Mutations that prevent or mimic persistent post-translational modifications of the histone H3 globular domain cause lethality and growth defects in Drosophila.
Understanding the function of histone post-translational modifications is the key to deciphering how genomic activities are regulated. Among the least well-understood histone modifications in vivo are those that occur on the surface of the globular domain of histones, despite their causing the most profound structural alterations of the nucleosome in vitro. We utilized a Drosophila system to replace the canonical histone genes with mutated histone transgenes. Mutations predicted to mimic or prevent acetylation on histone H3 lysine (K) 56, K115, K122, and both K115/K122, or to prevent or mimic phosphorylation on H3 threonine (T) 118 and T80, all caused lethality, with the exception of K122R mutants. T118 mutations caused profound growth defects within wing discs, while K115R, K115Q, K56Q, and the K115/K122 mutations caused more subtle growth defects. The H3 K56R and H3 K122R mutations caused no defects in growth, differentiation, or transcription within imaginal discs, indicating that H3 K56 acetylation and K122 acetylation are dispensable for these functions. In agreement, we found the antibody to H3 K122Ac, which was previously used to imply a role for H3 K122Ac in transcription in metazoans, to be non-specific in vivo. Our data suggest that chromatin structural perturbations caused by acetylation of K56, K115, or K122 and phosphorylation of T80 or T118 are important for key developmental processes.


Erratum: Radiolabeling and Preliminary Evaluation of 99mTc-Labeled DNA Cube Nanoparticles as Potential Tracers for SPECT Imaging [Corrigendum].
[This corrects the article DOI: 10.2147/IJN.S325791.].


Hyaluronic acid and hyaluronidase activity in gingival exudate from sites of acute ulcerative gingivitis in man.
Gingival exudates from sites of acute ulcerative gingivitis (AUG) and chronic gingivitis (CG) in adults were investigated by cellulose-acetate electrophoresis for the hyaluronic acid (HA) content and assayed for the levels of HA-degrading enzymes. HA was the only glycosaminoglycan (GAG) in gingival exudate from CG sites. HA was not detected at untreated AUG sites but was evident, at increasing levels, after two and seven days of effective antibacterial treatment. In AUG exudates, the total HA-degrading enzyme activity, of bacterial origin, decreased to approx. 30 and 10 per cent of the high initial levels after two and seven days of treatment respectively, to that level found at sites of CG. The specific activity of HA-degrading enzyme of lysosomal origin was low initially and increased with treatment to a level comparable to CG. The notable absence of HA from gingival exudate from sites of untreated AUG thus appears to result from the increased levels of bacterial hyaluronidase. Electrophoresis of gingival exudate may be an indirect method of monitoring the rate of response of AUG to different antibacterial treatments.


Correction to "Red blood cell distribution width as a novel marker for predicting bleeding after endoscopic resection for early gastric cancer".
[This corrects the article DOI: 10.1002/deo2.123.].


Sorption of atrazine and phenanthrene by organic matter fractions in soil and sediment.
Atrazine and phenanthrene (Phen) sorption by nonhydrolyzable carbon (NHC), black carbon (BC), humic acid (HA) and whole sediment and soil samples was examined. Atrazine sorption isotherms were nearly linear. The single-point organic carbon (OC)-normalized distribution coefficients (K(OC)) of atrazine for the isolated HA1, NHC1 and BC1 from sediment 1 (ST1) were 36, 550, and 1470 times greater than that of ST1, respectively, indicating the importance of sediment organic matter, particularly the condensed fractions (NHC and BC). Similar sorption capacity of atrazine and Phen by NHC but different isotherm nonlinearity indicated different sorption domains due to their different structure and hydrophobicity. The positive relationship between (O+N)/C ratios of NHC and atrazine logK(OC) at low concentration suggests H-bonding interactions. This study shows that sediment is probably a less effective sorbent for atrazine than Phen, implying that atrazine applied in sediments or soils may be likely to leach into groundwater.


Spontaneous conversion of permanent atrial fibrillation into stable sinus rhythm after 17 months of biventricular pacing.
A 68-year-old woman with drug refractory congestive heart failure (NYHA Class III, ejection fraction 35%, moderate to severe mitral regurgitation) and permanent atrial fibrillation uninterrupted for >3 years, underwent implantation of a biventricular pacing system. Seventeen months later, atrial fibrillation spontaneously converted into sinus rhythm, which has been maintained for more than 6 months (to date).


An evaluation of the notification of birth form.
Data on 108 Notification of Birth Forms, from a total of 6 hospitals, have been compared with hospital case notes, for both accuracy and completeness. Items related to socio-demographic features of mothers were reliable. However, certain items related to past obstetric history, such as date of last birth and number of previous live births, were less than 90% reliable. Data on index pregnancy were poorest, with three items, first visit to doctor during pregnancy, first visit to hospital during pregnancy and rubella immune status all less than 75% reliable. Data from one hospital were of poorer quality than others, principally because the information on the form is not obtained directly from the mother. Data on forms related to travellers (N = 29) were also of poorer quality.


Oral squamous cell carcinoma; from an hypothesis about a virus, to concern about possible sexual transmission.
Nearly two decades ago, we produced the first evidence for the presence of viral nucleic acids in oral squamous cell carcinoma (OSCC) tissues, hypothesising that there may be a viral involvement in at least some OSCC. Subsequently, human papillomaviruses (HPV) in particular have been implicated in OSCC. Antibody responses to HPV are seen and HPV-DNA detected in tumors by us and many others, the virus being mainly HPV-16, the genotype associated with ano-genital cancer. HPV are seen by in situ hybridisation only in tumour and premalignant tissue but not in surrounding normal mucosa suggesting HPV has a causal relationship. HPV may also be integrated in the host genome, further suggesting a causal role. Studies of patients with OSCC have suggested possible sexual transmission of HPV. Recent studies have indicated that HPV may be aetiologically important particularly in some types of oropharyngeal cancer, at least in tonsillar carcinogenesis, and may represent an alternative pathway in carcinogenesis to the established factors of tobacco and alcohol. We have come a very long way in the two decades since our first suggestion of a viral aetiopathogenesis was greeted with incredulity, and data from on-going studies by the International Agency for Research on Cancer, Johns Hopkins Oncology Center and others are eagerly awaited.


Regulation of surfactant phospholipid secretion from isolated rat alveolar type II cells by lectins.
The major surfactant-associated protein is a potent inhibitor of surfactant phospholipid secretion from isolated type II cells. Since the major surfactant-associated protein contains a carboxy terminal polypeptide domain which is homologous to the lectin-like liver mannose-binding protein, we tested whether lectins inhibit surfactant phospholipid secretion from rat alveolar type II cells. Concanavalin A, wheat germ agglutinin and Maclura pomifera agglutinin were potent inhibitors of surfactant phospholipid secretion. When adenosine 5'-triphosphate (ATP) was utilized as a secretagogue, the IC50 values for inhibition of surfactant phospholipid secretion were 5.10(-7) (wheat germ agglutinin), 1.10(-6) (concanavalin A) and 2.5.10(-5) M (M. pomifera agglutinin). Similar results were obtained when 12-O-tetradecanoylphorbol 13-acetate was utilized as a secretagogue: IC50 values of 1.10(-6) M for concanavalin A and wheat germ agglutinin and 2.5.10(-5) M for M. pomifera agglutinin. Hapten sugars were utilized to antagonize the inhibitory effect of the lectins. N-Acetyl-D-glucosamine significantly reversed inhibition of phospholipid secretion by wheat germ agglutinin in a dose-dependent fashion and methyl alpha-D-mannoside significantly reversed inhibition of phospholipid secretion by concanavalin A. N-Acetyl-D-galactosamine had no significant effect on inhibition of secretion produced by any of the lectins. The inhibitory effect of the lectins did not appear to be due to cytotoxicity since lactate dehydrogenase was not released above control levels and the inhibition of the surfactant phospholipid secretion by wheat germ agglutinin could be reversed after treatment of cells with wheat germ agglutinin by washing the lectin from the cells followed by treatment of the cells with ATP. These studies demonstrate a direct inhibitory effect of plant lectins on phospholipid secretion from type II cells in vitro.


Drug response patterns as a basis of nosology for the mood-incongruent psychoses (the schizophrenias).
Interaction of therapeutic drugs with a series of different biopathological substrates of psychosis might be expected to generate a series of different response patterns. Herein the authors suggest that multi-modal response patterns following lithium and neuroleptic treatment of psychotic patients may aid in resolving the heterogeneity of psychotic disorders and lead to a new nosology of the psychoses.


Extensive jejunal injury is repaired by migration and transdifferentiation of ileal enterocytes in zebrafish.
A major cause of intestinal failure (IF) is intestinal epithelium necrosis and massive loss of enterocytes, especially in the jejunum, the major intestinal segment in charge of nutrient absorption. However, mechanisms underlying jejunal epithelial regeneration after extensive loss of enterocytes remain elusive. Here, we apply a genetic ablation system to induce extensive damage to jejunal enterocytes in zebrafish, mimicking the jejunal epithelium necrosis that causes IF. In response to injury, proliferation and filopodia/lamellipodia drive anterior migration of the ileal enterocytes into the injured jejunum. The migrated fabp6+ ileal enterocytes transdifferentiate into fabp2+ jejunal enterocytes to fulfill the regeneration, consisting of dedifferentiation to precursor status followed by redifferentiation. The dedifferentiation is activated by the IL1β-NFκB axis, whose agonist promotes regeneration. Extensive jejunal epithelial damage is repaired by the migration and transdifferentiation of ileal enterocytes, revealing an intersegmental migration mechanism of intestinal regeneration and providing potential therapeutic targets for IF caused by jejunal epithelium necrosis.


COVID-19, Social Determinants Past, Present, and Future, and African Americans' Health.
As the COVID-19 pandemic progresses, more African Americans than whites are falling ill and dying from the virus and more are losing livelihoods from the accompanying recession. The virus thereby exploits structural disadvantages, rooted partly in historical and contemporary anti-Black sentiments, working against African Americans. These include higher rates of comorbid illness and more limited health care access, higher rates of disadvantageous labor market positioning and community and housing conditions, greater exposure to long-term care residence, and higher incarceration rates. COVID-19 also exposes African Americans' greater vulnerability to recession, and possibly greater susceptibility to accompanying behavioral health problems. If they are left unaddressed, the very vulnerabilities COVID-19 exploits may perpetuate themselves. However, continuing and supplementing health and economic COVID mitigation policies can disproportionately benefit African Americans and reduce short- and long-term adverse effects. The greater impact of COVID-19 on African Americans demonstrates the consequences of pervasive social and economic inequality and marks this as a critical time to prevent further compounding of adverse effects.


