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1.
Article in English | MEDLINE | ID: mdl-37269524

ABSTRACT

Poria cocos has been dried in an indirect solar drying system composed of a roughened solar air heater (RSAH), a shell and tube storage unit assisted with flat micro heat pipes fins, and a drying chamber. The main novelty in this study is using FMHPs as fins in shell and tube storage unit with paraffin wax and lack of investigations on Poria cocos solar drying as medicinal material used in Chinese medicine. First and second laws of thermodynamics are used to assess the performance of the system and the results indicated that the RSAH average thermal ([Formula: see text]) and exergy efficiency ([Formula: see text]) were 73.9% and 5.1%, respectively, with averaged incident solar radiation of 671 W/m2 under airflow rate of 0.0381 m3/s. Furthermore, the storing system showed 37.6% as averaged overall [Formula: see text] and 17.2% as averaged overall [Formula: see text], as well as, discharging prolonged to 4 h with effective drying temperature. The overall [Formula: see text] of the dryer was 27.6% with specific energy consumption (SEC) of 8.629 kWh/kg moisture. The payback period of the system is 1.7 years.

2.
Eat Weight Disord ; 27(7): 2821-2834, 2022 Oct.
Article in English | MEDLINE | ID: mdl-35790669

ABSTRACT

PURPOSE: Sleep impairment is reported to be a consequence of overweight and obesity. However, the weight-sleep relationship can alternately be explained by demographics (e.g. age) and covariates (i.e. mood/affect and behaviour in overweight/obese people; e.g. night-eating). Thus, we examined the weight-sleep quality relationship after controlling for the effects of affect and common behaviour (i.e. night-eating, insufficient exercise, alcohol and electronic device use). METHODS: Online questionnaires asked 161 overweight, obese or normal-weight participants about their sleep quality, night-eating, physical activity, alcohol use, electronic device use and anxiety and depression at T0 (baseline) and T1 (3 months later). Height and weight and waist and hip circumference were objectively measured at T0 and T1, and physical activity was assessed over 24 h (using actigraphy) at T0 and T1. Hierarchical multiple regression analyses evaluated whether the weight measures (i.e. body-mass-index [BMI], waist-to-hip ratio [WHR] and obesity category [overweight/obese vs. normal-weight]) predicted sleep quality and its components at T0 and T1, after controlling demographics (at step 1) and covariates (affective distress and behaviour) at step 2, and entering weight measures at step 3; maximum 8 variables in the analyses. RESULTS: High BMI predicted several aspects of sleep quality after taking into account co-existing behaviour, affect and demographics: sleep disturbances at T0 and lower sleep efficiency at T1. WHR and obesity category did not predict any aspects of sleep quality. Several co-existing behaviour were related to or predicted sleep quality score and aspects of sleep quality including night-eating, alcohol use and electronic device use and affective symptoms (i.e. anxiety, depression). CONCLUSION: Results suggest that a person's weight may impact on their sleep quality above and beyond the effects of their co-existing behaviour and affect, although their co-existing behaviour and affect may also adversely impact on sleep quality. LEVEL OF EVIDENCE: Level III, evidence obtained from well-designed cohort.


Subject(s)
Overweight , Sleep Quality , Body Mass Index , Humans , Obesity/complications , Overweight/complications , Overweight/psychology , Sleep , Waist-Hip Ratio
3.
Br Poult Sci ; 63(6): 864-870, 2022 Dec.
Article in English | MEDLINE | ID: mdl-35791891

ABSTRACT

1. This paper details the establishment of a diagnostic system based on the real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the rapid, simple and sensitive detection of genotype VII of Newcastle disease virus (NDV) directly from field samples. One specific set of six primers were designed which targeted the fusion protein gene of G-VII viruses. The target gene can be amplified and the results recorded within 40 min.2. The merit of this technique was the feasibility of reading results either by examining turbidity by the naked eye or via the amplification curve generated by real-time PCR. This study tested the sensitivity and specificity of this system against NDV-G-VII and other avian viruses. The real-time RT-LAMP has been found to be more sensitive than real-time RT-PCR. Moreover, 24 out of 35 suspected field samples were positive for genotype VII by real-time RT-LAMP within 30 min in comparison to the real-time RT-PCR for detection of universal NDV.3. Accordingly, real-time RT-LAMP revealed higher sensitivity than real-time RT-PCR and had specificity only for the NDV-G-VII genotype. Additionally, this system was more rapid and had lower cost than real-time RT-PCR. Based on the results, the RT-LAMP-based assay is a useful tool for the rapid and sensitive diagnosis of NDV-G-VII infection.


