Benzaldehyde-induced developmental genotoxicity triggers both neural and non-neuronal cells including the cells of immunity in Drosophila melanogaster.

Although benzaldehyde, an aromatic aldehyde, has been declared safe for uses in food, conflicting reports exist regarding its genotoxic and cytotoxic potentials in organisms. Our present study is the first attempt to evaluate the effects of exposure of benzaldehyde on the entire course of development of a eukaryote model organism, Drosophila melanogaster. Total time required for the initial appearance of the third instar larvae, pupae and adults increased dose dependently with the increasing dietary concentration of benzaldehyde. Exposure of flies to each concentration of benzaldehyde caused dose-dependent and significant reductions in the population of pupae and young adults of the fly. Developmental inhibition was associated with dose dependent and significant structural aberrations of larval polytene chromosomes like ectopic pairing, inversion, fusion, etc., and deformities of hemocytes and neuroblasts and death of hemocytes. As much as 34% (SD ± 1.76)-52% (SD ± 1.7) and 18% (SD ± 2.5)-40% (SD ± 3.38) hemocytes and neuroblasts, respectively, underwent nuclear deformations in response to dietary exposures of flies to BA 100-1000 mg/l. Moreover, 16% (SD ± 0.52)-31% (SD ± 1.97) and 19% (SD ± 0.3)-33% (SD ± 1.78) hemocyte mortalities in response to BA 100-1000 mg/l were determined by two cell viability assays. Thus our study revealed that benzaldehyde was genotoxic to Drosophila melanogaster larvae that might be responsible for larval cell death and their subsequent developmental retardation. As this fly possesses substantial genetic homology with human, possibility of developmental inhibition of the later due to exposure of this chemical during pregnancy may not be ruled out.

Silver nanoparticle-induced developmental inhibition of Drosophila melanogaster accompanies disruption of genetic material of larval neural stem cells and non-neuronal cells.

A few studies had determined the effects of silver nanoparticles on the development of Drosophila melanogaster. However, none had addressed its genotoxic effects on specific larval cells of the fly in details. This study was conducted to determine the effects of silver nanoparticle on the development of D. melanogaster with simultaneous evaluation of its genotoxic potential on specific larval cell types that play important roles in immunological defenses as well as growth and development. Five male and five female flies were maintained in standard Drosophila melanogaster culture medium containing varying concentrations of silver nanoparticles, i.e., 25, 50, 100, 200, and 300 mg/l with control culture medium containing no nanoparticle. Total time needed for stage-specific development, population yield, and genotoxic effects on third instar larval polytene chromosomes, hemocytes, and neuroblasts was determined. Body pigmentation of pupae and young adults was examined visually. In comparison with control, silver nanoparticles dose dependently inhibited the metamororphosis and population yields of pupae and young adults of Drosophila melanogaster. Every concentration of the nanoparticles inhibited pupa to adult conversion, with huge reduction under the influence of nanoparticle concentration of 100 mg/ml and above. Developmental inhibition was accompanied by dose-dependent and significant structural aberrations of larval polytene chromosomes and deformities of hemocytes and neuroblasts. Pupae and young adults also exhibited gradual discoloration of body with the increase in exposure to nanoparticle concentration.

Hydroxyurea responses in clinically varied beta, HbE-beta thalassaemia and sickle cell anaemia patients of Eastern India.

The haematological and clinical response to hydroxyurea was estimated in HbE-beta, beta thalassaemia and sickle cell anaemia patients of Eastern India, with variable clinical severity and transfusion requirement to determine whether hydroxyurea can help these patients to maintain their steady haemoglobin level without blood transfusions. Three hundred patients (189 HbE-beta thalassaemia, 95 beta thalassaemia and 16 other haemoglobinopathies including sickle cell anaemia) were selected for hydroxyurea therapy and were followed up for 48-60 months. Results suggest significant response to hydroxyurea therapy in 19 beta and 99 HbE-beta patients in the transfusion-dependent group (GR-I). All of them became transfusion-independent while on hydroxyurea therapy. The majority of responding patients were IVS1-5(G-C) in one of their alleles in HbE-beta cases (83 out of 119). Though IVS1-5(G-C) was found to be the commonest mutation in our selected patients, the mutational background of the patients does not found to have any significant correlation with the response category towards hydroxyurea as per the results observed in our study. But, the drug works pretty well in most of the transfusion-dependent patients, as these patients were withdrawn from regular blood transfusion. At the same time, partial or no response to the drug hydroxyurea was also recorded in our study.

Mycobacterium tuberculosis and non-tubercular mycobacterium infection in women with unexplained infertility from eastern India.