Assessment of the impact of the biological larvicide VectoMax G: Combination of Bacillus thuringiensis and Lysinibacillus sphaericus on non-target aquatic organisms in Yaoundé-Cameroon.
There has been a renewed interest for larviciding during the recent decade. Although biological larvicides are considered not to be harmful to non-target organisms, there is still not sufficient data on the effect of new long-lasting larvicide formulations such as VectoMax G combining Bacillus thuringiensis israelensis and Lysinibacullus sphaericus on the environment especially on non-target organisms. The present study aimed to assess the possible influence of VectoMax G on the diversity and abundance of the aquatic fauna cohabiting with mosquito larvae in breeding habitats during a larviciding trial in the city of Yaoundé. Twelve districts of the city of Yaoundé divided into 6 intervention and 6 control sites were chosen for the study. In each district 4 semi-permanent or permanent aquatic habitats were followed. VectoMax G application was done once every two weeks during 6 months and aquatic organisms were collected 48 h after each treatment. All collected organisms were brought to the laboratory for identification. Physico-chemical parameters were recorded as well. A high diversity of the zooplankton was recorded in the intervention areas with 28 species collected against 14 species in the control areas. Cladocerans were the most represented group in both sites while Ostracods were found only in control sites. A total of 19 macro-invertebrates species were recorded in the control areas vs 16 species in the intervention areas. Gasteropods were the most represented groups of macro-invertebrates. Vertebrates such as larvivorous fishes and amphibians larvae were also found in approximately similar densities in both sites. The study indicated no significant influence of larviciding with VectoMax G on the diversity and abundance of the non-target aquatic fauna in the city of Yaoundé.


[Zinc and vitamin A deficiency in diseases of the mouth mucosa].
The regeneration of the oral mucosa is zinc- and vitamin A-pathway dependent. Hyperkeratoses, a disturbed immune system and a higher incidence of oral malignancies have been reported in cases of deficiency. In this study, blood samples were taken from 97 patients with oral lichen planus, oral leukoplakia, oral squamous cell carcinoma (OSCC), and a control group. Besides routine blood analysis, serum levels of zinc (Zn), vitamin A (Vit A) and retinol binding protein (RBP) were identified. Mean values in lichen ruber planus were Zn 784 micrograms/l, RBP 5.26 mg/dl and Vit A 656 ng/ml; in leukoplakia Zn 696 micrograms/l, RBP 5.30 mg/dl and Vit A 652 ng/ml; and in OSCC Zn 659 micrograms/l, RBP 5.00 mg/dl and Vit A 617 ng/ml. Patients with lichen ruber planus showed no statistically significant differences, whereas in leukoplakia patients there were statistically lower values for Zn compared to the control group and the lichen ruber planus group (P < 0.05). Patients with oral malignancies showed lower serum levels for Zn, RBP and Vit A. Compared with the control group and the lichen ruber planus group, differences of the serum Zn levels proved to be statistically significant (P < 0.01). In patients with oral leukoplakia and patients with oral carcinoma and Zn deficiency, a substitutional therapy should be considered.


Residential mobility during pregnancy: implications for environmental teratogenesis.
Although most teratogens are suspected to act early in the first trimester of pregnancy, birth defects monitoring programs and etiologic studies usually use residence at birth as a proxy measure for residence in the first trimester in searching for environmental teratogens. Because of the high mobility of the U.S. population, residence misclassification can potentially alter inferences concerning environmental teratogens. To evaluate this potential bias, data from the population-based Maryland Birth Defects Reporting and Information System were analyzed. In 1984, the system ascertained 295 infants with one or more of 12 sentinel defects. Of these cases, 59 (20%) mothers reported they have changed address between the time of conception and the time of birth, and 22 have moved to a different county. The residential mobility rate varied by demographic variables and was highest among white women, in the age group 20-24 years. If residence at birth is used as a screening test for residence at conception, it can be shown that in the presence of an environmental teratogenic exposure, misclassification of exposure increases with increasing mobility rate, and population exposure frequency. Such misclassification tends to weaken associations between residence and birth defects and may lead to missing environmental teratogens. This analysis emphasizes the need to use residence information early in pregnancy rather than exclusively at birth.


Toxaphene and other persistent organochlorine pesticides in three species of albatrosses from the north and south Pacific Ocean.
Toxaphene and other persistent organochlorine (OC) pesticides (chlordane-related compounds [sigmaCHL], DDT-related compounds [sigmaDDT], hexachlorocyclohexanes [sigmaHCH], tris(p-chloro-phenyl)methane, hexachlorobenzene, octachlorostyrene, dieldrin) were determined in fat of Laysan albatross (Diomedea immutabilis) and in fat and eggs of blackfooted albatross (Diomedea nigripes) from the central north Pacific Ocean. The HCH isomers and chlordane- and DDT-related compounds were also determined in eggs of northern royal albatross (Diomedea sanfordi) collected in New Zealand. Toxaphene was detected in fat samples at mean +/- standard deviation (SD) levels ranging from 243 +/- 61 ng/g wet weight in Laysan albatross to 1,020 +/- 237 ng/g wet weight in blackfooted albatross. These levels were higher than sigmaCHL and sigmaHCH but lower than sigmaDDT. In eggs of blackfooted albatross, toxaphene was the major OC pesticide, averaging 513 ng/g wet weight in two pooled samples compared with 293 ng/g wet weight for sigmaDDT. Two toxaphene congeners, the octachloroborane B8-1413 (Parlar 26) and the nonachlorobornane B9-1679 (P50), comprised about 38% of total toxaphene in both albatross species. All OC compounds were present at significantly higher levels in blackfooted than Laysan albatross fat with the exception of sigmaHCH, dieldrin, and octachlorostyrene. Mean levels of sigmaDDT and sigmaHCH in northern royal albatross eggs from New Zealand were 4 and 60 times lower, respectively, than in blackfooted albatross eggs. The pattern of OC pesticide accumulation was consistent with differences in distribution of the three species in the Pacific Ocean, with highest levels in blackfooted albatross, which feed off the west coast of North America, intermediate levels in Laysan albatross, which frequent the western Pacific, and lowest levels in northern royal albatross, which are confined to the southern oceans surrounding the Antarctic.


The PROB-PD trial: a pilot, randomised, placebo-controlled study protocol to evaluate the feasibility and potential efficacy of probiotics in modulating peripheral immunity in subjects with Parkinson's disease.
Parkinson's disease (PD) is a common neurodegenerative disease. No disease-modifying treatment is available, and therapy is symptomatic. The histopathologic hallmark is the loss of dopaminergic neurons and accumulation of α-synuclein (α-syn) in surviving neurons, but the underlying pathophysiology is unclear. Inflammatory mechanisms seem to play a prominent role, with an imbalance of immune functions and neurotoxicity caused by reactive oxygen species (ROS). Involvement of peripheral adaptive immunity, with an imbalance in T cell subpopulations and in the expression of transcriptional factors in CD4+ T cells, has also been reported. Although clinical presentation is defined by motor symptoms, patients also report non-motor symptoms, often before the onset of a clinically established disease. Etiopathogenesis of PD is unknown, but an initial aggregation of α-syn in the gut, with subsequent propagation along the vagus nerve to the brain has been hypothesised. Interestingly, in an α-syn overexpressing murine model, the absence of gut microbiota prevented both microglia activation and motor impairment, thus pointing to a fundamental role of microbiota in the development of PD. Magistrelli et al. showed that in peripheral blood mononuclear cells of PD patients, probiotics modulate the in vitro production of cytokines toward an anti-inflammatory profile and reduce the production of ROS. This is a pilot randomised placebo-controlled clinical trial protocol for a 12-week treatment with probiotics. At least 80 patients affected by PD will be recruited and randomly allocated to either the treatment or placebo group in a 1:1 ratio. General inclusion criteria will be the onset of PD 2 to 5 years before the trial and absence of autoimmune comorbidities or immunomodulating therapy. Our primary endpoint is the assessment of changes in extracellular cytokine levels (Interferon (IFN)-γ, tumour necrosis factor (TNF)-α, interleukin (IL)-4, and IL-10) and ROS production. Secondary outcomes include changes in lymphocyte subpopulations and transcriptional factors mRNA levels. This study is designed to highlight the potential beneficial role of probiotics administration on peripheral immunity through the modulation of gut microbiota. Explorative outcomes will be evaluated to assess variations in motor and non-motor symptoms and the possible correlation with probiotics administration. ClinicalTrials.gov ID NCT05173701. Registered 08 November 2021.


Beat the Blues decreases depression in financially strained older African-American adults.
Financial strain, defined as inadequate income to cover basic needs, is related to depression in minority populations. It is unclear whether interventions can improve depression in those reporting financial strain. Randomized controlled trial. Philadelphia senior center. A total of 208 African-American adults 55 years of age and older with mild to severe depressive symptoms. Beat the Blues (BTB) depression intervention, which includes care management, referral/linkage, stress reduction, depression education, and behavioral activation. Measurements included responses to the Patient Health Questionnaire-9, financial strain, and age. Although financial strain was related to depressive symptoms at baseline, participants receiving BTB at all levels of financial strain showed reduced depressive symptoms compared with a waitlist control group at 4 months. BTB participants reporting financial strain had a mean (standard deviation) decrease of 6.4 (0.85) points on the Patient Health Questionnaire-9, reflecting a clinically meaningful improvement. This nonpharmacologic intervention reduced depressive symptoms in both African-American subjects with and without financial strain. However, screening for and helping people manage financial strain may boost the magnitude of intervention benefit for those with financial strain and should be explored as part of depression care.


In crystallo observation of active site dynamics and transient metal ion binding within DNA polymerases.
DNA polymerases are the enzymatic catalysts that synthesize DNA during DNA replication and repair. Kinetic studies and x-ray crystallography have uncovered the overall kinetic pathway and led to a two-metal-ion dependent catalytic mechanism. Diffusion-based time-resolved crystallography has permitted the visualization of the catalytic reaction at atomic resolution and made it possible to capture transient events and metal ion binding that have eluded static polymerase structures. This review discusses past static structures and recent time-resolved structures that emphasize the crucial importance of primer alignment and different metal ions binding during catalysis and substrate discrimination.