Subject(s)
Newcastle Disease , Newcastle disease virus , Animals , Newcastle disease virus/genetics , Reverse Transcription , Newcastle Disease/diagnosis , Chickens/genetics , Genotype
4.
Saudi J Biol Sci ; 29(4): 3023-3032, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35531212

ABSTRACT

Knowledge of soil weed seed bank is important for population dynamics studied, establishment of appropriate weed management programs, a little effort in understanding weed seed bank can give valuable information about what weeds to expect in growing season, weed density, and when most weed germination will take place. In this study, a two - year's, two sites were carried out with the aim of assessing weed seed bank status of the soil throughout 2018 and 2019. A site was worked out in Sakha Agriculture Research farm act as a clay soil, Kafr El-Sheikh Governorate, Agriculture Research Center (ARC). Another site was worked out in El-Ismailia Agr; Res; farm act as sandy soil, El-Ismailia Governorate, ARC. At each site, soil samples were selected from nine different places as like three Zigzag shapes divided into three, six and nine sites, "W" to act the whole soil area (30 faddan in Sakha farm, and 15 faddan in El-Ismailia farm). The soil samples were taken from topsoil 0-10 cm depth with an auger (core) 10 cm diameter the soils without tillage and before sowing the summer crop. The result of present the study in two different stations and soils, revealed that the number of soil samples to estimate weed seed banks should be either six or nine sites; each sample weighted 0.50 Kg soil with zigzag shape act a direct seed extraction technique to able recognize the abundance of weed species into the soil and their seed density. The aim is to improve integrated weed control.

5.
J Biochem ; 170(3): 419-426, 2021 Oct 12.
Article in English | MEDLINE | ID: mdl-33950190

ABSTRACT

Doxorubicin is a chemotherapeutic drug widely utilized in cancer treatment. An enzyme critical to doxorubicin metabolism is the cytosolic sulfotransferase (SULT) SULT1C4. This study investigated the functional impact of SULT1C4 single nucleotide polymorphisms (SNPs) on the sulfation of doxorubicin by SULT1C4 allozymes. A comprehensive database search was performed to identify various SULT1C4 SNPs. Ten nonsynonymous SULT1C4 SNPs were selected, and the corresponding cDNAs, packaged in pGEX-2TK expression vector, were generated via site-directed mutagenesis. Respective SULT1C4 allozymes were bacterially expressed and purified by affinity chromatography. Purified SULT1C4 allozymes, in comparison with the wild-type enzyme, were analysed for sulphating activities towards doxorubicin and 4-nitrophenol, a prototype substrate. Results obtained showed clearly differential doxorubicin-sulphating activity of SULT1C4 allozymes, implying differential metabolism of doxorubicin through sulfation in individuals with distinct SULT1C4 genotypes.


Subject(s)
Doxorubicin/metabolism , Polymorphism, Single Nucleotide , Sulfotransferases/genetics , Sulfotransferases/metabolism , Cytosol/metabolism , Genotype , Humans , Isoenzymes/metabolism , Kinetics , Mutagenesis, Site-Directed , Nitrophenols/metabolism , Sulfates/metabolism
6.
Planta Med ; 87(6): 498-506, 2021 May.
Article in English | MEDLINE | ID: mdl-33572003

ABSTRACT

Radix Bupleuri is one of the most widely used herbal medicines in China for the treatment of fever, pain, and/or chronic inflammation. Quercitrin, epicatechin, and rutin, the flavonoids present in Radix Bupleuri, have been reported to display anti-inflammatory, antitumor, and antioxidant biological activities among others. Sulfation has been reported to play an important role in the metabolism of flavonoids. In this study, we aimed to systematically identify the human cytosolic sulfotransferase enzymes that are capable of catalyzing the sulfation of quercitrin, epicatechin, and rutin. Of the thirteen known human cytosolic sulfotransferases, three (cytosolic sulfotransferase 1A1, cytosolic sulfotransferase 1C2, and cytosolic sulfotransferase 1C4) displayed sulfating activity toward quercitrin, three (cytosolic sulfotransferase 1A1, cytosolic sulfotransferase 1A3, and cytosolic sulfotransferase 1C4) displayed sulfating activity toward epicatechin, and six (cytosolic sulfotransferase 1A1, cytosolic sulfotransferase 1A2, cytosolic sulfotransferase 1A3, cytosolic sulfotransferase 1B1, cytosolic sulfotransferase 1C4, and cytosolic sulfotransferase 1E1) displayed sulfating activity toward rutin. The kinetic parameters of the cytosolic sulfotransferases that showed the strongest sulfating activities were determined. To investigate the effects of genetic polymorphisms on the sulfation of quercitrin, epicatechin, and rutin, individual panels of cytosolic sulfotransferase allozymes previously prepared were analyzed and shown to display differential sulfating activities toward each of the three flavonoids. Taken together, these results provided a biochemical basis underlying the metabolism of quercitrin, epicatechin, and rutin through sulfation in humans.