BACKGROUND: Genital tuberculosis (GTB) is an important cause of female infertility, especially in developing countries. The positive results of polymerase chain reaction (PCR) in endometrial GTB in the absence of tubal damage raise the possibility of the detection of sub-clinical or latent disease, with doubtful benefits of treatment. OBJECTIVE: This study aims to evaluate the Mycobacterium tuberculosis (MTB) and Non-tubercular Mycobacterium (NTM) infection by using Real-PCR technique in the menstrual blood samples of 120 unexplained infertile women. MATERIALS AND METHODS: In this cross-sectional study, 120 infertile women with unexplained infertility aged 20-35 yr old and normal hysterosalpingography findings were taken. Menstrual blood in the first 12 hr of menstruation containing the endometrial tissues from each participant was tested for MTB and NTM by Real-Time PCR. RESULTS: Among the selected 120 patients, only two were found to be positive for MTB infection. All remaining participants were negative for MTB infection. All participants were negative for NTM infection at the endometrium. CONCLUSION: Although, studies have indicated that PCR is a useful method in diagnosing early GTB disease in infertile women with no demonstrable evidence of tubal or endometrial involvement, our study showed that GTB is not the major problem in women with unexplained infertility.

Impacts of dietary exposure to sodium or potassium salts of nitrate and nitrite on the development of Drosophila melanogaster.

The effects of four food additives, namely sodium nitrite (NaNO2), sodium nitrate (NaNO3), potassium nitrite (KNO2), and potassium nitrate (KNO3), on animal development were evaluated by using Drosophila melanogster, a model organism. Adult male and female flies were allowed to breed in culture medium, each containing one of 4 concentrations, i.e.10, 20, 30 or 40 mM of the above mentioned salts. The concentration of 40 mM, NaNO2 and KNO2 completely arrested the development of the flies. Of the different concentrations of the four salts tested, exposure of flies to 30 mM NaNO2 exhibited only significant delays in the initial appearances of third instar larvae, pupae and young adults, along with huge reduction in the number of pupae and young adults compared to controls. Rearrangements like inversions, deletion looping, regional shrinking, as well as highly enlarged puffing, etc. were also observed in the polytene chromosomes of the third instar larvae exposed to 30 mM NaNO2. Developmental outcomes of the flies exposed to varying concentrations of NaNO3 and KNO3 did not differ significantly from the controls. Owing to the extensive genetic homology between Drosophila and human and the successful uses of this fly as models in developmental and toxicological studies, we speculate that the experimental results exhibited by this organism in our study strongly advocate for abstaining from the dietary use of NaNO2 and KNO2 during human pregnancies to avoid possible negative developmental outcomes.

A FACS Based Case Study on Two HbE-ß Thalassaemia Members of a Family, Having Similar Mutational Background.

In this report we have tried to explain the reasons behind the difference in the pattern of transfusion requirement between two members of a family with similar ß-globin mutation. The father and younger son both are HbE-ß, but the father never had transfusion, whereas the younger son takes transfusion monthly. Mother and the elder son are HbEE without any history of transfusion. ß-globin mutations of all family members were determined by ARMS-PCR. These were reconfirmed by direct sequencing of ß-globin gene. Father and younger son were found to be Cod 26 (G-A)/IVS 1-5 (G-C), whereas mother and elder son were found to be Cod 26 (G-A)/Cod 26 (G-A). XmnI sequencing also revealed that all members of the family were CC. Then, flow cytometry study of red blood cells (RBCs) was performed to measure the oxidative stress of the RBCs. This study was also done on the light and dense fractions of the RBC population of the father and younger son. It was seen that the younger son suffers severe oxidative stress, which can be explained by his higher transfusion requirement. From our work, we have established the importance of taking oxidative stress of RBCs into consideration to explain the clinical manifestation and progression of haemoglobin related diseases like thalassaemia.

A comprehensive screening program for ß-thalassemia and other hemoglobinopathies in the Hooghly District of West Bengal, India, dealing with 21 137 cases.

We here present a report of population screening programs (January 2012-December 2015) conducted by the Thalassemia Control Unit, Imambara Sadar Hospital, Chinsurah, Hooghly in the Hooghly District of West Bengal, India for prevention of thalassemia. We screened ß-thalassemia (ß-thal) heterozygotes and homozygotes, and Hb E (HBB: c.79G > A)-ß-thal compound heterozygotes. Among 21 137 cases, we found 1968 heterozygotes and 192 homozygotes or compound heterozygotes. Results were evaluated with standard hematological analyses including red cell indices, hemoglobin (Hb) typing and quantification. The participants of the screening program were divided into six groups (children, pre-marriage cases, post-marital cases, family members of affected individuals, family members of carriers and pregnant women). While considering the average frequency of carriers, many reports recorded both related individuals (family members of trait and affected individuals) as well as unrelated individuals such as school children and pregnant women. These would have to be considered separately and only the unrelated individuals taken to estimate carrier frequencies in this article that would give more realistic data on carrier frequency of unrelated individuals.

Population Screening and Prevention Strategies for Thalassemias and other Hemoglobinopathies of Eastern India: Experience of 18,166 cases.

