Lausanne and Zurich, Switzerland. Three hundred and sixty-nine participants aged 16-25 years. On 3554 weekend nights, participants reported total number of alcoholic drinks consumed the previous night and acute consequences (hangover, blackout, risky sex, fights and injury) FINDINGS: Hangover was the most frequently reported consequence and injury the least for both genders. Throughout age groups and studies, optimal thresholds for any consequence, and for hangover only, were equal to 4+/5+ (40+/50+ g alcohol) while those for blackouts, risky sex, fights and injuries were up to three drinks higher. Adolescents tended to experience consequences more often and at slightly lower drinking levels than did adults. For all consequences but injuries, the optimal thresholds were one to two drinks lower for women than for men. Event-level data collection techniques appear particularly suitable to estimate thresholds at which acute alcohol-related consequences occur. Binge drinking thresholds of 4+/5+ (women/men) drinks, equivalent to 40+/50+ g pure alcohol, predict the occurrence of consequences accurately in general but are too low to predict severe acute alcohol-related consequences.


Management of osteoporosis in liver disease.
Osteoporosis resulting in a high risk for fracture is a common complication in patients with liver disease, particularly in those with chronic cholestasis and with end-stage cirrhosis. The pathogenesis of bone loss in liver patients is poorly understood but it mainly results from low bone formation as a consequence of cholestasis or the harmful effects of alcohol or iron on osteoblasts. Increased bone resorption has also been described in cholestatic women with advanced disease. The management of bone disease in liver patients is addressed to reduce or avoid the risk factors for osteoporosis and fracture. Bisphosphonates associated with supplements of calcium and vitamin D are safe and effective for increasing bone mass in patients with chronic cholestasis and after liver transplantation, though no clear achievements in descreasing the incidence of fractures have been described, probably because of the low number of patients included in the therapeutic trials. Randomized studies assessing bisphosphonates in larger series of patients, the development of new drugs for osteoporosis and the improvement in the management of liver transplant recipients may change the future.


[Cell communication in the development of the oral cavity].
The cadherins are a family of a calcium-dependent cell adhesion molecules that are regulated both spatially and temporally during development. Epithelial cadherin (E-cadherin) is present in the epithelial cells in both the embryo and yolk sac during organogenesis. In many studies, the presence of E-cadherin was analyzed during normal and disturbed craniofacial development with respect to palate and tooth formation. E-cadherin was present in epithelia of both ectodermal and endodermal origin. The expression level of E-cadherin increased with age and differentiation. The expression pattern of E-cadherin implies that this molecule has a role during normal development of the epithelia of the cranio-facial complex. Morphogenesis and cell differentiation in the developing tooth are controlled by a series of reciprocal interactions between the epithelial and mesenchymal tissues. The exact molecular mechanisms operating in these interactions are unknown at present, but both structural components of the extracellular matrix (ECM) and diffusible growth factors have been suggested to be involved. In this review article we summarize our findings on the distribution patterns of three ECM molecules and two cell surface receptors during tooth morphogenesis through bud, cap, and bell stages of development. The examined molecules include fibronectina and tenascin, which all represent components of the mesenchymal ECM, the cell surface proteoglycan, syndecan, which functions as a receptor for interstitial matrix. Based on the observed changes in distribution patterns and on experimental evidence, roles are suggested for these molecules in epithelial-mesenchymal interactions during tooth development. C-Cam is a cell surface glycoprotein that is involved in cell adhesion and may play a role in histogenesis and organogenesis. It is a member of the carcinoembryonic antigen (CEA) gene family, which is a subfamily of the immunoglobulin gene superfamily. The presence of C-CAM in all proliferating craniofacial epithelia indicates that this molecule may play an important role in development.


