Bacillus thuringiensis insecticidal delta-endotoxin: diversity of crystal proteins and its relatedness to the toxicity spectrum

Bacillus thuringiensis strains produce crystal delta-endotoxins which exhibit a diverse toxicity spectrum. In order to explore the basis of toxin specificity, a comparison of the activity of 13 strains belonging to seven serotypes was made against three insect species. The delta-endotoxin crystals were purified and their polypeptide composition analyzed by SDS-PAGE. Among the strains studied, the delta-endotoxins consist of a variety of crystal proteins in the 60-144 KDa size range. On the basis of molecular mass, endotoxins maybe grouped into two classes; one contained both high (125-144 KDa, P1) and medium sized (60-66 KDa, P2) proteins and a second class consisting of only the high Mr polypeptides. Immunoblotting with B. aizawai P1 antiserum revealed antigenic cross-reaction with one or more of the polypeptides in 125-144 KDa range in all the strains studied. When the crystal proteins from different strains were immunoblotted with kurstaki P2 antiserum, none of the P1 protein crossreacted suggesting that the P1 and P2 proteins are not structurally related. However, the B. kurstaki P2 antiserum crossreacted with 66 KDa proteins in some other strains which underlines a structural homology in this class of the toxic polypeptides. Toxicity studies revealed that the high Mr P1 proteins of all the strains in this study were active against lepidopteran (Pieris brassicae and Diacrisia obliqua) larvae. B. thuringiensis aizawai strains exhibit a dual toxicity associated with the high Mr (130-135 KDa; P1) proteins. The P2 crystal proteins (60-66 KDa) also showed dual toxicity against the lepidopteran and dipteran larvae but were found to be structurally and immunologically distinct.     

Isolation of an entomopoxvirus and characterization of its DNA.

An entomopoxvirus from Choristoneura biennis was propagated in the eastern spruce budworm, Choristoneura fumiferana. The virus, released from its inclusion bodies by alkaline treatment, sedimented as a homogeneous band by rate zonal centrifugation through sucrose gradients. Electron microscopic examination showed that the particles possessed the mulberrylike structure that is typical of intact entomopox virions. The virus also produced a single band by isopycnic centrifugation in CsCl at a density of 1.31 g/ml. The genome, extracted with sodium lauryl sarcosinate and Pronase, sedimented as a single band through sucrose gradients, and its MW was calculated to be 132?142 × 10⁶ when compared to marker DNA from coliphage T2. Its sedimentation coefficient was 59S. The G + C content of the viral genome was investigated by thermal denaturation and buoyant density analysis in CsCl. The DNA melted at 61.6° and had a density of 1.6849 g/ml. These values are equivalent to a G + C content of 26.6% and 25.4%, respectively, which are considerably lower than those reported for the genomes of vertebrate poxviruses (G + C, 32.5–39%). The difference in G + C content between vertebrate and invertebrate poxviruses was confirmed by characterizing the genome of a second entomopoxvirus with respect to its t_m and buoyant density. The double-stranded nature of the DNA from invertebrate poxviruses was established from melting profiles and reaction with formaldehyde.     

Depression,sleeping pattern,and suicidal ideation among medical students in Bangladesh: a cross-sectional pilot study

Journal of Public Health - Depression is a major morbidity and the most common mental disorder among the medical students in medical schools globally. Undergraduate students suffer stress more due...     

Pakistan’s Response to COVID-19: Overcoming National and International Hypes to Fight the Pandemic

The COVID-19 outbreak started as pneumonia in December 2019 in Wuhan, China. The subsequent pandemic was declared as the sixth public health emergency of international concern on January 30, 2020, by the World Health Organization. Pakistan could be a potential hotspot for COVID-19 owing to its high population of 204.65 million and its struggling health care and economic systems. Pakistan was able to tackle the challenge with relatively mild repercussions. The present analysis has been conducted to highlight the situation of the disease in Pakistan in 2020 and the measures taken by various stakeholders coupled with support from the community to abate the risk of catastrophic spread of the virus.     