Disease modification in chronic obstructive pulmonary disease.
Chronic obstructive pulmonary disease (COPD) is a preventable and treatable disease in which patients experience a progressive decline in lung function, worsening exercise capacity, and frequent exacerbations. Based on clinical evidence, the progression of COPD could be modified by focus on earlier diagnosis; risk reduction through smoking cessation; symptom reduction with pharmacotherapy, improving health-related quality of life, and pulmonary rehabilitation; and decreasing complications by reducing exacerbations. Smoking cessation has been shown to slow lung function decline and reduce mortality, including deaths due to cardiovascular disease, lung cancer, and other respiratory disease (including COPD).


Lacrimal gland as the major source of mouse tear factors that are cytotoxic to corneal keratocytes.
We have previously shown that mouse tears are cytotoxic to the corneal keratocytes. Since tear components are derived from both lacrimal tissues and ocular surface epithelium, we sought to determine the source of the cytotoxic factors in the mouse tear fluid. Cytotoxicity to keratocytes was assessed by an ex vivo assay using an isolated eye; after treatment with test samples, segmentation and disappearance of stromal nuclei were determined by DAPI nuclear staining. Following biological tissues and fluids were examined either directly or after preincubation at 37 degrees C for 2-15 hr: extraorbital lacrimal gland (ELG), intraorbital lacrimal gland (ILG), Harderian gland, Meibomian gland, corneal epithelium, bulbar conjunctiva, palpebral conjunctiva, serum, aqueous humor, and lacrimal fluid collected from a secretory duct of ELG. Under the ex vivo assay conditions, ELG and ILG, with or without preincubation, exhibited a cytotoxic effect comparable to that of diluted tears. Lacrimal fluid collected from an ELG duct was similarly effective. These specimens triggered nuclear segmentation that is typical of apoptotic nuclei. All other specimens showed no effect on stromal nuclei under the identical conditions. In some animals, ELG was surgically removed and the tear cytotoxicity was examined in vivo. The tear cytotoxicity in these animals was lost after the surgery, indicating an involvement of ELG, but it was restored 4-24hr afterward, suggesting a compensatory role of ILG.These results suggest that ELG and ILG are the major sources of tear factors that are cytotoxic to the keratocytes in the mouse.


Radiocesium (137Cs) accumulation by fish within a legacy reactor cooling canal system on the Savannah River Site.
The aquatic cooling canal system associated with a nuclear reactor built in the early 1950s received accidental releases of radiocesium (137Cs) from the reactor between 1954 and 1964, resulting in the dispersion of ~8.2 × 1012 Bq of 137Cs into the associated canals and ponds. The primary purpose of this study was to document concentrations of 137Cs in littoral zone fish currently occupying components of the cooling canal system, 3 canals and 2 impoundments, to determine how concentrations varied among these various components. Secondarily, we examined for potential influence of weirs within the canal system on concentrations in fish as well as the potential relationship between fish species and body size and on 137Cs concentrations in fish. We collected samples of sediment, biofilm, and fish from each component of the R-Reactor cooling system and compared 137Cs among sites and species in individual sites. Concentrations of 137Cs in sediments, biofilms and mosquitofish varied significantly among sampling areas with higher concentrations in RCAN1, a canal segment that was the closest to the reactor and received reactor effluent for a longer period than other components. Comparisons among other components of the cooling system, and species comparisons relative to presumed trophic positions and fish length were not consistent. However, littoral zone fish in the cooling canal system continue to bioaccumulate 137Cs >50 years after the original releases of contamination.


Effects of short-term freezing on nitrous oxide emissions and enzyme activities in a grazed pasture soil after bovine-urine application.
Nitrous oxide (N2O) emissions from soils applied with livestock excreta have been widely reported previously. The highest N2O emissions from soils are also often reported during thawing periods in cold regions where soil freezing is common. However, the combined effects of cow urine application and freeze-thaw events on N2O emissions and the related enzyme activities are still not clear. Thus, we simulated a freeze-thaw event at -3 °C for 7 days, and then increased to 3 °C for 46 days using intact soil cores with cow urine (392 kg N ha-1). We compared the factors influencing the magnitudes of N2O emissions through soil microbial processes with and without the freeze-thaw event. Dicyandiamide (DCD), an inhibitor of nitrification, was added to investigate the significance of nitrification on N2O emissions. The N2O emission rates from the urine-applied soils peaked to approximately 1000 μg N2O-N m-2 h-1 immediately after the soils thawed. Soil freezing with urine application was significantly higher cumulative N2O emissions (537 mg N2O-N m-2), compared to non-frozen soils with urine (247 mg N2O-N m-2) during the incubation period (54 days). The effect of DCD application on N2O emissions was not clear during the freeze-thaw event, although nitrate production rates were reduced. After the freezing event, soil moisture (water-filled pore space) was significantly higher in the non-frozen soils compared to the frozen soils, due to a 9% decline in bulk density of frozen soils. Additionally, the impact of thawing on urease and denitrification enzyme activities was influenced by the urine application. Urine application increased the urease activity, while the freezing event decreased the magnitudes. The physical changes in the soils were also important controlling factors of the N2O emissions from cow urine-applied soils in cold regions.


Determination of gibberellins in fermentation broth produced by Fusarium verticilliodes MTCC 156 by high-performance liquid chromatography tandem mass spectrometry.
A method for the detection of gibberellins produced by Fusarium verticilliodes is described using HPLCMS/MS (HPLC tandem MS). A Hypersil (5 microm) octadecylsilane column with methanol/water as eluent in the ratio 3:1 at a flow rate of 0.5 ml/min was used. In the HPLCMS, GA(3) (gibberellic acid; m / z 346.3) eluted at retention time tr=3.08 min, with the corresponding mass chromatogram having peaks at m / z 346.7 and 328.8 corresponding to the M+ and M+-H2O ions respectively. The ethyl acetate extract from the broth, subjected to HPLCMS analysis under similar conditions, showed a constituent with tr=2.13 min, the mass chromatogram of which exhibited peaks at m / z 348.9 and 331.9 corresponding to the MH+ and MH+-H2O ions respectively. Comparison of the MS and MS/MS results (direct infusion) of an authentic sample of GA3 and the ethyl acetate extract from the broth revealed the formation of reduced GA3 in the broth. The present study, utilizing HPLCMS/MS, describes an improved methodology for the unambiguous determination and estimation of gibberellins from fermentation broth.


On the efficacy of detoxifying agents in the prevention of fusariotoxicoses - A critical evaluation of the situation.
Fusarium toxins are of great practical relevance in animal feeding since they may occur in toxicologically relevant concentrations. Therefore, many attempts have been made to search for detoxification of contaminated feedstuffs or diets in order to cope with the problem. The supplementation of contaminated diets with detoxifying agents seems to be most feasible, andin vitro results seem to be convincing.According to the Guideline 87/153/EEC, the efficacy has to be proven by using an experimental design justified according to the claim for the use of the additive. Reviewing the literature, only a few studies investigated specific parameters which may clearly reflect the claimed mode of action of the additives, and those demonstrated no measurable detoxifying effects. The majority of investigations focused on the rather non-specific performance parameters, while many of these applied incomplete experimental designs. Nevertheless, most of the experiments did not demonstrate preventative effects.It is concluded that testing of currently available detoxifying agents did not follow the Council Directive since the claim for their use was not proven.The application of complete two by two factorial experimental designs, the investigation of mycotoxins and/or metabolites in physiological samples as specific parameters and the verification of the specificity of the detoxifying agent is recommended for futurein vivo investigations.


Mpg1, a fission yeast protein required for proper septum structure, is involved in cell cycle progression through cell-size checkpoint.
Using a yeast two-hybrid screen we isolated a gene from Schizosaccharomyces pombe which corresponds to the previously uncharacterized ORF SPCC1906.01. We have designated this gene as mpg1, based on the putative function of its product as a mannose-1-phosphatase guanyltransferase. Mpg1 shows strong similarity to other GDP-mannose-1-phosphate guanyltransferases involved in the maintenance of cell wall integrity and/or glycosylation. This homology, together with the protein's localization pattern demonstrated in this work, strongly suggests that Mpg1 is involved in cell wall and septum synthesis. Moreover, cells lacking Mpg1 present a defect in glycosylation, are more sensitive to Lyticase, and show an aberrant septum structure from the start of its deposition, indicating that the Mpg1 function is necessary for the correct assembly of the septum. Interestingly, lack of Mpg1 clearly affects cell cycle progression: mpg1 null mutants arrest as septated and bi-nucleated 4C cells, without an actomyosin ring. Wee1 is required for the G2/M arrest induced in the absence of Mpg1, since the blockade is circumvented when Wee1 is inactivated. Wee1 is part of a cell-size checkpoint that prevents entry into mitosis before cells reach a critical size. The results presented in this work demonstrate that the G2/M arrest induced in the absence of Mpg1 is mediated by this cell size checkpoint, since oversized mutant cells enter mitosis. The mpg1 loss-of-function mutant, therefore, provides a good model in which to study how cells coordinate cell growth and cell division.


Interaction of DNA with aromatic hydrocarbons fraction in atmospheric particulates of Xigu District of Lanzhou, China.
Voluminously epidemiological studies show that the relationships exist between the air pollution and human health and cancer. Aromatic hydrocarbons (AHs) in air form a large class of organic pollutants, which are widely in environment and many of them are known to be carcinogenic and/or mutagenic and contribute to ambient air pollution. In the past decades, bioassays mainly have been used to evaluate the toxicity of chemical mixtures in atmospheric particulates or aqueous environment. However, it is well known that the covalent complexes formed by carcinogens with DNA may be exert negative results in bioassay. So the main aim of this paper is to develop an evaluation method of toxicity effects of chemical mixtures in atmospheric particulates from chemical standpoint. In this study, the in vitro interaction of the AHs with DNA was investigated by absorption, fluorescence and resonance light scattering (RLS) spectroscopic techniques. The results showed that the AHs in the atmospheric particulates could combine with calf thymus DNA (ctDNA) and herring sperm DNA (hsDNA) without being activated or metabolized by organism, respectively. Intercalation may be present in the mechanism of interaction. The binding constants of the AHs with ctDNA and hsDNA were 2.5 x 10(2) and 2.0 x 10(3), respectively, which indicated that the interaction of the AHs with hsDNA is stronger than that with ctDNA. In addition, the relationships of dose-effect between the total mole concentration of chemical components and the ability of binding ctDNA and hsDNA were confirmed. This research made it possible to study the toxicity effects of chemical mixtures in atmospheric particulates by chemical method. It is believed that the composition and contents of unknown AHs and the interaction of DNA with AHs in atmospheric particulates of Xigu District of Lanzhou City, China are first reported in the past twenty years.