A Case Report of Rapidly Lethal Acute Respiratory Distress Syndrome Secondary to Coronavirus Disease 2019 Viral Pneumonia.
As of April 2020, the coronavirus 2019 (COVID-19) pandemic has resulted in more than 210,000 deaths globally. The most common cause of death from COVID-19 is acute respiratory failure. We report the case of a 78-year-old female with a history of hypertension, cerebrovascular accident (CVA), type 2 diabetes mellitus, and sarcoidosis, who presented to the emergency department with one day of dyspnea. The patient experienced a rapid decline in respiratory function and was intubated in the intensive care unit (ICU), meeting the Berlin criteria for severe acute respiratory distress syndrome (ARDS). Chest radiography revealed diffuse bilateral coalescent opacities, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA swab test was positive for COVID-19. The patient experienced acute kidney injury with uptrending creatinine levels and remained lethargic and unresponsive throughout her ICU stay, suggestive of potential hypoxic brain injury. In light of the patient's poor clinical status, age, and significant comorbidities, prognosis was conveyed about medical futility and patient's family agreed to terminal extubation and the patient expired peacefully, exactly one week from hospital admission. This case report highlights the speed at which severe ARDS can present and contribute to end-organ dysfunction in COVID-19 patients.


In vitro degradation and angiogenesis of the porous calcium silicate-gelatin composite scaffold.
Calcium silicate-based materials have attracted a great deal of interest due to their osteogenesis and have been used as implant materials for bone repair and regeneration. The purpose of this study was to use gelatin with and without genipin cross-linking for controlling degradation, improving mechanical properties, and enhancing angiogenesis of calcium silicate bioceramics. The in vitro degradation of gelatin-containing scaffolds was analysed in a simulated body fluid (SBF) solution. Human mesenchymal stem cells (hMSCs) were used to examine angiogenesis. The results indicated that the gelatin-containing scaffolds showed a diametral tensile strength of about 2 MPa and a porosity of about 60% falling within the range of values reported for the cancellous bone. Apatite precipitation occurred on all scaffold surfaces after soaking in SBF for 1 week. The gelatin-containing scaffold without cross-linking exhibited a greater weight loss and porosity than the control without gelatin. The cross-linking agent, genipin, significantly improved the mechanical stability of the composite scaffold. The gelatin enhanced the viable cell populations. More importantly, gelatin actively promoted the secretion of angiogenic factors such as von Willebrand factor and angiopoietin-1 in hMSCs. It is concluded that combination of calcium silicate and gelatin may synergistically enhance clinically desirable functions in terms of controlled degradation, improved mechanical properties, and enhanced angiogenesis.


Outcomes for patients with lung cancer admitted to intensive care units.
This study aimed to evaluate the outcomes for patients with lung cancer admitted to intensive care units and assess their clinical and demographic profiles. Retrospective, analytical, observational study, wherein the outcomes for patients diagnosed with lung cancer admitted to the intensive care unit of university hospital from January 2010 until February 2011 were evaluated. Thirty-four patients' medical records were included. Twenty-six (76.5%) patients received some type of ventilatory support, of whom 21 (61.8%) used invasive mechanical ventilation and 11 (32.4%) used noninvasive ventilation at some point during their stay at the intensive care unit. Regarding mortality, 12 (35.3%) patients died during hospitalization at the intensive care unit, totaling 15 (44.1%) deaths during the entire hospitalization period; 19 (55.9%) patients were discharged from the hospital. The analysis of the variables showed that the patients who died had remained on invasive mechanical ventilation for a longer period 5.0 (0.25 to 15.0) days than the survivors (1.0 (0 to 1.0) days) (p=0.033) and underwent dialysis during their stay at the intensive care unit (p=0.014). The mortality of patients with lung cancer admitted to the intensive care unit is associated with the time spent on invasive mechanical ventilation and the need for dialysis.


Chronic inflammatory demyelinating polyradiculoneuropathy: search for factors associated with treatment dependence or successful withdrawal.
About 40% of responders to treatment for chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) remain treatment dependent and have a relapse if treatment is interrupted. To look for factors associated with treatment dependence or successful withdrawal in CIDP patients. We retrospectively studied 70 responder CIDP patients comprising 34 patients who remained treatment dependent (treatment-dependent group) and 36 patients whose treatment could be discontinued (treatment withdrawal group). Clinical, biological, electrophysiological and therapeutic features were compared between these groups. A multifocal deficit was more frequent in the treatment-dependent group (35%) than in the treatment withdrawal group (8%) (p<0.01). The most frequent effective treatment was intravenous immunoglobulin (IVIG) for the treatment-dependent group (79%). In this group, more patients were resistant to corticosteroids in first-line therapy (93%) than in the treatment withdrawal group (40%) (p=0.002). The delay to effective treatment was significantly shorter for the treatment withdrawal group than for the treatment-dependent group (mean 11.1 vs 31.2 months; p<0.01). The rate of successful withdrawal was lower with IVIG (29%) than with corticosteroids (83%) (p<0.001). When compared with the treatment withdrawal group, the treatment-dependent group was more frequently responsive to IVIG, more frequently resistant to corticosteroids in first-line treatment, had a longer delay to effective treatment and was more likely to present a multifocal deficit. The rate of successful withdrawal seems to be higher with corticosteroids, but a prospective study with a long-term follow-up is needed to confirm these features.