Prevalence of Vision Loss and Associations With Age-Related Eye Diseases Among Nursing Home Residents Aged ≥65 Years

ObjectivesTo examine data from Delaware nursing homes to determine prevalence of age-related eye diseases (AREDs), vision impairment, and blindness and to compare the findings with the results of 11 US investigations of vision and eye health in nursing homes.DesignThis is a cross-sectional, retrospective study of nursing home patients.Setting and ParticipantsTwenty nursing homes in Delaware participated in the study, yielding comprehensive eye examination records for 2019 study participants.MethodsSummary statistics and regression analyses.ResultsThe overall prevalence of vision impairment or blindness was 63.8% and was above 60% for each age, sex, and race category. Prevalence of vision impairment or blindness was 68.4% among patients with cataracts, 69.4% among patients with macular degeneration, 70.5% among patients with glaucoma, and 68.4% among patients with diabetic retinopathy. Prevalence of blindness was 14.1%. Among patients with AREDs, prevalence of blindness ranged from 15.0% for patients with cataracts to 22.6% for patients with diabetic retinopathy. When compared with other investigations, we found wide variation in vision and eye factors reported and wide variation in the prevalence of those factors. Only 4 studies diagnosed both AREDs and visual function. Seven studies reported AREDs, and 7 reported vision impairment and/or blindness. Vision impairment or blindness ranged from 29% to 67%; cataract ranged from 32% to 83%; macular degeneration ranged from 4.6% to 70.7%. Glaucoma ranged from 5.3% to 41.4%; diabetic retinopathy ranged from 1.7% to 3.1%.Conclusions and ImplicationsComprehensive eye examinations showed that vision impairment and blindness affected 63.8% of nursing home residents. Compared with other studies, there was a wide range of vision factors reported and wide variation in the prevalence of vision impairment or blindness and AREDs. This investigation suggests the importance of eye care in nursing homes and the importance of reporting standard vision and eye health factors to inform policy and practice.     

Impact of United States 2017 Immigration Policy changes on missed appointments at two Massachusetts Safety-Net Hospitals

Introduction

Studies have shown mixed findings regarding the impact of immigration policy changes on immigrants’ utilization of primary care.

Methods

We used a difference-in-differences analysis to compare changes in missed primary care appointments over time across two groups: patients who received care in Spanish, Portuguese, or Haitian Creole, and non-Hispanic, white patients who received care in English.

Results

After adjustment for age, sex, race, insurance, hospital system, and presence of chronic conditions, immigration policy changes were associated with an absolute increase in the missed appointment prevalence of 0.74 percentage points (95% confidence interval: 0.34, 1.15) among Spanish, Portuguese and Haitian-Creole speakers. We estimated that missed appointments due to immigration policy changes resulted in lost revenue of over $185,000.

Conclusions

We conclude that immigration policy changes were associated with a significant increase in missed appointments among patients who receive medical care in languages other than English.

     

Effects of dietary supplementation of N-acetylcysteine on cigarette smoke-related DNA adducts in rat tissues