We evaluated population screening programs (1999-2011), conducted by the Thalassaemia Foundation, Kolkata, India, for the first time in Eastern India in different districts of West Bengal, for prevention of thalassemia comprising screening of heterozygotes and ß-thalassemia intermedia (ß-TI) cases [ß(+), ß(++), ß(0)/ß(+), ß(E)/ß(E) (codon 26 or HBB: c.79G > A), Hb-E-ß-thalassemia (Hb E-ß-thal)]. Among 18,166 cases, we found 2092 heterozygotes and 2245 ß-TI individuals (who had no information about their disorders). Results were evaluated with standard hematological analyses including erythrocyte indices, hemoglobin (Hb) typing and quantification. Participants were divided into five groups (children, pre-marriage cases, pre-pregnancy cases, affected family members, pregnant women). The objectives of this evaluation were to fix cut-off values of red blood cells (RBCs), mean corpuscular volume (MCV), mean corpuscular Hb (MCH), red blood cell distribution width (RDW) and Hb A2, as the standard World Health Organization (WHO) guidelines were not strictly followed in mass-scale screening programs. We have observed many dilemmas in considering the status of the thalassemia subject, due to presence of some other clinical conditions such as iron deficiency anemia, α-thalassemia (α-thal), δ-thalassemia (δ-thal), clinically silent Hb variants, and some cases of non hemoglobinopathies (such as pregnancy) along with thalassemia. The MCV values varied greatly in different conditions of hemoglobinopathies, whereas MCH provided a more stable measurement. We found an MCH value of <27.0 pg is a suitable cut-off point for screening in this population. Participants with an MCH of <27.0 pg should be investigated further to confirm or exclude a diagnosis of ß-thal trait.

Mutation Spectrum of ß-Thalassemia and Other Hemoglobinopathies in Chittagong, Southeast Bangladesh.

Thalassemia is one of the most common autosomal recessive blood disorders in the world. It shows a variety of clinical expression, starting from asymptomatic to severe blood transfusion dependence. More than 500 alleles have been characterized in or around the ß-globin region. Moreover, most geographical regions have their own characteristics, frequency and availability of these alleles, predominantly circulating within the communities present in that particular region. In this study, we explored the spectrum of ß-thalassemia (ß-thal) alleles present in Chittagong, Southeast Bangladesh. This study comprises ß-thal and Hb E (HBB: c.79 G > A) patients from in and around the area of Chittagong. Not only exploring the complete ß-globin mutation spectrum of the area, but we also tried to look at the origin of the mutated alleles. The ß-thal mutations of Bangladesh show a relatively wide spectrum of alleles, which further demonstrates the heterogeneity of the disease in this country. Although our study showed that the majority of the mutations have their origin in neighboring countries such as India, countries of Southeast Asia, Pakistan, etc., some unusual alleles do not originate in neighboring countries and put a little more diversity in the overall spectrum of ß-thal-specific alleles. Overall, this study demonstrates the mutation spectrum related to ß-thal in Chittagong, Southeast Bangladesh.

Mutational spectrum of thalassemias and other hemoglobinopathies in West Bengal, Eastern India.

Thalassemia, an autosomal recessive blood disease, shows a variety of clinical expression in terms of asymptomatic to severe blood transfusion dependence. More than 500 alleles have been characterized in or around the ß-globin region. Most of the geographical regions have their own characteristic alleles that predominantly circulate within the communities present in that particular region. In this article, we try to throw some light to explore the spectrum of ß-thalassemia (ß-thal) alleles present in West Bengal, the eastern part of India. This study comprises thalassemia carriers and diseased persons from different districts of West Bengal. We not only explored the complete mutational spectrum of this state but we also tried to fix the critical range of the values of different hematological parameters [Hb A2, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH)] for the heterozygotes or carriers of ß-thal with the same mutational background. At the same time, we also tried to evaluate the maximum weighted frequency of these parameters for the heterozygotes or carriers of ß-thal with the same mutational background, so that by observing these cut-off values of standard hematological parameters, we were able to predict the carrier or diseased status for mass scale screening and also try to correlate the values of these parameters with different combinations of ß-thal mutation-specific alleles that can be more informative in mass scale (carrier) screening.

Clinical presentation of two ß-thalassemic Indian patients with 1p36 deletion syndrome: Case report.

Here, we present two thalassemic patients (one male and one female), having unusual clinical phenotypes. Both had mental retardation in which one was associated with microcephaly and other had congenital cataract. They were referred to our institute for clinical evaluation and cytogenetic testing. Both patients were tested for presence of abnormal hemoglobin by high performance liquid chromatography and found to be thalassemic. Their ß-globin mutation was also determined by amplification refractory mutation system-polymerase chain reaction. The male patient was found to have intervening sequence 1-5 (G-C)/+, indicating ß-thalassemia trait and the female was found to have Cod 26 (G-A)/IVS 1-5 (G-C), indicating hemoglobin E-ß thalassemia. Their cytogenetic analysis of blood lymphocytes were studied with high-resolution GTG-banding analysis by using chromosome profiling (Cyto-vision software 3.6) on their chromosomes. Results revealed 46,XY,del(1)(p36.21) in the male and 46,XX,del(1)(p36.3) in the female. Their genotype variation showed (based on genome browser) significant gene loss which probably leads to marked phenotype variation. We believe, thalassemia with mental retardation associated with microcephaly and congenital cataract, both having loss in chromosome 1, p36 position, is reported probably first time from India. This report will definitely enlighten all concerns and add to the information in growing literature.