Subject(s)
Catechin/chemistry , Quercetin/chemistry , Rutin/chemistry , Sulfotransferases/chemistry , China , Cytosol , Humans , Polymorphism, Genetic , Quercetin/analogs & derivatives , Sulfates , Sulfotransferases/genetics
7.
BMC Cancer ; 21(1): 155, 2021 Feb 12.
Article in English | MEDLINE | ID: mdl-33579235

ABSTRACT

BACKGROUND: p63, a member of the p53 gene family, is an important regulator for epithelial tissue growth and development. ∆Np63α is the main isoform of p63 and highly expressed in Non-melanoma skin cancer (NMSC). Extracellular signal-regulated kinase 3 (ERK3) is an atypical mitogen-activated protein kinase (MAPK) whose biochemical features and cellular regulation are distinct from those of conventional MAPKs such as ERK1/2. While ERK3 has been shown to be upregulated in lung cancers and head and neck cancers, in which it promotes cancer cell migration and invasion, little is known about the implication of ERK3 in NMSCs. METHODS: Fluorescent immunohistochemistry was performed to evaluate the expression levels of ΔNp63α and ERK3 in normal and NMSC specimens. Dunnett's test was performed to compare mean fluorescence intensity (MFI, indicator of expression levels) of p63 or ERK3 between normal cutaneous samples and NMSC samples. A mixed effects (ANOVA) test was used to determine the correlation between ΔNp63α and ERK3 expression levels (MFI). The regulation of ERK3 by ΔNp63α was studied by qRT-PCR, Western blot and luciferase assay. The effect of ERK3 regulation by ΔNp63α on cell migration was measured by performing trans-well migration assay. RESULTS: The expression level of ∆Np63α is upregulated in NMSCs compared to normal tissue. ERK3 level is significantly upregulated in AK and SCC in comparison to normal tissue and there is a strong positive correlation between ∆Np63α and ERK3 expression in normal skin and skin specimens of patients with AK, SCC or BCC. Further, we found that ∆Np63α positively regulates ERK3 transcript and protein levels in A431 and HaCaT skin cells, underlying the upregulation of ERK3 expression and its positive correlation with ∆Np63α in NMSCs. Moreover, similar to the effect of ∆Np63α depletion, silencing ERK3 greatly enhanced A431 cell migration. Restoration of ERK3 expression under the condition of silencing ∆Np63α counteracted the increase in cell migration induced by the depletion of ∆Np63α. Mechanistically, ERK3 inhibits the phosphorylation of Rac1 G-protein and the formation of filopodia of A431 skin SCC cells. CONCLUSIONS: ERK3 is positively regulated by ∆Np63α and mediates the role of ∆Np63α in suppressing cell migration in NMSC.


Subject(s)
Cell Movement , Gene Expression Regulation, Neoplastic , Mitogen-Activated Protein Kinase 6/metabolism , Skin Neoplasms/pathology , Transcription Factors/metabolism , Transcriptional Activation , Tumor Suppressor Proteins/metabolism , rac1 GTP-Binding Protein/metabolism , Cell Line , Cell Line, Tumor , Humans , Mitogen-Activated Protein Kinase 6/genetics , Phosphorylation , Skin Neoplasms/genetics , Skin Neoplasms/metabolism , Transcription Factors/genetics , Tumor Suppressor Proteins/genetics , rac1 GTP-Binding Protein/genetics
8.
Bioresour Technol ; 309: 123311, 2020 Aug.
Article in English | MEDLINE | ID: mdl-32289661

ABSTRACT

The present study investigated the possibility of valorizing rape straw through anaerobic digestion and the possibility of improving biomethane yield by pretreatment with H2SO4, combined H2SO4 with steam explosion (SE) and SE combined with superfine grinding (SFG). To evaluate the pretreatment method efficiency, several analytical techniques were applied. Additionally, the performance of co-digesting of cattle manure (CM) with pretreated rape straw (PRS) at different ratios was evaluated. The results showed that combined pretreatment could dissolve the lignocellulosic fiber structure, which positively stimulated methane yield. The highest cumulative CH4 yield (CMY) of 305.7 mLg-1VS was achieved by combined SE at 180 °C for 5 min with SFG, which was 77.84% higher than the untreated. The CMY was further improved by 11.4-59% higher than the control (CM) using co-digestion. This study confirmed that, under optimal parameters of AD, pretreatment with SEG180 could significantly boost the CMY from co-digestion of CM and PRS.


Subject(s)
Biofuels , Manure , Anaerobiosis , Animals , Cattle , Methane , Steam
9.
Pharmacogenet Genomics ; 29(5): 99-105, 2019 07.
Article in English | MEDLINE | ID: mdl-31145702

ABSTRACT

OBJECTIVES: Phenylephrine and salbutamol are drugs that are used widely to treat diseases/disorders, such as nasal congestion, hypotension, and asthma, in individuals of different age groups. Human cytosolic sulfotransferase (SULT) SULT1A3 has been shown to be critically involved in the metabolism of these therapeutic agents. This study was carried out to investigate the effects of single nucleotide polymorphisms of human SULT1A3 and SULT1A4 genes on the sulfation of phenylephrine and salbutamol by SULT1A3 allozymes. MATERIALS AND METHODS: Wild-type and SULT1A3 allozymes, prepared previously by site-directed mutagenesis in conjunction with bacterial expression and affinity purification, were analyzed for sulfating activity using an established assay procedure. RESULTS: Purified SULT1A3 allozymes, in comparison with the wild-type enzyme, showed differential sulfating activities toward phenylephrine and salbutamol. Kinetic studies showed further significant variations in their substrate-binding affinity and catalytic activity toward phenylephrine and salbutamol. CONCLUSION: The results obtained showed clearly the differential enzymatic characteristics of SULT1A3 allozymes in mediating the sulfation of phenylephrine and salbutamol. This information may contribute toward a better understanding of the pharmacokinetics of these two drugs in individuals with distinct SULT1A3 and/or SULT1A4 genotypes.