Cricoid pressure during intubation: A systematic review and meta-analysis of randomised controlled trials.
This systematic review and meta-analysis aimed to determine whether cricoid pressure protects against aspiration and whether this technique adversely affects intubating conditions in adult patients. A systematic review of five databases was performed for randomised controlled trials comparing cricoid pressure to no cricoid or sham cricoid during intubation. The primary outcome was incidence of aspiration and the secondary outcomes included first attempt intubation success, time to intubation, Cormack and Lehane Grade 3 or 4 and difficult intubation. The search identified twelve high quality RCTs with 4,862 patients for inclusion. Among four studies reporting the primary outcome, there was no difference (RR=1.18; 95%CI=0.71 to 1.96; I2=0%; p=0.51). Only 3 studies were in patients at high risk of aspiration. There was significantly worse first attempt success (RR= 0.94; 95%CI= 0.89 to 0.99; I2=66%; p=0.02), time to intubation (WMD= 6.77seconds; 95%CI=4.40 to 9.14seconds; I2=97%) and laryngoscopy views (RR=1.69; 95%CI=1.41 to 2.02;I2=1%; p<0.00001) with cricoid pressure. Cricoid pressure failed to show any increase in protection from aspiration and may increase difficulty of intubation. Further studies in high-risk patients, such as intensive care patients, are required.


Simulating prosopagnosia through a lesion of lateral connections in a feed-forward neural network.
We show that particular features of prosopagnosic impairment can be simulated by a connectionist model trained with an unsupervised learning procedure. In particular we describe a Kohonen's neural network which is able to correctly recognize and categorize a series of digitized pictures of faces when learning is characterized by certain parameter values, but which shows a prosopagnosic behavior when lateral connections are lesioned. We discuss the relationship between this result and some neurophysiological hypotheses about prosopagnosia.


Epicardial adipose tissue and adrenal gland volume in patients with borderline personality disorder.
Borderline personality disorder (BPD) is associated with an elevated mortality risk that is partially attributed to suicide, but few studies examined other possible causes of premature death. The present study compared epicardial adipose tissue (EAT) volume as a known early predictor of premature cardiovascular morbidity, cardiovascular risk indices, and adrenal gland volume (AGV) as an indicator for chronic hypothalamus-pituitary-adrenal (HPA) axis activation in females with borderline personality disorder (BPD), major depressive disorder (MDD) and in healthy individuals. Twenty-eight patients with BPD comorbid with MDD (BPD/MDD), 22 MDD patients and 26 healthy females (CTRL) of comparable age were included. EAT and AGV were assessed by magnetic resonance tomography; 10-year cardiovascular risk and diabetes risk were determined by PROCAM and FINDRISK score; metabolic syndrome was defined following National Cholesterol Education Adult Treatment Panel III R (NCEP/ATP III) criteria. MADRS was used to assess depression severity. After adjustment for age, body mass index (BMI), and physical activity, EAT and AGV were significantly increased in BPD/MDD compared to MDD and CTRL. EAT and AGV displayed a positive correlation. Finally, diabetes risk in BPD/MDD was elevated compared to CTRL and MDD. The present study highlights the increased cardiometabolic risk of BPD patients. We identify EAT accumulation as an early predictor and potential mediator of cardiovascular disease in BPD that appears to be driven at least in part by HPA axis dysregulation. Therefore, interventions that reduce EAT volume (i.e. exercise and diet) should be considered in the clinical management of BPD.


Delayed and multiple hyperbaric oxygen treatments expand therapeutic window in rat focal cerebral ischemic model.
Although the brain-protective effect of single, early applications of hyperbaric oxygen (HBO) has been reported in acute ischemic stroke models, few studies have reported the long-term effect--especially after multiple HBO applications. This study employed delayed, multiple HBO treatments and evaluated cerebral infarction and neurological functional recovery for 4 weeks after transient focal ischemia. Adult male Sprague-Dawley rats were subjected to middle cerebral artery occlusion/reperfusion (MCAO/R) and were subsequently exposed to HBO (2.5 atmospheres absolute [ATA]) for 2 hours per day. HBO was administered at either 6 or 24 hours after MCAO/R and was repeated daily for 6 days. Rat behavior was scored to evaluate neurological deficits. The brains were removed for histological analysis of the infarct ratio at 1 and 4 weeks. Rats with HBO delayed for 6 or 24 hours following MCAO/R displayed a significant decrease of infarct ratio and amelioration of neurological deficits compared to the untreated group. This study suggests that delayed, but multiple, HBO treatments can improve neurological evaluation and reduce cerebral infarction.


Noninvasive ventilation in acute cardiogenic pulmonary edema: a meta-analysis of randomized controlled trials.
The evidence of individual studies in acute cardiogenic pulmonary edema (ACPE) supporting noninvasive ventilation (NIV) is still inconclusive, particularly regarding noninvasive positive pressure ventilation (NIPPV). We carried out a meta-analysis. We searched in the Embase, Medline, Cinahl, Dare, Coch, Central, and CNKI databases and congress abstracts for trials comparing continuous positive airway pressure (CPAP) or NIPPV with standard therapy (ST). To assess treatment effects, we carried out direct comparison using a random effects model and adjusted indirect comparison. At total of 34 studies (3,041 patients) were included. In direct comparisons, both CPAP and NIPPV reduced the risk of death (relative risk [RR] 0.64, 95% CI 0.44-0.93; RR 0.80, 95% CI 0.58-1.10; respectively) compared with ST, although only CPAP had a significant effect. There were no significant differences between NIPPV and CPAP. Pooled results of direct and adjusted indirect comparisons showed that compared with ST, both CPAP and NIPPV significantly reduced mortality (RR 0.63, 95% CI 0.44-0.89; RR 0.73, 95% CI 0.55-0.97; respectively). Our findings suggest that among ACPE patients, NIV delivered through either NIPPV or CPAP reduced mortality.


Regulatory mechanisms of long noncoding RNAs in vertebrate central nervous system development and function.
Long noncoding RNAs (lncRNAs) have emerged as an important class of molecules that regulate gene expression at epigenetic, transcriptional, and post-transcriptional levels through a wide array of mechanisms. This regulation is of particular importance in the central nervous system (CNS), where precise modulation of gene expression is required for proper neuronal and glial production, connection and function. There are relatively few functional studies that characterize lncRNA mechanisms, but possible functions can often be inferred based on existing examples and the lncRNA's relative genomic position. In this review, we will discuss mechanisms of lncRNAs as predicted by genomic contexts and the possible impact on CNS development, function, and disease pathogenesis. There is no doubt that investigation of the mechanistic role of lncRNAs will open a new and exciting direction in studying CNS development and function.


Successes and challenges of using whole exome sequencing to identify novel genes underlying an inherited predisposition for thoracic aortic aneurysms and acute aortic dissections.
Thoracic aortic aneurysms involving the aortic root and/or ascending aorta can lead to acute aortic dissections. Approximately 20% of patients with thoracic aortic aneurysms and dissections (TAAD) have a family history of the disease, referred to as familial TAAD (FTAAD) that can be inherited in an autosomal dominant manner with variable expression with respect to disease presentation, age of onset and associated features. Whole exome sequencing (WES) has been used to identify causative mutations in novel genes for TAAD. The strategy used to reduce the large number of rare variants identified using WES is to sequence distant relatives with TAAD and filter for heterozygous rare variants that are shared between the relatives, predicted to disrupt protein function and segregate with the TAAD phenotype in other family members. Putative genes are validated by identifying additional families with a causative mutation in the genes. This approach has successfully identified novel genes for FTAAD.


A review on the treatment of multiple myeloma with small molecular agents in the past five years.
Multiple myeloma is currently incurable, and the incidence rate is increasing year by year worldwide. Although in recent years the combined treatment plan based on proteasome inhibitors and immunomodulatory drugs has greatly improved the treatment effect of multiple myeloma, most patients still relapse and become resistant to current treatments. To solve this problem, scientists are committed to developing drugs with higher specificity, such as iberdomide, which is highly specific to ikaros and aiolos. This review aims to focus on the small molecular agents that are being researched/clinically used for the treatment of multiple myeloma, including the target mechanism, structure-activity relationship and application prospects of small molecular agents.


An immunochemical and immunohistochemical study of S100 protein in renal cell carcinoma.
The authors localized S100 protein in renal tubules and renal cell carcinoma by immunohistochemical study and quantitative analysis by enzyme immunoassay. The alpha subunit of S100 protein (S100-alpha) was localized in epithelial cells of proximal tubules, thin limbs of loops of Henle, collecting tubules, and a few of Bowman's capsules. The beta subunit (S100-beta) was immunostained in the distal tubules, thick and thin loops of Henle, collecting tubules, and a few proximal tubules. In renal cell carcinoma, S100-alpha was immunohistochemically demonstrated in 82% (41/50) of patients including sarcomatoid variants, whereas S100-beta was detected in 46% (23/50). Both the number of positively stained tumor cells and the staining intensity were greater in S100-alpha than in S100-beta. Concentrations of S100-alpha in the cortex were 80.3 +/- 22.5 ng/mg protein (n = 7), whereas those of renal cell carcinoma were 387 +/- 533 ng/mg protein (n = 19), i.e., about five times higher. Concentrations of S100-beta in both normal kidney (1.96 +/- 0.74 ng/mg protein) and renal cell carcinoma (2.05 +/- 2.16 ng/mg protein) were much lower than those of S100-alpha. The authors also localized S100-alpha and S100-beta in tissues of other renal tumors and tumors arising from other organs. S100a0 appears to be a useful immunohistochemical marker for renal cell carcinoma.


Comparison of three calibration procedures for free-field reference speech audiometry.
In standards IEC 60645-1 and ANSI S3.6, the free-field equivalent earphone output level method is assumed as the reference for speech audiometry. Three calibration procedures for this method were compared in this study. Speech audiometry was conducted with Dutch consonant-vowel-consonant words for the following conditions: 1. TDH39 earphones, 2. loudspeaker, and 3. free-field simulated with TDH39 earphones. The first calibration procedure was based on the empirically determined difference between the speech recognition threshold (SRT) with earphones and a loudspeaker. The second procedure was based on the theoretical free-field correction, derived from the known speech spectrum and the free-field to coupler difference. The third calibration procedure corresponded to the results of the free-field simulated speech material under earphones. The sample included 20 normal hearing subjects. The differences between the observed SRT in the free-field and earphone conditions and the free-field and simulated free-field conditions were 7.1 dB and 0.6 dB, respectively. The three calibration procedures for the free-field equivalent output method yielded approximately the same results, and therefore all appear to be useful for TDH39 earphones.