Cigarette smoking plays a major role in the etiology of several human cancers. It is believed that formation of DNA adducts is an initial step in the carcinogenic process. In this study, we have examined the ability of dietary N-acetylcysteine (NAG) to inhibit the formation of cigarette smoke-related DNA adducts in various tissues of rats. Female Sprague-Dawley rats were exposed to cigarette smoke (10 mg TPM/m(3)) in a whole-body exposure chamber for 6 h per day, seven days a week for four weeks. The smoke-exposed groups were provided either an unrefined diet or diets supplemented with low (5,000 ppm) or high (20,000 ppm) dose of NAG. A sham group was given control diet and maintained on filtered ambient air. Tissue DNA analysis of smoke-exposed rats by nuclease P1-version of the P-32-postlabeling assay showed up to 6 adducts in the following descending order expressed as total adducts/10(10) nucleotides: 1 predominant (no. 5) and 4 (no. 1-no. 4) minor adducts in the (219 +/- 36), 6 minor adducts in the heart (93 +/- 11), 5 adducts in the trachea (50 +/- 16), and 4 adducts in the bladder (50 +/- 3.5); sham-treated animals showed 2 or 3 adducts in each tissue but at 4-20-fold lower levels. Dietary intervention with either high or low dose of NAC did not affect the levels of most adducts, except for the following: a 30-40% increase (P<0.05) for adducts 3 and 4 in the lung; a 40-50% decrease (P<0.05) for adduct 2 in the trachea; and a 30% increase (P<0.05) for adduct 2 in the bladder. In a second experiment conducted under identical conditions, most major and minor adducts remained unaffected with NAC intervention, except for adduct 2 in the trachea which was somewhat diminished. These results suggest that dietary NAC intervention does not significantly influence the levels of most major and minor adducts. However, some minor adducts in the lung, trachea and bladder were modulated differentially.     

A comparative study of allometric scaling with plasma concentrations predicted by species-invariant time methods

The objective of this study is to compare the empirical allometric approaches with species invariant time methods using equivalent time, kallynochron, apolysichron, and dienetichrons. Pharmacokinetic parameters (clearance, volume of distribution, and elimination half-life) of ethosuximide, cyclosporine and ciprofloxacin were scaled-up from animal data obtained from the literature. Two methods were utilized to generate plots for the prediction of clearance in humans: (i) clearance versus body weight (simple allometric equation); and (ii) the product of clearance and maximum life-span potential (MLP) versus body weight. Plasma concentrations of each of the drugs were predicted using elementary and complex Dedrick plots, equivalent time with an exponent of 0.25 and equivalent time with the exponent obtained from the plot of body weight and half-life. Plasma concentrations of cyclosporine and ciprofloxacin were also predicted by MLP normalization (dienetichrons). Almost similar results in the pharmacokinetic parameters of the tested drugs were obtained by the allometric approach and by the species invariant time methods.     

Inhibition of cigarette smoke-related lipophilic DNA adducts in rat tissues by dietary oltipraz

The present study investigated the effects of dietary oltipraz on cigarette smoke-related lipophilic DNA adduct formation. Female Sprague- Dawley rats were exposed daily to sidestream cigarette smoke in a whole- body exposure chamber 6 h/day for 4 consecutive weeks. One group of rats was maintained on control diet while another group received the same diet supplemented with either a low (167 p.p.m.) or high (500 p.p.m.) dose of oltipraz, starting 1 week prior to initiation of smoke exposure until the end of the experiment. Analysis of lipophilic DNA adducts by the nuclease P1-mediated 32P-post-labeling showed up to five smoke-related adducts. Adduct no. 5 predominated in both the lung and the heart while adduct nos 3 and 2 predominated in the trachea and bladder, respectively. Quantitative analysis revealed that the total adduct level was the highest in lungs (270+/-68 adducts/10(10) nucleotides), followed by trachea (196+/-48 adducts/10(10) nucleotides), heart (141+/-22 adducts/10(10) nucleotides) and bladder (85+/-16 adducts/10(10) nucleotides). High dose oltipraz treatment reduced the adduct levels in lungs and bladder by >60%, while the reduction in lungs in the low-dose group was approximately 35%. In trachea, the effect of low and high dietary oltipraz on smoke DNA adduction was equivocal, while smoke-related DNA adducts in the heart were minimally inhibited by high-dose oltipraz. In a repeat experiment that employed a 3-fold lower dose of cigarette smoke, oltipraz (500 p.p.m.) was found to inhibit the formation of DNA adducts in rat lungs and trachea by 80 and 65%, respectively. These data clearly demonstrate a high efficacy of oltipraz in inhibiting the formation of cigarette smoke-induced DNA adducts in the target tissues.