Subject(s)
Albuterol/metabolism , Arylsulfotransferase/genetics , Phenylephrine/metabolism , Sulfotransferases/genetics , Albuterol/therapeutic use , Arylsulfotransferase/chemistry , Arylsulfotransferase/metabolism , Asthma/drug therapy , Asthma/genetics , Genotype , Humans , Hypotension/drug therapy , Hypotension/genetics , Isoenzymes/genetics , Isoenzymes/metabolism , Kinetics , Mutagenesis, Site-Directed , Phenylephrine/therapeutic use , Polymorphism, Single Nucleotide/genetics , Sulfates/metabolism , Sulfotransferases/chemistry , Sulfotransferases/metabolism
10.
Pharmacol Rep ; 71(2): 257-265, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30822619

ABSTRACT

BACKGROUND: Non-opioid and opioid analgesics, as over-the-counter or prescribed medications, are widely used for the management of a diverse array of pathophysiological conditions. Previous studies have demonstrated the involvement of human cytosolic sulfotransferase (SULT) SULT1A1 in the sulfation of acetaminophen, O-desmethylnaproxen (O-DMN), and tapentadol. The current study was designed to investigate the impact of single nucleotide polymorphisms (SNPs) of the human SULT1A1 gene on the sulfation of these analgesic compounds by SULT1A1 allozymes. METHODS: Human SULT1A1 genotypes were identified by database search. cDNAs corresponding to nine SULT1A1 nonsynonymous missense coding SNPs (cSNPs) were generated by site-directed mutagenesis. Recombinant wild-type and SULT1A1 allozymes were bacterially expressed and affinity-purified. Purified SULT1A1 allozymes were analyzed for sulfation activity using an established assay procedure. RESULTS: Compared with the wild-type enzyme, SULT1A1 allozymes were shown to display differential sulfating activities toward three analgesic compounds, acetaminophen, O-desmethylnaproxen (O-DMN), and tapentadol, as well as the prototype substrate 4NP. CONCLUSION: Results obtained indicated clearly the impact of genetic polymorphisms on the drug-sulfation activity of SULT1A1 allozymes. Such information may contribute to a better understanding about the differential metabolism of acetaminophen, O-DMN, and tapentadol in individuals with different SULT1A1 genotypes.


Subject(s)
Acetaminophen/metabolism , Arylsulfotransferase/genetics , Naproxen/analogs & derivatives , Tapentadol/metabolism , Analgesics, Non-Narcotic/metabolism , Analgesics, Opioid/metabolism , Cytosol/metabolism , Escherichia coli/cytology , Genotype , Humans , Isoenzymes , Mutagenesis, Site-Directed , Naproxen/metabolism , Polymorphism, Single Nucleotide , Sulfates/metabolism
11.
BMC Public Health ; 16(1): 1113, 2016 10 22.
Article in English | MEDLINE | ID: mdl-27770781

ABSTRACT

BACKGROUND: Studies uncovering factors beyond socio-economic status (SES) that would explain racial and ethnic disparities in mortality are scarce. METHODS: Using prospective cohort data from the Third National Health and Nutrition Examination Survey (NHANES III), we examined all-cause and cause-specific mortality disparities by race, mediation through key factors and moderation by age (20-49 vs. 50+), sex and poverty status. Cox proportional hazards, discrete-time hazards and competing risk regression models were conducted (N = 16,573 participants, n = 4207 deaths, Median time = 170 months (1-217 months)). RESULTS: Age, sex and poverty income ratio-adjusted hazard rates were higher among Non-Hispanic Blacks (NHBs) vs. Non-Hispanic Whites (NHW). Within the above-poverty young men stratum where this association was the strongest, the socio-demographic-adjusted HR = 2.59, p < 0.001 was only partially attenuated by SES and other factors (full model HR = 2.08, p = 0.003). Income, education, diet quality, allostatic load and self-rated health, were among key mediators explaining NHB vs. NHW disparity in mortality. The Hispanic paradox was observed consistently among women above poverty (young and old). NHBs had higher CVD-related mortality risk compared to NHW which was explained by factors beyond SES. Those factors did not explain excess risk among NHB for neoplasm-related death (fully adjusted HR = 1.41, 95 % CI: 1.02-2.75, p = 0.044). Moreover, those factors explained the lower risk of neoplasm-related death among MA compared to NHW, while CVD-related mortality risk became lower among MA compared to NHW upon multivariate adjustment. CONCLUSIONS: In sum, racial/ethnic disparities in all-cause and cause-specific mortality (particularly cardiovascular and neoplasms) were partly explained by socio-demographic, SES, health-related and dietary factors, and differentially by age, sex and poverty strata.