Synthesis of recurrent neural dynamics for monotone inclusion with application to Bayesian inference.
We propose a top-down approach to construct recurrent neural circuit dynamics for the mathematical problem of monotone inclusion (MoI). MoI in a general optimization framework that encompasses a wide range of contemporary problems, including Bayesian inference and Markov decision making. We show that in a recurrent neural circuit/network with Poisson neurons, each neuron's firing curve can be understood as a proximal operator of a local objective function, while the overall circuit dynamics constitutes an operator-splitting system of ordinary differential equations whose equilibrium point corresponds to the solution of the MoI problem. Our analysis thus establishes that neural circuits are a substrate for solving a broad class of computational tasks. In this regard, we provide an explicit synthesis procedure for building neural circuits for specific MoI problems and demonstrate it for the specific case of Bayesian inference and sparse neural coding.


[Microelectrophysiologic analysis of the representation of the vagal, splanchnic, sciatic nerves and sensomotor region of the cortex in the central nucleus of the amygdala].
In anesthetized cats, a high degree of reactivity (82.2%) of neurons was found to visceral nerves and visceral stimulation (single shocks and 2, 5, 10, 20 Hz) in the central nucleus of amygdala. The majority of the neurons revealed tonic changes of spontaneous activity. A high degree (76.6%) of afferent convergence from somato-visceral systems and cortical area was found.


Preparation of Male Meiotic Chromosomes for Fluorescence In Situ Hybridization and Immunodetection with Major Focus on Dogroses.
Meiosis is a very essential cell division resulting in the formation of four haploid gametes in plants. The preparation of meiotic chromosomes is a key step in plant meiotic research. Well-spread chromosomes, low background signal, and effective cell wall elimination give the best hybridization results. Dogroses (Rosa, section Caninae) are allopolyploids and frequently pentaploids (2n = 5x = 35) with asymmetrical meiosis. Their cytoplasm is enriched with organic compounds such as vitamins, tannins, phenols, essential oils, and many more. The cytoplasm is often a huge problem, avoiding successful cytogenetic experiments using fluorescence staining techniques. Here, we present a protocol with modifications for the preparation of male meiotic chromosomes suitable for fluorescence in situ hybridization (FISH) and immunolabeling with a major focus on dogroses.


Analysis of the 237Np-233Pa photon spectrum using the full response function method.
A study has been made of X- and gamma-ray emission from 237Np in equilibrium with 233Pa using the full response function method. This analysis process is characterised by photon spectrometry in which the entire spectrum is modelled in a pseudo-empirical way by means of elementary functions describing the total absorption and escape peaks, the Compton diffusion internal and external to the detector and the peaks resulting from detection of internal conversion electrons. This method has been applied to determine the L X-, K X- and gamma-rays emission probabilities in 237Np and 233Pa decay studies.


Auditory prediction errors and auditory white matter microstructure associated with psychotic-like experiences in healthy individuals.
Our sensory systems actively predict sensory information based on previously learnt patterns, which are continuously updated with information from the actual sensory input via prediction errors. Individuals with schizophrenia consistently show reduced auditory prediction errors as well as altered fractional anisotropy (indicative of white matter changes) in the arcuate fasciculus and the auditory interhemispheric pathway, both of which are auditory white matter pathways associated with prediction errors. However, it is not clear if healthy individuals with psychotic-like experiences exhibit similar deficits. Participants underwent electroencephalography (EEG) recordings while listening to a classical two-tone duration deviant oddball paradigm (n = 103) and a stochastic oddball paradigm (n = 89). A subset of participants (n = 89) also underwent diffusion-weighted magnetic resonance imaging (MRI). Fractional anisotropy (FA), was extracted from the arcuate fasciculi and the auditory interhemispheric pathway. While prediction errors evoked by the classical oddball paradigm failed to reveal significant effects, the stochastic oddball paradigm elicited significant clusters at the typical mismatch negativity time window. Furthermore, we observed that FA of the arcuate fasciculi and auditory interhemispheric pathway significantly improved predictive models of psychotic-like experiences in healthy individuals over and above predictions made by auditory prediction error responses alone. Specifically, we observed that decreasing FA in the auditory interhemispheric pathway and reducing ability to learn stochastic irregularities are associated with increasing CAPE + scores. To the extent that these associations have previously been reported in patients with schizophrenia, the findings from this study suggest that both, auditory prediction errors and white matter changes in the auditory interhemispheric pathway, may have the potential to be translated into early screening markers for psychosis.


Lattice Boltzmann Modeling of Drying of Porous Media Considering Contact Angle Hysteresis.
Drying of porous media is governed by a combination of evaporation and movement of the liquid phase within the porous structure. Contact angle hysteresis induced by surface roughness is shown to influence multi-phase flows, such as contact line motion of droplet, phase distribution during drainage and coffee ring formed after droplet drying in constant contact radius mode. However, the influence of contact angle hysteresis on liquid drying in porous media is still an unanswered question. Lattice Boltzmann model (LBM) is an advanced numerical approach increasingly used to study phase change problems including drying. In this paper, based on a geometric formulation scheme to prescribe contact angle, we implement a contact angle hysteresis model within the framework of a two-phase pseudopotential LBM. The capability and accuracy of prescribing and automatically measuring contact angles over a large range are tested and validated by simulating droplets sitting on flat and curved surfaces. Afterward, the proposed contact angle hysteresis model is validated by modeling droplet drying on flat and curved surfaces. Then, drying of two connected capillary tubes is studied, considering the influence of different contact angle hysteresis ranges on drying dynamics. Finally, the model is applied to study drying of a dual-porosity porous medium, where phase distribution and drying rate are compared with and without contact angle hysteresis. The proposed model is shown to be capable of dealing with different contact angle hysteresis ranges accurately and of capturing the physical mechanisms during drying in different porous media including flat and curved geometries. The online version contains supplementary material available at 10.1007/s11242-021-01644-9.


Docking, synthesis, in-vitro evaluation, and optimization of reaction conditions for direct radiolabeling of CGPRPPC with technetium-99m through the GAGG sequence.
With respect to the reported promising results of cyclic peptide CGPRPPC in early detection of thrombotic lesions, we developed a practical approach for technetium-99m labeling of this peptide using the Glycine-Alanine-Glycine-Glycine (GAGG) sequence as a chelating moiety. The peptide conjugated to GAGG was prepared using the solid-phase method. The optimization of radiolabeling conditions was performed on the basis of such variables as incubation time, reaction temperature, pH, and concentration of peptide and stannous chloride. Moreover, the stability and fibrin-binding affinity of the radiolabeled peptide were measured. The peptide-fibrin interactions were analyzed by docking studies using HEX and Auto dock 4.2. Softwares. The amounts of synthesized peptide and stannous chloride required for optimal radiolabeling through GAGG were 10 µmol/l and 5 µg, respectively. The best radiochemical purity% (>93%) was achieved at pH 7-8 within 15 min and a reaction temperature of 37°C. On the basis of in silico and in-vitro results, the GAGG-conjugated CGPRPPC peptide showed better binding affinity versus the HYNIC-conjugated one. We could radiolabel the fibrin-targeting peptide with high radiochemical purity% and stability during a short incubation period without a boiling step. Compared with the HYNIC-conjugated peptide, a higher binding affinity was found. Therefore, the GAGG chelating moiety possesses a considerable potentiality in Technetium 99m labeling of peptides while CGPRPPC maintains its binding properties to thrombotic lesions.


Clinical negligence claims following the COVID-19 pandemic.
John Tingle, Lecturer in Law, Birmingham Law School, University of Birmingham, discusses how the Courts may view clinical negligence claims brought by families who have lost loved ones during the pandemic.


Lack of enhanced cytotoxicity of cultured L1210 cells using folinic acid in combination with sequential methotrexate and fluorouracil.
Previous studies from this laboratory have demonstrated that treatment of cultured cells with sequential methotrexate (MTX) and fluorouracil (FUra) leads to synergistic cell killing in several murine and human neoplasms in vitro. In this study leucovorin (folinic acid, LCV) was added to the MTX/FUra combination with the intention of generating elevated levels of methylenetetrahydrofolate to promote the formation of a stable fluorodeoxyuridylate-thymidylate synthetase ternary complex, thereby augmenting the cytotoxicity of the MTX-FUra sequence. The addition of 10 or 100 microM LCV concurrently with or after 10 microM FUra following MTX (1 microM) pretreatment did not augment the inhibition of L1210 cell growth or the clonigenicity compared with MTX prior to FUra without LCV. The effects of LCV scheduling on the sequential MTX and FUra-induced inhibition of thymidylate synthesis were measured by examining the rate of [6-3H] dUrd incorporation into the acid-precipitable cell fraction and by direct quantitation of the thymidylate synthetase ternary complex. Combination of 100 microM LCV with 10 microM FUra after 1 microM MTX resulted in significantly more ternary complex formation than did 1 microM MTX before 10 microM FUra alone. The inhibitory effects of FUra on thymidylate synthetase in the presence of MTX, however, could not be augmented by LCV as determined by [6-3H] incorporation into acid-precipitable material, nor did the addition of LCV result in increased cytotoxicity. Factors other than the inhibition of DNA synthesis may be critical to the cytotoxicity of sequential MTX and FUra in L1210 cells.


Effect of salicylic acid elicitor on antioxidant potential and chemical composition of in vitro raised plants of Berberis asiatica Roxb. ex DC.
The present study for the first time investigated the effects of Salicylic acid (SA) (50, 100 and 200 mg/L) on in vitro growth and antioxidant capacity of Berberis asiatica. SA significantly enhanced in vitro shoot multiplication at 100 mg/L concentration. The result showed highest significant increase in antioxidant activity by 50 mg/L SA elicited plantlets in terms of DPPH (2,2-Di-phenyl-1-picryl-hydrazyl) free radical scavenging activity (IC50 32.42 ± 0.51), Ferric Reducing Antioxidant Activity (292.73 ± 2.09 mg AAE/g dw) and Metal Chelating Activity (IC50 13.18 ± 0.52). The Gas Chromatography-Mass Spectrometry (GC-MS) profiling revealed presence of 46, 38, 34 and 29 phytochemicals in control, SA1, SA2 and SA3 methanolic extracts respectively. Among the identified compounds, eight compounds, 4H-Pyran-4-one,2,3-dihydro-3,5-dihydroxy-6-methyl; 1,3,4,5-tetrahydroxy-cyclohexanecarboxylic acid; 3,7,11,15-Tetramethyl-2-hexadecen-1-ol (and Phytol); hexadecanoic acid, methyl ester; 9,12-Octadecadienoic acid (Z,Z)-,methyl ester; di-n-octyl phthalate; stigmast-5-en-3-ol,(3.beta.)- or ß- sitosterol; and squalene were previously known for antioxidant potential.


