Anemia Awareness Program in a Suburban Middle School: A Pilot Study Promoting Students as Public Health Advocates in Their Community.

INTRODUCTION: Almost a quarter of the people on earth are anemic, and most of them reside in regions of sub-Saharan Africa and South Asia. Anemia in children is linked with impaired cognitive and motor development and affects the future earning capacity. The most common cause of anemia is iron deficiency. The Indian Government has initiated multiple programs for the eradication of anemia. The prevalence of anemia has not decreased despite the improvements in the country's economy. It increased from 58.7% in 2015-16 to 67.1% in 2019-21 in children and from 50.4% in 2015-16 to 52.2% in 2019-21 in pregnant women. Maternal education, socioeconomic status, and number of children in the family are some factors that influence the prevalence of anemia. As these factors cannot be improved in a short time, we aimed to increase awareness about this issue by targeting school students from rural/semi-urban backgrounds. METHODS: This pilot study aimed at promoting school students as public health advocates in their community. Anemia Awareness Program was conducted in a local middle school in the suburban area, which was attended by 153 class eight students (72 female). Pre- and post-test questionnaires comprising 20 multiple-choice/true-false type questions were used. Pre- and post-test scores were obtained. The second part of the study was the identification of students with anemia. Blood hemoglobin levels of 127 students (58 female) were measured from venous blood samples. The students were also asked to inform their friends/relatives about anemia and to send people with symptoms of anemia to the free two-day Anemia Awareness Camp organized by the Medical College Hospital for check-ups. RESULTS: The mean post-test score (15.68/20) was much higher than the pretest score (2.99/20). Thirty-eight (25 female) out of 127 students had mild/moderate microcytic hypochromic anemia, suggesting iron deficiency. Thirty-two persons visited the free health camp to receive information from the students, of whom four had normal hemoglobin levels. CONCLUSION: This pilot study showed that physician-conducted anemia awareness programs are relatively low-cost methods to spread information among the general population in India.

Immunological and safety profile of bacteriophage therapy: A pre-clinical study.

AIMS: Numerous pre-clinical and clinical studies have recently demonstrated the significant role of phage therapy in treating multidrug-resistant bacterial infections. However, only a few researchers have focused on monitoring the phage-mediated adverse reactions during phage therapy. The present study aimed to demonstrated the oral acute and sub-acute toxicity of bacteriophages (Klebsiella pneumoniae XDR strain) in Charles Foster rats with special reference to immunological response and adverse effects. METHODS AND RESULTS: Bacteriophages were orally administered in dosages of 1010  PFU/ml and a 1015  PFU/ml to Charles Foster rats as a single dose (in acute toxicity study) and daily dosage for 28 days (in sub-acute toxicity study). One millilitre suspension of bacteriophages was administered through the oral gavage feeding tube. No adverse effect was observed in any of the experimental as well as in the control animals. Furthermore, an insignificant change in food and water intake and body weight was observed throughout the study period compared with the control group rats. On the 28th day of phage administration, blood was collected to estimate haematological, biochemical and cytokines parameters. The data suggested no difference in the haematological, biochemical and cytokine profiles compared to the control group. No significant change in any of the treatment groups could be observed on the gross and histopathological examinations. The cytokines estimated, interleukin-1 beta (IL-1ß), IL-4, IL-6 and IFN-gamma, were found within the normal range during the experiment. CONCLUSIONS: The results concluded that no adverse effect, including the severe detrimental impact on oral administration of high (1010 PFU/ml) and very high dose (1015  PFU/ml) of the bacteriophages cocktail. SIGNIFICANCE AND IMPACT OF STUDY: The high and long-term oral administration of bacteriophages did not induce noticeable immunological response as well.

Vitality of Proteinase K in rRTPCR Detection of SARS-CoV2 Bypassing RNA Extraction.