Subject(s)
Cardiovascular Diseases/mortality , Ethnicity , Health Status Disparities , Neoplasms/mortality , Poverty , Racial Groups , Social Class , Adult , Aged , Allostasis , Cause of Death , Diet , Educational Status , Female , Humans , Income , Male , Middle Aged , Nutrition Surveys , Proportional Hazards Models , Prospective Studies , Risk , United States/epidemiology , Young Adult
12.
Endocrinology ; 156(4): 1441-52, 2015 Apr.
Article in English | MEDLINE | ID: mdl-25643156

ABSTRACT

Polycystic ovary syndrome (PCOS) is associated with reproductive, endocrine, and metabolic abnormalities. Because hyperandrogenism is the most consistent PCOS feature, we used wild-type (WT) and androgen receptor (AR) knockout (ARKO) mice, together with a mouse model of PCOS, to investigate the contribution of genomic AR-mediated actions in the development of PCOS traits. PCOS features were induced by prenatal exposure to dihydrotestosterone (250 µg) or oil vehicle (control) on days 16-18 of gestation in WT, heterozygote, and homozygote ARKO mice. DHT treatment of WT mice induced ovarian cysts (100% vs 0%), disrupted estrous cycles (42% vs 100% cycling), and led to fewer corpora lutea (5.0±0.4 vs 9.8±1.8). However, diestrus serum LH and FSH, and estradiol-induced-negative feedback as well as hypothalamic expression of kisspeptin, neurokinin B, and dynorphin, were unaffected by DHT treatment in WT mice. DHT-treated WT mice exhibited a more than 48% increase in adipocyte area but without changes in body fat. In contrast, heterozygous and homozygous ARKO mice exposed to DHT maintained comparable ovarian (histo)morphology, estrous cycling, and corpora lutea numbers, without any increase in adipocyte size. These findings provide strong evidence that genomic AR signaling is an important mediator in the development of these PCOS traits with a dose dependency that allows even AR haplosufficiency to prevent induction by prenatal androgenization of PCOS features in adult life.


Subject(s)
Hyperandrogenism/prevention & control , Polycystic Ovary Syndrome/prevention & control , Prenatal Exposure Delayed Effects/prevention & control , Receptors, Androgen/metabolism , Adipose Tissue/metabolism , Androgens , Animals , Disease Models, Animal , Estrous Cycle , Female , Hyperandrogenism/chemically induced , Hyperandrogenism/metabolism , Mice , Mice, Knockout , Ovary/metabolism , Polycystic Ovary Syndrome/chemically induced , Polycystic Ovary Syndrome/metabolism , Pregnancy , Prenatal Exposure Delayed Effects/metabolism , Receptors, Androgen/genetics
13.
Appl Radiat Isot ; 89: 13-7, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24561650

ABSTRACT

Aqueous solutions of the organic dye Rhodamine B were found to be useful in measuring radiation doses in the range 0.1-2 kGy. Either spectrophotometric or spectrofluorometric measurements can be used. The dosimeter readout is unchanged for at least 60 days after irradiation if the solution is kept in the dark at room temperature. Effects of pH and the dye concentration on the dosimeter response were investigated. Radiation chemical yield and fluorescence quantum yield were also calculated.


Subject(s)
Gamma Rays , Radiometry/methods , Rhodamines/radiation effects , Spectrometry, Fluorescence , Spectrophotometry, Ultraviolet
14.
Virus Genes ; 48(1): 153-9, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24258394

ABSTRACT

The dahlia (Dahlia variabilis) genome contains an endogenous pararetrovirus sequence (EPRS) tentatively designated as DvEPRS. The DvEPRS shares genome structure and organization that is typical of members of the Caulimovirus genus. Studies were carried out to better understand the nature of this integration and to determine the gene expression of this DvEPRS. Genomic Southern hybridization showed multiple and random integration events of the DvEPRS in the dahlia genome. To investigate the presence of DvEPRS transcripts, RT-PCR was done on DNase-treated total RNA from DvEPRS-infected dahlia plants. Results showed the expression of open reading frames I, V, and VI. Direct PCR from sap extracts produced more intense DNA amplicons of Dahlia mosaic virus and Dahlia common mosaic virus which are believed to exist as typical episomal caulimoviruses, whereas significantly less intense amplicon was seen in case of DvEPRS in comparison with internal transcribed spacer region of dahlias amplicon. The DvEPRS in wild and cultivated species of Dahlia offer a model system to study the molecular events underlying the ecology, evolution and spread of DvEPRS within natural and managed ecosystems and the factors affecting integration of these EPRS in the plant genome.