[New DVO guideline for osteoporosis management 2014 and its importance for trauma surgeons].
Osteoporosis-associated fractures represent a growing challenge in the treatment of orthopedic patients. In November 2014 a new revision of the guidelines on osteoporosis by the German Osteology Society (Dachverband Osteologie DVO) was adopted, in which additional risk factors for fractures and further treatment options have been included. On the one hand the existing model used to diagnose osteoporosis and estimate a high fracture risk as a guidance for the use of specific anti-osteoporotic therapy in patients without a fragility fracture was maintained and further refined. On the other hand the guideline includes the option to initiate a specific osteoporosis therapy without a prior bone densitometry in patients with typical radiographs of a proximal femur fracture and higher grade vertebral fractures, suspicious for osteoporosis, depending on the overall clinical context. This may reduce the treatment gap of osteoporosis in Germany. In this paper the changes in the DVO guidelines 2014 on osteoporosis are summarized, focusing on the most important changes with practical relevance for orthopedic surgeons.


Brain-specific gene expression.
The brain of an adult rat expresses approximately 30,000 different brain-specific mRNAs. To investigate their encoded proteins, we have selected cDNA clones corresponding to mRNAs expressed exclusively in rat brain, determined their nucleotide sequences and generated antisera against synthetic peptides mimicking short regions of the deduced protein sequences. The clone plB236 encodes a protein that defines a widely distributed neuronal system and may be the precursor for a family of novel neuropeptides. A second clone, plB208, encodes rat brain proteolipid protein, the major protein component of central nervous system myelin. These studies have also identified an 82 nucleotide genetic element called an ID (identifier) sequence that may be involved in the regulation of transcription of brain-specific genes.


Adult brain aging investigated using BMC-mcDESPOT-based myelin water fraction imaging.
The relationship between regional brain myelination and aging has been the subject of intense study, with magnetic resonance imaging perhaps the most effective modality for elucidating this. However, most of these studies have used nonspecific methods to probe myelin content, including diffusion tensor imaging, magnetization transfer ratio, and relaxation times. In the present study, we used the BMC-mcDESPOT analysis, a direct and specific method for imaging of myelin water fraction (MWF), a surrogate of myelin content. We investigated age-related differences in MWF in several brain regions in a large cohort of cognitively unimpaired participants, spanning a wide age range. Our results indicate a quadratic, inverted U-shape, relationship between MWF and age in all brain regions investigated, suggesting that myelination continues until middle age followed by decreases at older ages. We also observed that these age-related differences vary across different brain regions, as expected. Our results provide evidence for nonlinear associations between age and myelin in a large sample of well-characterized adults, using a direct myelin content imaging method.


Meta-analysis confirmed genetic susceptibility conferred by multiple risk variants from CTLA4 and SERPINA1 in granulomatosis with polyangiitis.
Granulomatosis with polyangiitis (GPA) is a rare systemic autoimmune disease. Smaller sample size and complex nature of the disease pathogenesis has made it challenging to perform well-powered genetic investigations. We performed a systematic review based meta-analysis in GPA to investigate the genetic susceptibility conferred by non-human leukocyte antigen (non-HLA) candidate genes. A systematic review was performed using web-based literature search and eligible studies were included following inclusion-exclusion criteria. Studies were evaluated for their quality of evidence and study outcome was assessed using the Newcastle-Ottawa Scale and Grades of Research, Assessment, Development and Evaluation tools. Reviewer's agreement was accessed through Cohen's κ value. Meta-analyses were performed using RevMan 5 tool. Meta-odds ratio (meta-OR) and Z test P value were evaluated to estimate the genetic susceptibility for each of the variants. Eighteen studies were found eligible and 7 genetic variants from only 4 genes, namely CTLA4, PRTN3, SERPINA1 and PTPN22 could be studied for meta-analysis. rs231775-G (49-G) (Meta-OR = 1.42 [1.14-1.76]; P = .001) of CTLA4 and rs7151526-A (Meta-OR = 2.70 [1.51-4.85]; P = .0008) of SERPINA1 were confirmed to be predisposing alleles, and rs5742909-C (318-C) (Meta-OR = 0.65 [0.44-0.97]; P =.03) of CTLA4 was found to be protective for GPA. In concordance with the genetic association of rs7151526-A, serological marker for the same variant "Z" allele of SERPINA1 was found to be predisposing (Meta-OR = 12.60 [5.01-31.68]; P < .00001) for GPA. Genetic variants confirmed in this study play critical roles in T-cell mediated immune function and could be significantly implicated in GPA. Molecular pathology studies are warranted to confirm their role. These markers could be used for efficient patient classification and disease management.


An investigation of the metabolism of N-nitroso-N-methylaniline by phenobarbital- and pyrazole-induced Sprague-Dawley rat liver and esophagus derived S-9.
The metabolism and mutagenicity of the esophageal carcinogen N-nitroso-N-methylaniline (NMA) was studied using hepatic and esophageal 9000 X g supernatant (S-9) preparations from Sprague-Dawley rats induced with pyrazole and phenobarbital. Only pyrazole-induced hepatic S-9 was able to dose-dependently activate NMA to a mutagen in the Ames assay and specifically in Salmonella typhimurium TA1537. NMA in the presence of phenobarbital-induced S-9 gave a very weak non-dose dependent mutagenic response. Metabolism of NMA by the two induced hepatic and esophageal S-9 fractions yielded aniline and N-methylaniline (MA). Phenobarbital-induced S-9 from both tissues also afforded phenol, while none was found with the pyrazole-induced preparations. Phenol formation presumably arose from the direct oxidative demethylation of NMA via a benzenediazonium ion (BDI) intermediate. The results indicate that an important metabolic pathway for NMA, with both inducing agents, entails an initial denitrosation to yield MA, which in turn rapidly undergoes oxidative demethylation to aniline. The conversion of NMA to phenol also suggests that direct demethylation of NMA in the phenobarbital-induced system is an important metabolic pathway.


Calcaneal osteotomy due to insertional calcaneal tendinopathy: preoperative planning.
Dorsal closing wedge calcaneal osteotomy (DCWCO) is indicated in patients with insertional tendinopathy of the calcaneal (Achilles) tendon. The Chauveaus-Liet's (CL) angle is represented by the difference between the angle of verticalization (α) and morphological angle (β) of the calcaneus (CL angle = α - β). The purpose of the study was to assess whether the DCWCO affects the Chauveaus-Liet's angle. The study included 12 patients indicated to DCWCO. Three directions of close wedge osteotomy were designed for each patient-horizontal, vertical and in the middle type of osteotomy and a virtual osteotomy was created in each of them in the ABAQUS system in cooperation with Czech Technical University. The most used directions of osteotomy according to the available literature were used. We evaluated α and β angles before and after osteotomy, changes of the length plantar aponeurosis and the elevation of distal insertional point of the calcaneal tendon. The changes of grades, median and standard deviation were observed. The change of the alfa angle was dependent on the direction of the osteotomy and the change of the beta angle was affected by the size of the osteotomy. The greatest elevation of the distal insertional point of the calcaneal tendon occurred in the horizontal type of the osteotomy. Our study shows that the more we want to reduce the tension in the calcaneal tendon, the more we have to perform an osteotomy horizontally. This study could serve as a preoperative guide for osteotomy planning.


Crosstalk between hepatocyte growth factor and integrin signaling pathways.
Most types of normal cells require integrin-mediated attachment to extracellular matrix to be able to respond to growth factor stimulation for proliferation and survival. Therefore, a consensus that integrins are close collaborators with growth factors in signal transduction has gradually emerged. Some integrins and growth factor receptors appear to be normally in relatively close proximity, which can be induced to form complexes upon cell adhesion or growth factor stimulation. Moreover, since integrins and growth factor receptors share many common elements in their signaling pathways, it is clear tzhat there are many opportunities for integrin signals to modulate growth factor signals and vice versa. Increasing evidence indicates that integrins can crosstalk with receptor tyrosine kinases in a cell- and integrin-type-dependent manner through a variety of specific mechanisms. This review is intended specifically for summarizing recent progress uncovering how the hepatocyte growth factor receptor c-Met coordinates with integrins to transmit signals.


Changes in the Expression of Transcription Factor Oct4 during Postnatal Development of Adrenal Medulla.
We analyzed the expression of the transcription factor Oct4 in the chromaffin cells during the postnatal development of the adrenal glands in rats. Single Oct4+-chromaffin cells with nuclear localization of the protein were found in the medulla, and their number increased in parallel with a decrease in the proliferative activity of cells. In postnatal development, 100% of chromaffin cells demonstrated high expression of tyrosine hydroxylase, which attested to their differentiation and functional activity. It was found that all Oct4+ cells were differentiated chromaffin cells. An increase in the pool of Oct4-expressing cells after the completion of organ growth suggests the formation of a potential source for the physiological and reparative regeneration of the adrenal medulla.


Self-reported skin morbidity and mental health. A population survey among adults in a Norwegian city.
Several studies among dermatological patients have shown a link between various chronic dermatological diseases and mental morbidity. To explore the association between self-reported skin morbidity and psychosocial factors in the general population. This population-based cross-sectional study is part of the Oslo Health Study conducted during 2000-2001. All individuals in Oslo County, Norway, born in 1924/25, 1940/41, 1955, 1960 and 1970 received a postal questionnaire, which 18,770 men and women answered. The questionnaire provided information on sociodemographic factors and self-reported health and psychosocial factors. Dichotomous variables for 10 self-reported skin complaints were used. These were previously validated and refer to the most common chronic skin diseases. Mental distress was measured with a validated 10-item instrument, the Hopkins Symptom Check List-10; social support with the number of confidants; and negative life events with a 12-item validated instrument. The odds ratio (OR) for mental distress was 1.70 [95% confidence interval (CI) 1.21-2.38] for having itch, 1.64 (95% CI 1.15-2.34) for pimples and 1.72 (95% CI 1.06-2.80) for face rash in an adjusted model. In an adjusted model the OR for skin disease was 1.60 (95% CI 1.39-1.84) when the individual had experienced more than two negative life events; and 2.52 (95% CI 2.12-3.00) for mental distress. Skin morbidity increased for both genders, with poor social support network. There was a significant interaction between social support network and negative life events in the logistic regression model for skin disease when adjusted for sociodemographic factors. The study quantifies the association between dermatological problems and psychosocial factors at a population level. It underlines the need to focus on these issues in research and needs assessment in dermatology.