State of the art and the dark side of amyotrophic lateral sclerosis.
Amyotrophic lateral sclerosis (ALS) is a disorder that involves the degeneration of motor neurons, muscle atrophy, and paralysis. In a few familiar forms of ALS, mutations in the superoxide dismutase-1 (SOD1) gene have been held responsible for the degeneration of motor neurons. Nevertheless, after the discovery of the SOD1 mutations no consensus has emerged as to which cells, tissues and pathways are primarily implicated in the pathogenic events that lead to ALS. Ubiquitous overexpression of mutant SOD1 in transgenic animals recapitulates the pathological features of ALS. However, the toxicity of mutant SOD1 is not necessarily limited to the central nervous system. Views about ALS pathogenesis are now enriched by the recent discovery of mutations in a pair of DNA/RNA-binding proteins called TDP-43 and FUS/TLS as causes of familial and sporadic forms of ALS. Although the steps that lead to the pathological state are well defined, several fundamental issues are still controversial: are the motor neurons the first direct targets of ALS; and what is the contribution of non-neuronal cells, if any, to the pathogenesis of ALS? The state of the art of ALS pathogenesis and the open questions are discussed in this review.


IL-17 and IL-22 elicited by a DNA vaccine encoding ROP13 associated with protection against Toxoplasma gondii in BALB/c mice.
Toxoplasma gondii, an intracellular parasitic protozoan, is capable of infecting man and all warm-blooded animals. Cell-mediated immunity is vital in mounting protective responses against T. gondii infection. Recent studies have shown that T-helper (Th) 17 responses may play a key role in parasite control. In this current study, we constructed a DNA vaccine encoding T. gondii ROP13 in a pcDNA vector. Groups of BALB/c mice were immunized intramuscularly with pcROP13 or controls and challenged with the RH strain of T. gondii. The results showed that immunization with pcROP13 could elicit an antibody response against T. gondii. The expression of the canonical Th17 cytokines, interleukin (IL)-17 and IL-22, were significantly increased after immunization with pcROP13 compared with control groups ( p < 0.05). Furthermore, vaccination resulted in a significant decrease in parasite load ( p < 0.05). The induction of Th17 related cytokines, using a ROP13 DNA vaccine, against T. gondii should be considered as a potential vaccine approach for the control of toxoplasmosis.


Do laboratory species protect endangered species? Interspecies variation in responses to 17β-estradiol, a model endocrine active compound.
Although the effects of estrogens on model laboratory species are well documented, their utility as surrogates for other species, including those listed as endangered, are less clear. Traditionally, conservation policies are evaluated based on model organism responses but are intended to protect all species in an environment. We tested the hypothesis that the endangered Rio Grande silvery minnow (Hybognathus amarus) is more vulnerable to endocrine disruption-as assessed through its larval predator-escape performance, survival, juvenile sex ratios, and whole-body vitellogenin concentration-than the commonly used toxicological model species fathead minnow (Pimephales promelas) and the bluegill sunfish (Lepomis macrochirus). Fish were exposed concurrently for 21 days to the model endocrine active compound (EAC) 17ß-estradiol (E2) at 10 ng E2/L and 30 ng E2/L in a flow-through system using reconstituted water that simulated the physicochemical conditions of the Middle Rio Grande in New Mexico, USA. No significant differences were observed between the fathead and silvery minnow in larval predator-escape response or juvenile sex ratio. Rio Grande silvery minnow survival decreased significantly at day 14 compared with the other two species; by day 21, both cyprinid species (silvery minnow and fathead minnow) exhibited a significant decrease in survival compared with bluegill sunfish, a member of the family Centrarchidae. Male Rio Grande silvery minnow showed a significant increase in whole-body vitellogenin concentration in the 10 ng/L treatment, whereas fathead minnow and bluegill sunfish showed no significant increases in vitellogenin concentrations across treatments. Our study showed response differences to estrogen exposures between the two cyprinid species and further divergence in responses between the families Cyprinidae and Centrarchidae. These results suggest that commonly used laboratory model organisms may be less sensitive to EACs than the endangered Rio Grande silvery minnow. However, this study supports the continued use of surrogate species for the beneficial implementation of water-quality regulations for the protection of threatened and endangered species if phylogenetic relationships are taken into consideration.