Subject(s)
Caulimovirus/physiology , Dahlia/virology , Gene Expression , Viral Proteins/biosynthesis , Virus Integration , Blotting, Southern , Caulimovirus/genetics , Genome, Plant , Viral Proteins/genetics
15.
Virus Genes ; 48(1): 140-52, 2014 Feb.
Article in English | MEDLINE | ID: mdl-24353027

ABSTRACT

Two distinct caulimoviruses, Dahlia mosaic virus (DMV) and Dahlia common mosaic virus, and an endogenous plant pararetroviral sequence (DvEPRS) were reported in Dahlia spp. DvEPRS, previously referred to as DMV-D10, was originally identified in the US from the cultivated Dahlia variabilis, and has also been found in New Zealand, Lithuania and Egypt, as well as in wild dahlia species growing in their natural habitats in Mexico. Sequence analysis of three new EPRSs from cultivated dahlias from Lithuania [D10-LT; 7,159 nucleotide level (nt)], New Zealand (D10-NZ, 7,156 nt), and the wild species, Dahlia rupicola, from Mexico (D10-DR, 7,133 nt) is reported in this study. The three EPRSs have the structure and organization typical of a caulimovirus species and showed identities among various open reading frames (ORFs) ranging between 71 and 97 % at the nt when compared to those or the known DvEPRS from the US. Examination of a dataset of seven full-length EPRSs obtained to date from cultivated and wild Dahlia spp. provided clues into genetic diversity of these EPRSs from diverse sources of dahlia. Phylogenetic analyses, mutation frequencies, potential recombination events, selection, and fitness were evaluated as evolutionary evidences for genetic variation. Assessment of all ORFs using phylogenomic and population genetics approaches suggests a wide genetic diversity of EPRSs occurring in dahlias. Phylogenetic analyses show that the EPRSs from various sources form one clade indicating a lack of clustering by geographical origin. Grouping of various EPRSs into two host taxa (cultivated vs. wild) shows little divergence with respect to their origin. Population genetic parameters demonstrate negative selection for all ORFs, with the reverse transcriptase region more variable than other ORFs. Recombination events were found which provide evolutionary evidence for genetic diversity among dahlia-associated EPRSs. This study contributes to an increased understanding of molecular population genetics and evolutionary pathways of these reverse transcribing viral elements.


Subject(s)
Caulimovirus/classification , Caulimovirus/isolation & purification , Dahlia/virology , Caulimovirus/genetics , Cluster Analysis , Gene Order , Genes, Viral , Genetic Variation , Lithuania , Mexico , Molecular Sequence Data , New Zealand , Open Reading Frames , Phylogeny , RNA, Viral/genetics , Sequence Analysis, DNA , Sequence Homology, Amino Acid
16.
J Biol Regul Homeost Agents ; 27(3): 693-703, 2013.
Article in English | MEDLINE | ID: mdl-24152838

ABSTRACT

Diabetic nephropathy (DN) is one of the most serious complications of type I and type II diabetes. DN is characterized by hyperfiltration, hypertrophy, extracellular matrix accumulation, and proteinuria. This advances into renal fibrosis and loss of renal function. Reactive oxygen species (ROS) and TGF-beta have been implicated in the pathogenesis of diabetic nephropathy. Early stages of diabetic nephropathy are also associated with alterations in renal sodium handling as well as hypertension; both are processes linked by involvement of the arachidonic acid (AA) metabolites, 20-hydroxyeicosatetraenoic acid (20-HETE, produced by cytochrome P450-4a, (CYP4A) and epoxyeicosatrienoic acids (EETs). Indeed, metabolism of AA is increased in a rat model of diabetes. In this study, we demonstrate that rats with streptozotocin-induced diabetes of 1 month duration develop renal hypertrophy and increased fibronectin and TGF-beta1 expression/cortical levels concomitant with an increase in CYP4A expression and 20 HETE production. These results were also paralleled by an increase in reactive oxygen species (ROS) production and NADPH oxidase activity. Treatment of diabetic rats with HET0016, selective inhibitor of CYP 4A, prevented all these changes. Our results suggest that diabetes-induced induction of CYP4A and 20-HETE production could be a major pathophysiological mechanism leading to activation of ROS through an NADPH dependent pathway and TGF-beta1 thus resulting in major renal pathology. Inhibitors of 20-HETE production could thus have an important therapeutic potential in the treatment of diabetic nephropathy.