Glycosylated analogs of formaecin I and drosocin exhibit differential pattern of antibacterial activity.
The synthetic glycopeptides are interesting model systems to study the effect of O-glycosylation in modulating their function and structure. A series of glycosylated analogs of two antibacterial peptides, formaecin I and drosocin, were synthesized by varying the nature of sugar and its linkage with bioactive peptides to understand the influence of structure variation of glycosylation on their antibacterial activities. Higher antibacterial activities of all glycopeptides compared to their respective non-glycosylated counterparts emphasize in part the importance of sugar moieties in functional implications of these peptides. The consequences of the unique differences among the analogs were apparent on their antibacterial activities but not evident structurally by circular dichroism studies. We have shown that differently glycosylated peptides exhibit differential effect among each other when tested against several Gram-negative bacterial strains. The change of monosaccharide moiety and/or its anomeric configuration in formaecin I and drosocin resulted into decrease in the antibacterial activity in comparison to that of the native glycopeptide, but the extent of decrease in antibacterial activity of glycosylated drosocin analogs was less. Probably, the variation in peptide conformation arising due to topological dissimilarities among different sugars in the same peptide resulting in possible modulation in binding properties appears to be responsible for differences in their antibacterial activities. Indeed, these effects of glycosylation are found to be sequence-specific and depend in the milieu of amino acid residues. Interestingly, none of the carbohydrate variants affected the basic property of these peptides, which is non-hemolytic and non-toxicity to eukaryotic cells.


Ascorbic acid-dependent collagen formation in penaeid shrimp.
1. This study tested the hypothesis that black death, the ascorbic acid (AsA) related disease of penaeid shrimp, is related to collagen underhydroxylation. 2. Collagen measured as hydroxyproline (HYP) in healthy Penaeus californiensis (Holmes) and P. stylirostris (Stimpson) of a wide range of masses were determined. The results revealed a logarithmic relationship between total body collagen HYP and body weight fitting the equation y = 90x1.18 where y = total collagenous HYP (microgram) and x = body weight (g). 3. Shrimp tissues most subject to mechanical trauma (subcutis, hindgut and gills) had the highest collagenous HYP levels and were most consistently and severely affected by an ascorbic acid (AsA) deficiency disease. 4. Prolyl hydroxylase (PH) activity was demonstrated in tissues of P. californiensis and P. stylirostris by hydroxylation of [3,4-3H]proline. 5. AsA was required for shrimp PH activity using a chicken embryo substrate. 6. Nutritional trials revealed that dietary AsA was required for proline hydroxylation in collagen formation in P. californiensis.


How complexity science can inform a reflective process for improvement in primary care practices.
Quality improvement processes have sometimes met with limited success in small, independent primary care settings. The theoretical framework for these processes uses an implied understanding of organizations as predictable with potentially controllable components. However, most organizations are not accurately described using this framework. Complexity science provides a better fit for understanding small primary care practices. The Multimethod Assessment Process (MAP)/Reflective Adaptive Process (RAP) is informed by complexity science. This process was developed in a series of studies designed to understand and improve primary care practice. A case example illustrates the application and impact of the MAP/RAP process. Guiding principles for a reflective change process include the following: an understanding of practices' vision and mission is useful in guiding change, learning and reflection helps organizations adapt to and plan change, tension and discomfort are essential and normal during change, and diverse perspectives foster adaptability and new insights for positive change. A reflective change process that treats organizations as complex adaptive systems may help practices make sustainable improvements.


Concordance between current job and usual job in occupational and industry groupings: assessment of the 2010 national health interview survey.
To determine whether current job is a reasonable surrogate for usual job. Data from the 2010 National Health Interview Survey were utilized to determine concordance between current and usual jobs for workers employed within the past year. Concordance was quantitated by kappa values for both simple and detailed industry and occupational groups. Good agreement is considered to be present when kappa values exceed 60. Overall kappa values ± standard errors were 74.5 ± 0.5 for simple industry, 72.4 ± 0.5 for detailed industry, 76.3 ± 0.4 for simple occupation, 73.7 ± 0.5 for detailed occupation, and 80.4 ± 0.6 for very broad occupational class. Sixty-five of 73 detailed industry groups and 78 of 81 detailed occupation groups evaluated had good agreement between current and usual jobs. Current job can often serve as a reliable surrogate for usual job in epidemiologic studies.


The complex interplay of physician, patient, and spouse in preoperative counseling for radical prostatectomy: a comparative mixed-method analysis of 30 videotaped consultations.
Spouses of cancer patients play a crucial role in deciding on therapeutic choices. The aim of our study was to assess their role in counseling for radical prostatectomy. We analyzed 30 videotaped preoperative consultations prior to radical prostatectomy. Thereof, 14 included the patients' female partner and 16 took place without partner attendance. We performed quantitative and qualitative conversation analysis to compare both settings. Mean age of patients was 61 (47-73) years; 13% (4/30) did not have a partner. Duration of preoperative consultations was 20 (10-32) min. Physicians spoke most of the time (93%, range 71-99%), followed by patients (7%, range 1-20%) and spouses (2%, range 0-8%). Patients whose spouse was present at the consultation tended to have a more averted posture (50% vs. 25%, p = 0.04) and tended to speak less often (5% vs. 8%, p = 0.02). In 4 of 14 (29%) consultations, the spouses tended to be more dominant, speaking more frequently. Qualitative analysis showed several examples of emotional support and helpful contributions by spouses. Difference of opinion occurred when pros and cons of a nerve-sparing approach were discussed. The spouses' impact appeared to influence the final decision of men contemplating a nerve-sparing approach in 1 of 14 conversations. Spouses appear to play a complex and sometimes ambivalent role in counseling for radical prostatectomy. Especially when discussing a nerve-sparing approach, urologist should focus on the patients' true needs while interacting with both partners. Personalized decision aids might help to identify possible conflicts in advance.Copyright © 2015 John Wiley & Sons, Ltd.


Effect of thyrotropin-releasing hormone on immune functions of peripheral blood mononuclear cells.
The tripeptide thyrotropin-releasing hormone (TRH) works as a hypothalamic hormone, but is found also outside the brain in intrinsic nerve fibers of the gastrointestinal tract. There is evidence that TRH modulates the activity of immunocompetent cells, although there are only very few data on TRH-mediated immune effector functions. Since we could recently show that TRH inhibits monocyte activities we were also interested in other possible TRH modulated immune functions. Peripheral blood mononuclear cells (PBMC) from ten healthy subjects were cultured for 7 days and pulsed with 0.125 and 0.250 microgram/ml Pokeweed mitogen (PWM). 10(-12) to 10(-6) M TRH was added simultaneously with PWM. Lymphocyte proliferation [(3H]thymidine incorporation), interferon-gamma (IFN-gamma) activity (RIA) and immunoglobulin activities (IgG, IgM, IgA; ELISA) were determined in the supernatants. We could demonstrate a TRH-dependent decrease in PWM-pulsed IgG activity with significant (alpha = 0.05) values at 10(-8) and 10(-10) M (-29 +/- 6%/-16 +/- 3% for PWM 0.125 microgram/ml and -17 +/- 9%/-11 +/- 9% for PWM 0.250 microgram/ml). This inhibitory effect could be abolished by an anti-TRH antiserum. There was no TRH effect on IgM and IgA activities, IFN-gamma activity and lymphocyte proliferation compared with the PWM stimulated values alone. The described TRH effect on the polyclonal IgG response by PBMC gives further evidence for a functional link between the immune system and the endocrine system, although its underlying mechanism is not yet clear.


Evisceration with implantation of an intrascleral silicone prosthesis in dogs: A retrospective study of 107 cases (2010-2019).
To retrospectively analyze the preoperative clinical characteristics and surgical outcomes of evisceration with implantation of an intrascleral silicone prosthesis (EIISP) procedures in dogs and evaluate whether brachycephalic dogs are more prone than non-brachycephalic dogs to develop postoperative complications after EIISP. Ninety-One dogs (19 of which were brachycephalic) were included. Medical records from 2010 to 2019 were reviewed. Signalment, reason for EIISP, postoperative complications, follow-up time, and postoperative eye appearance were analyzed. The most frequently represented breeds were French Bulldog [11/91 (12%) dogs], Jack Russell Terrier [6/91 (7%)], and Shih Tzu [6/91 (7%)]. Brachycephalic dogs were statistically younger than non-brachycephalic dogs at the time of EIISP (p = 1.61 × 10-5 ). Uncontrolled glaucoma was the most common reason for EIISP in both groups. Short-term complications (from D0 to D15) seen in 7/91 (8%) dogs included epithelial corneal ulcers (n = 3), keratoconjunctivitis sicca (KCS) (n = 2), and prosthesis extrusion (n = 2). Long-term complications seen in 26/91 (29%) dogs included KCS (n = 11), epithelial corneal ulcers (n = 7), stromal ulcers (n = 3), entropion (n = 4), and prosthesis extrusion (n = 1). Extrusion of the prosthesis occurred twice in eyes that had undergone diode laser transscleral cyclophotocoagulation. The risk of postoperative complications was not significantly different between brachycephalic and non-brachycephalic dogs (p = .3). Brachycephalic status in dogs does not appear to influence the risk of complications from EIISP. Nevertheless, the present study is a reminder that EIISP focuses on esthetics, and considering the possible complications associated with it, it does not provide a benefit to the patient compared to enucleation.


Nucleomorphometric analysis of canine cutaneous mast cell tumours.
Thirty-five canine cutaneous mast cell tumours (CCMCTs) were analysed by computerized nuclear morphometry. In each case, the nuclei of at least 100 neoplastic cells were measured, and the mean nuclear area (MNA), mean nuclear perimeter (MNP) and mean nuclear form factor (FF) were calculated. Significant differences in respect of MNA and MNP occurred between tumours of grades I and III and between those of grades II and III (P<0.01) but not between tumours of grades I and II (P>0.01). No significant differences in respect of FF were observed between tumours of different grades. The results obtained indicate that nuclear morphometric analysis may assist in the grading of CMCTs.