Role of the 25 kDa major outer membrane protein of Legionella pneumophila in attachment to U-937 cells and its potential as a virulence factor for chick embryos.
The gene encoding the 25 kDa major outer membrane protein (MOMP) of Legionella pneumophila was transformed into Escherichia coli JM 83 and the resultant E. coli LP 116 clone expressed the Legionella-MOMP. Compared with the parent E. coli strain, the clone showed a fivefold increase in opsonin-independent binding to U-937 cells. Furthermore, this gene was incorporated by electroporation into a low virulence derivative of Leg. pneumophila which showed reduced expression of the MOMP but enhanced expression of a 31 kDa protein in the OMP profile. After electroporation, the attenuated strain showed an increased expression of the MOMP while the 31 kDa protein was eliminated and virulence for the chick embryo was re-established. The use of a monoclonal antibody specific for the MOMP abolished virulence and adherence. These studies suggest that the 25 kDa MOMP of Leg. pneumophila serves as an adhesive molecule for host cells and that this protein plays a major role in the virulence of the organism for the chick embryo.


Electromyographic analysis of walking in water in healthy humans.
This study was designed to describe and clarify muscle activities which occur while walking in water. Surface electromyography (EMG) was used to evaluate muscle activities in six healthy subjects (mean age, 23.3 +/- 1.4 years) while they walked on a treadmill in water (with or without a water current) immersed to the level of the xiphoid process, and while they walked on a treadmill on dry land. The trials in water utilized the Flowmill which has a treadmill at the base of a water flume. Integrated EMG analysis was conducted for the quantification of muscle activities. In order to calculate the %MVC, the measurement of maximal voluntary contraction (MVC) of each muscle was made before the gait analysis, thus facilitating a comparison of muscle activities while walking in water with those on dry land. The %MVCs obtained from each of the tested muscles while walking in water, both with and without a water current, were all found to be lower than those obtained while walking on dry land at a level of heart rate response similar to that used when walking on dry land. Furthermore, the %MVCs while walking in water with a water current tended to be greater when compared to those while walking in water without a water current. In conclusion, the present study demonstrated that muscle activities while walking in water were significantly decreased when compared to muscle activities while walking on dry land, that muscle activities while walking in water tended to be greater with a water current than without, and that the magnitude of the muscle activity in water was relatively small in healthy humans. This information is important to design water-based exercise programs that can be safely applied for rehabilitative and recreational purposes.


Population genomic analysis of outcrossing and recombination in yeast.
The budding yeast Saccharomyces cerevisiae has been used by humans for millennia to make wine, beer and bread. More recently, it became a key model organism for studies of eukaryotic biology and for genomic analysis. However, relatively little is known about the natural lifestyle and population genetics of yeast. One major question is whether genetically diverse yeast strains mate and recombine in the wild. We developed a method to infer the evolutionary history of a species from genome sequences of multiple individuals and applied it to whole-genome sequence data from three strains of Saccharomyces cerevisiae and the sister species Saccharomyces paradoxus. We observed a pattern of sequence variation among yeast strains in which ancestral recombination events lead to a mosaic of segments with shared genealogy. Based on sequence divergence and the inferred median size of shared segments (approximately 2,000 bp), we estimated that although any two strains have undergone approximately 16 million cell divisions since their last common ancestor, only 314 outcrossing events have occurred during this time (roughly one every 50,000 divisions). Local correlations in polymorphism rates indicate that linkage disequilibrium in yeast should extend over kilobases. Our results provide the initial foundation for population studies of association between genotype and phenotype in S. cerevisiae.