Subject(s)
Cytochrome P-450 CYP4A/physiology , Diabetes Mellitus, Experimental/complications , Diabetic Nephropathies/etiology , Hydroxyeicosatetraenoic Acids/physiology , Kidney/enzymology , Animals , Kidney/pathology , Male , NADPH Oxidases/metabolism , Rats , Reactive Oxygen Species/metabolism , Streptozocin , Transforming Growth Factor beta1/biosynthesis
17.
Hum Reprod ; 28(2): 306-14, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23203216

ABSTRACT

STUDY QUESTION: Does the selection of sperm for ICSI based on their ability to bind to hyaluronan improve the clinical pregnancy rates (CPR) (primary end-point), implantation (IR) and pregnancy loss rates (PLR)? SUMMARY ANSWER: In couples where ≤ 65% of sperm bound hyaluronan, the selection of hyaluronan-bound (HB) sperm for ICSI led to a statistically significant reduction in PLR. WHAT IS KNOWN AND WHAT THIS PAPER ADDS: HB sperm demonstrate enhanced developmental parameters which have been associated with successful fertilization and embryogenesis. Sperm selected for ICSI using a liquid source of hyaluronan achieved an improvement in IR. A pilot study by the primary author demonstrated that the use of HB sperm in ICSI was associated with improved CPR. The current study represents the single largest prospective, multicenter, double-blinded and randomized controlled trial to evaluate the use of hyaluronan in the selection of sperm for ICSI. DESIGN: Using the hyaluronan binding assay, an HB score was determined for the fresh or initial (I-HB) and processed or final semen specimen (F-HB). Patients were classified as >65% or ≤ 65% I-HB and stratified accordingly. Patients with I-HB scores ≤ 65% were randomized into control and HB selection (HYAL) groups whereas patients with I-HB >65% were randomized to non-participatory (NP), control or HYAL groups, in a ratio of 2:1:1. The NP group was included in the >65% study arm to balance the higher prevalence of patients with I-HB scores >65%. In the control group, oocytes received sperm selected via the conventional assessment of motility and morphology. In the HYAL group, HB sperm meeting the same visual criteria were selected for injection. Patient participants and clinical care providers were blinded to group assignment. PARTICIPANTS AND SETTING: Eight hundred two couples treated with ICSI in 10 private and hospital-based IVF programs were enrolled in this study. Of the 484 patients stratified to the I-HB > 65% arm, 115 participants were randomized to the control group, 122 participants were randomized to the HYAL group and 247 participants were randomized to the NP group. Of the 318 patients stratified to the I-HB ≤ 65% arm, 164 participants were randomized to the control group and 154 participants were randomized to the HYAL group. MAIN RESULTS AND THE ROLE OF CHANCE: HYAL patients with an F-HB score ≤ 65% demonstrated an IR of 37.4% compared with 30.7% for control [n = 63, 58, P > 0.05, (95% CI of the difference -7.7 to 21.3)]. In addition, the CPR associated with patients randomized to the HYAL group was 50.8% when compared with 37.9% for those randomized to the control group (n = 63, 58, P > 0.05). The 12.9% difference was associated with a risk ratio (RR) of 1.340 (RR 95% CI 0.89-2.0). HYAL patients with I-HB and F-HB scores ≤ 65% revealed a statistically significant reduction in their PLR (I-HB: 3.3 versus 15.1%, n = 73, 60, P = 0.021, RR of 0.22 (RR 95% CI 0.05-0.96) (F-HB: 0.0%, 18.5%, n = 27, 32, P = 0.016, RR not applicable due to 0.0% value) over control patients. The study was originally planned to have 200 participants per arm providing 86.1% power to detect an increase in CPR from 35 to 50% at α = 0.05 but was stopped early for financial reasons. As a pilot study had demonstrated that sperm preparation protocols may increase the HB score, the design of the current study incorporated a priori collection and analysis of the data by both the I-HB and the F-HB scores. Analysis by both the I-HB and F-HB score acknowledged the potential impact of sperm preparation protocols. BIAS, CONFOUNDING AND OTHER REASONS FOR CAUTION: Selection bias was controlled by randomization. Geographic and seasonal bias was controlled by recruiting from 10 geographically unique sites and by sampling over a 2-year period. The potential for population effect was controlled by adjusting for higher prevalence rates of >65% I-HB that naturally occur by adding the NP arm and to concurrently recruit >65% and ≤ 65% I-HB subjects. Monitoring and site audits occurred regularly to ensure standardization of data collection, adherence to the study protocol and subject recruitment. Subgroup analysis based on the F-HB score was envisaged in the study design. GENERALIZABILITY TO OTHER POPULATIONS: The study included clinics using different sperm preparation methods, located in different regions of the USA and proceeded in every month of the year. Therefore, the results are widely applicable.


Subject(s)
Hyaluronic Acid/metabolism , Semen Analysis/methods , Spermatozoa/metabolism , Adult , Birth Rate , Double-Blind Method , Embryo Implantation , Female , Humans , Male , Pregnancy , Pregnancy Rate , Sperm Injections, Intracytoplasmic/methods , Spermatozoa/physiology
18.
Arch Virol ; 156(11): 2079-84, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21837417

ABSTRACT

The genome structure and organization of endogenous caulimovirus sequences from dahlia (Dahlia spp), dahlia mosaic virus (DMV)-D10 from three wild species, D. coccinea (D10-DC), D. sherffii (D10-DS) and D. tenuicaulis (D10-DT), were determined and compared to those from cultivated species of dahlia, D. variabilis (DvEPRS). The complete ca. 7-kb dsDNA genomes of D10-DC, D10-DS, and D10-DT had a structure and organization typical of a caulimovirus and shared 89.3 to 96.6% amino acid sequence identity in various open reading frames (ORF) when compared to DvEPRS. The absence of the aphid transmission factor and the truncated coat protein fused with the reverse transcriptase ORF were common among these DMV-D10 isolates from wild Dahlia species.