GH and Pituitary Hormone Alterations After Traumatic Brain Injury.
Traumatic brain injury (TBI) is a crucially important public health problem around the world, which gives rise to increased mortality and is the leading cause of physical and psychological disability in young adults, in particular. Pituitary dysfunction due to TBI was first described 95 years ago. However, until recently, only a few papers have been published in the literature and for this reason, TBI-induced hypopituitarism has been neglected for a long time. Recent studies have revealed that TBI is one of the leading causes of hypopituitarism. TBI which causes hypopituitarism may be characterized by a single head injury such as from a traffic accident or by chronic repetitive head trauma as seen in combative sports including boxing, kickboxing, and football. Vascular damage, hypoxic insult, direct trauma, genetic predisposition, autoimmunity, and neuroinflammatory changes may have a role in the development of hypopituitarism after TBI. Because of the exceptional structure of the hypothalamo-pituitary vasculature and the special anatomic location of anterior pituitary cells, GH is the most commonly lost hormone after TBI, and the frequency of isolated GHD is considerably high. TBI-induced pituitary dysfunction remains undiagnosed and therefore untreated in most patients because of the nonspecific and subtle clinical manifestations of hypopituitarism. Treatment of TBI-induced hypopituitarism depends on the deficient anterior pituitary hormones. GH replacement therapy has some beneficial effects on metabolic parameters and neurocognitive dysfunction. Patients with TBI without neuroendocrine changes and those with TBI-induced hypopituitarism share the same clinical manifestations, such as attention deficits, impulsion impairment, depression, sleep abnormalities, and cognitive disorders. For this reason, TBI-induced hypopituitarism may be neglected in TBI victims and it would be expected that underlying hypopituitarism would aggravate the clinical picture of TBI itself. Therefore, the diagnosis and treatment of unrecognized hypopituitarism due to TBI are very important not only to decrease morbidity and mortality due to hypopituitarism but also to alleviate the chronic sequelae caused by TBI.


Thrombospondin-4 regulates fibrosis and remodeling of the myocardium in response to pressure overload.
Thrombospondin-4 (TSP-4) expression increases dramatically in hypertrophic and failing hearts in rodent models and in humans. The aim of this study was to address the function of TSP-4 in the heart. TSP-4-knockout (Thbs4(-/-)) and wild-type (WT) mice were subjected to transverse aortic constriction (TAC) to increase left ventricle load. After 2 wk, Thbs4(-/-) mice had a significantly higher heart weight/body weight ratio than WT mice. The additional increase in the heart weight in TAC Thbs4(-/-) mice was due to increased deposition of extracellular matrix (ECM). The levels of interstitial collagens were higher in the knockout mice, but the size of cardiomyocytes and apoptosis in the myocardium was unaffected by TSP-4 deficiency, suggesting that increased reactive fibrosis was the primary cause of the higher heart weight. The increased ECM deposition in Thbs4(-/-) mice was accompanied by changes in functional parameters of the heart and decreased vessel density. The expression of inflammatory and fibrotic genes known to be influential in myocardial remodeling changed as a result of TSP-4 deficiency in vivo and as a result of incubation of cells with recombinant TSP-4 in vitro. Thus, TSP-4 is involved in regulating the adaptive responses of the heart to pressure overload, suggesting its important role in myocardial remodeling. Our study showed a direct influence of TSP-4 on heart function and to identify the mechanism of its effects on heart remodeling.


Genome-wide analyses of early-onset acute myocardial infarction identify 29 novel loci by whole genome sequencing.
Early-onset acute myocardial infarction (AMI) may have a higher genetic predisposition than late-onset AMI. The present study aimed to identify and characterize germline variants that affect early-onset AMI using whole-genome sequencing (WGS). We performed a genome-wide association study based on the WGS of 1239 Koreans, including 596 early-onset AMI patients and 643 healthy individuals. Patients with AMI who underwent percutaneous coronary intervention (PCI) caused by atherothrombotic occlusive lesions were included in the study. A total of 29 novel loci were found to be associated with early-onset AMI. These loci are involved in thrombosis, fibrinolysis, inflammation, and lipid metabolism. One of the associated single nucleotide variants (SNVs), rs1614576, located upstream of PRKCB, is known to be associated with thrombus formation. Additionally, the results revealed a novel locus, rs78631167, located upstream of PLAUR which plays a critical role in regulating plasminogen activation and is related to fibrinolysis. The association between early-onset AMI and rs9357455, which is located upstream of PHACTR1 and regulates inflammation in AMI, was found. Moreover, we identified a lipid metabolism related genetic risk locus, rs5072, in the APOA1-AS gene. This study provides new evidence supporting the genetic association between early-onset AMI and thrombosis and fibrinolysis, as well as inflammation and lipid metabolism, by analyzing the whole-genome of 596 patients with early-onset AMI who have been treated with PCI. Our findings highlight potential genetic markers for the prediction and management of AMI, as well as for understanding the etiology of AMI.


Long-term health-related quality of life in living liver donors: A south Asian experience.
The aim of this study was to evaluate long-term health-related quality of life (HRQOL), changes in lifestyle, and complications in living liver donors at a single transplant center from southern India. A total of 64 consecutive living liver donors from 2008 to 2011 were evaluated; 46 of 64 donors completed the short form 36 (SF-36) via telephonic interviews or clinic consultations. Mean follow-up was 48 months (range: 37-84 months). There was no mortality in the donors evaluated. Overall morbidity was 23%, which included wound infections (4.3%), incisional hernia (2.1%), biliary leak (4.3%), and nonspecific complaints regarding the incision site (15.2%). All 46 donors who completed the SF-36 had no change in career path or predonation lifestyle. A total of 40 of 46 (87%) donors had no limitations, decrements, or disability in any domain, while six of 46 (13%) had these in some domains of which general health (GH) was most severely affected. Living donor hepatectomy is safe with acceptable morbidity and excellent long-term HRQOL with no change in career path or significant alteration of lifestyle for donors.


Effect of amiloride on bulk flow and iontophoretic taste stimuli in the hamster.
This investigation demonstrates the effect of amiloride on various taste responses in the hamster, and tests the hypothesis that its action on iontophoretic application of taste stimuli parallels its action on bulk flow delivery. Amiloride has not previously been tested in the hamster nor has its effect on iontophoretic stimuli (so-called 'electric taste'), which is thought to behave similarly to bulk flow stimuli, been examined. Amiloride treatment (4 min of 0.0001 M) of the hamster's tongue effectively inhibited chorda tympani responses to NaCl and LiCl solutions. Bulk flow (0.1 M) and iontophoretic (+ 7 microA through 0.001 M) presentations of NaCl and LiCl, which had unequal response magnitudes pre-treatment, were inhibited to the same residual response magnitude post-treatment. Recovery then proceeded along two distinct curves asymptotically returning to pre-treatment response levels. These curves could be adequately described by a simple exponential relationship. KCl responses were unaffected when presented via bulk flow techniques but significantly reduced when presented iontophoretically. HCl responses via either method were only slightly diminished. No decrement in response level was observed for the sweet stimuli sucrose (0.5 M) or saccharin (-9 microA through 0.001 M Na-saccharin) nor for potassium picrate, a bitter stimulus, (0.01 M or -10 microA through 0.001 M). Amiloride treatment of the hamster tongue was as specific in its action for sodium and lithium as reported in other species, and with the exception of KCl the action of amiloride on iontophoretic stimulation paralleled its action on bulk flow stimulation.


Generation of Recombinant Capripoxvirus Vectors for Vaccines and Gene Knockout Function Studies.
The ability to manipulate capripoxvirus through gene knockouts and gene insertions has become an increasingly valuable research tool in elucidating the function of individual genes of capripoxvirus, as well as in the development of capripoxvirus-based recombinant vaccines. The homologous recombination technique is used to generate capripoxvirus knockout viruses (KO), and is based on the targeting a particular viral gene of interest. This technique can also be used to insert a gene of interest. A protocol for the generation of a viral gene knockout is described. This technique involves the use of a plasmid which encodes the flanking sequences of the regions where the homologous recombination will occur, and will result in the insertion of an EGFP reporter gene for visualization of recombinant virus, as well as the E. coli gpt gene as a positive selection marker. If an additional gene is to be incorporated, this can be achieved by inserting a gene of interest for expression under a poxvirus promoter into the plasmid between the flanking regions for insertion. This chapter describes a protocol for generating such recombinant capripoxviruses.


Enhanced culture assay for detection and quantitation of latently infected, resting CD4+ T-cells carrying replication-competent virus in HIV-1-infected individuals.
Highly active antiretroviral therapy can decrease plasma HIV-1 levels to below the limit of detection. However, HIV-1 persists in latently infected resting-memory CD4+ T-cells carrying an integrated copy of the viral genome. The pool of latently infected cells is extremely stable and represents a major barrier to HIV-1 eradication. Identification and characterization of this reservoir required the development of methods for purifying resting CD4+ T-cells from HIV-1-infected individuals, activating the cells to induce virus production, and detecting and quantitating cells capable of releasing infectious virus. The development of an enhanced viral culture assay to quantitate the number of latently infected cells carrying replication competent virus is described here.


The intrauterine contraceptive device and acute salpingitis: a multifactor analysis.
From an indigent population hospitalized with acute salpingitis, 163 patients were compared with 222 control patients from the minor trauma section of the emergency room. Four factors were significantly different between cases and controls: race, type of contraception, number of sex partners, and previous history of salpingitis. A discriminate analysis based on a linear logistic equation demonstrated that each risk factor was independent rather than a reflection of another risk factor. Thus factors associated with race, IUD use, multiple sexual partners, and previous salpingitis increase the risk of salpingitis in the population studied.


Weight gain patterns during twin gestation.
Few data are available about desirable weight gain during twin gestation. The present study addresses this issue through retrospective evaluation of 217 women pregnant with twins who delivered at Swedish Hospital Medical Center, Seattle, between 1982 and 1986. The purpose of the study was to identify an optimum total weight gain and an optimum weight gain pattern for normal twin gestation. A mean prenatal weight gain of 20 kg (44 lb) was associated with "optimum" outcome; optimum outcome was defined as a pregnancy lasting at least 37 weeks with the delivery of two living infants weighing greater than or equal to 2,500 gm each with 5-minute Apgar scores greater than or equal to 7. A weight gain curve for twin gestation was generated and compared with the standard curve for singleton pregnancy. While the weight gain curve during twin gestation was similar to the standard curve during the first half of pregnancy, increased weight gain began in the second trimester and continued to term. The weight gain pattern of twin gestations with less-than-optimum outcome paralleled that of the optimum outcome group until about 30 weeks' gestation, at which time weight gain slowed in women in the former group, whose mean total weight gain was 16.8 kg (37 lb).