Subject(s)
Caulimovirus/genetics , Dahlia/virology , Genome, Viral , Plant Diseases/virology , Base Sequence , Caulimovirus/chemistry , Caulimovirus/classification , Caulimovirus/isolation & purification , Ecosystem , Genomics , Molecular Sequence Data , Phylogeny , Sequence Alignment
19.
Life Sci ; 89(13-14): 430-8, 2011 Sep 26.
Article in English | MEDLINE | ID: mdl-21777594

ABSTRACT

AIM: The purpose of this study was to develop transdermal films (TFs) with the addition of different polymer ratios that incorporated 0.5% tenoxicam in order to ensure maximum controlled and sustained drug release capacity. Tenoxicam is a non-steroidal anti-inflammatory drug (NSAID) widely used in the treatment of rheumatic diseases and characterized by its efficacy and reduced side effects in comparison to other NSAIDs. MAIN METHODS: Transdermal films of tenoxicam were designed with the Eudragit L30D-55copolymer with permeation enhancers like polyethylene glycol (PEG) and propylene glycol (PG) incorporated at different concentrations using the casting evaporation technique. Evaluations of these formulae were performed through mechanical characterizations and Fourier Transform Infrared Spectroscopy [FTIR]. In-vitro release studies were performed during 24h using diffusion cells. The film formulations with optimum in vitro-release rate have been taken up for testing of the anti-inflammatory effects and the sustaining action of tenoxicam. The in-vivo studies performed included carrageenan-induced hind paw edema and skin biopsies in Wistar rats. KEY FINDINGS: Formulation (F7) had the best effective combination [glycerol (0.25g), PEG200 (0.5g), PEG400 (1g) and PG (10%) and 0.5% dispersed drug] among all of the tenoxicam TF formulations studied. Also, this formula had the highest release value than formula 1 (F1) that contains [glycerol (2.5g), PEG200 (0.5g), PEG400 (0.5g) and 0.5% dissolved drug] or a commercially available gel after 24h. FTIR revealed that there was an interaction between the polymer and the drug. The drug-polymer interaction occurring between tenoxicam and Eudragit L30D-55 seems to cause a drag effect, leading to a delay of the tenoxicam release from the Eudragit L film. SIGNIFICANCE: When the films were applied half an hour before the subplantar injection of carrageenan in the hind paw of Wistar rats, it was observed that formula F7 provided maximum inhibition of paw edema in rats over 24h in contrast to both formula F1 and the marketed piroxicam gel as a reference. This was also confirmed histopathologically from skin biopsies. Thus, we show that tenoxicam can be formulated into transdermal films to sustain its release characteristics, and the polymeric composition of PEG200/PEG400 at a ratio of 1:2 and 10% PG was found to be the best choice to manufacture tenoxicam TF.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/administration & dosage , Edema/drug therapy , Piroxicam/analogs & derivatives , Skin/drug effects , Transdermal Patch , Animals , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Edema/chemically induced , Edema/pathology , Lower Extremity/pathology , Male , Methacrylates/chemistry , Piroxicam/administration & dosage , Piroxicam/therapeutic use , Polyethylene Glycols/chemistry , Polymers/chemistry , Propylene Glycol/chemistry , Rats , Rats, Wistar , Skin/pathology , Transdermal Patch/trends
20.
Handb Exp Pharmacol ; (194): 261-81, 2009.
Article in English | MEDLINE | ID: mdl-19655110

ABSTRACT

The somatosensory effects of natural products such as capsaicin, mustard oil, and menthol have been long recognized. Over the last decade, the identification of transient receptor potential (TRP) channels in primary sensory neurons as the targets for these agents has led to an explosion of research into the roles of "thermoTRPs" TRPV1, TRPV2, TRPV3, TRPV4, TRPA1, and TRPM8 in nociception. In concert, through the efforts of many industrial and academic teams, a number of agonists and antagonists of these channels have been discovered, paving the way for a better understanding of sensory biology and, potentially, for novel treatments for diseases.


Subject(s)
Pain/metabolism , Sensory Receptor Cells/metabolism , Transient Receptor Potential Channels/metabolism , Analgesics/pharmacology , Animals , Humans , Pain/drug therapy , Pain/physiopathology , Pain Threshold , Sensory Receptor Cells/drug effects , TRPM Cation Channels/metabolism , TRPV Cation Channels/metabolism , Transient Receptor Potential Channels/drug effects
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