This study aimed to detect the SARS-COV2 viral component directly from inoculated VTM without RNA extraction. Inoculated VTMs of already tested 50 positive and 50 negative samples were divided into three groups. Group I was treated with Proteinase K (PK) followed by 3-step-heat treatment at different temperatures (25°C, 60°C, and 98°C) and stored at 4°C. Group II was directly subjected to 3-step-heat treatment without PK exposure and stored at 4°C. And group III was set-up as standard group; it was processed using Qiagen's column based QIAamp Nucleic Acid kit and the obtained nucleic acids were stored at 4°C. These stored samples were used as a template to execute real-time polymerase chain reaction, and results were noted. Group I demonstrated 96% and 88% sensitivity for N and ORF1ab genes respectively, whereas group II demonstrated 78% and 60% when compared to the results of standard group III. Overall group I showed better results than group II when compared to group III. Thus, in situations where gold-standard reagents are not available, PK exposure and heat treatment can be employed to carry out molecular detection of SARS-CoV2 viral component.

Assessment of Successful qRT-PCR of SARS-CoV-2 Assay in Pool Screening Using Isopropyl Alcohol Purification Step in RNA Extraction.

The study is aimed at establishing the optimal parameters for RNA purification of pooled specimens, in SARS-CoV-2 assay. This research work evaluates the difference of extracted RNA purity of pooled samples with and without treatment with isopropyl alcohol and its effect on real-time RT-PCR. As per the protocol of the Indian Council of Medical Research (ICMR), 5 sample pools were analysed using qRT-PCR. A total of 100 pooled samples were selected for the study by mixing 50 µL of one COVID-19 positive nasopharyngeal/oropharyngeal (NP/OP) specimen and 50 µL each of 4 known negative specimens. Pool RNA was extracted using the column-based method, and 1 set of pooled extracted RNA was tested as such, while RNA of the second set was treated additionally with chilled isopropyl alcohol (modified protocol). Further, the purity of extracted RNA in both the groups was checked using Microvolume Spectrophotometers (Nanodrop) followed by RT-PCR targeting E-gene and RNaseP target. The results showed that the purity index of extracted RNA of untreated pooled specimens was inferior to isopropyl alcohol-treated templates, which was observed to be 85% sensitivity and 100% specificity. The average Cq (E gene) in the unpurified and purified pool RNA group was 34.66 and 31.48, respectively. The nanodrop data suggested that purified RNA concentration was significantly increased with an average value of 24.73 ± 1.49 ng/uL, which might be the reason for high sensitivity and specificity. Thus, this group testing of SARS-CoV-2 cases using pools of 5 individual samples would be the best alternative for saving molecular reagents, personnel time, and can increase the overall testing capacity. However, purity of RNA is one of the important determinants to procure unfailing results, thus, this additional purification step must be included in the protocol after RNA has been extracted using commercially available kit before performing qRT-PCR.

Study on the Effect of Oral Administration of Bacteriophages in Charles Foster Rats With Special Reference to Immunological and Adverse Effects.

Numerous pre-clinical and clinical studies have recently demonstrated the significant role of phage therapy in treating multidrug-resistant bacterial infections. However, only a few researchers have focused on monitoring the phage-mediated adverse reactions during phage therapy. Besides adverse reactions, immunological response after short- and long-term oral administration of bacteriophages is also lacking. In this study, we administered the bacteriophages orally against Klebsiella pneumoniae XDR strain in dosages of 1015 PFU/ml and a 1020 PFU/ml (still higher) to Charles Foster rats as a single dose (in acute toxicity study) and daily dosage for 28 days (in sub-acute toxicity study). One milliliter suspension of bacteriophages was administered through the oral gavage feeding tube. No adverse effect was observed in any of the experimental as well as in the control animals.Further, an insignificant change in food and water intake and body weight was observed throughout the study period compared with the control group rats. On the 28th day of phage administration, blood was collected to estimate hematological, biochemical, and cytokines parameters. The data suggested no difference in the hematological, biochemical, and cytokine profile compared to the control group. No significant change in any of the treatment groups could be observed on the gross and histopathological examinations. The cytokines estimated, interleukin-1 beta (IL-1ß), IL-4, IL-6, and INF-gamma, were found within the normal range during the experiment. The results suggested no adverse effect, including the severe detrimental impact on oral administration of high (1015 PFU/ml) and very high dose (1020 PFU/ml) of the bacteriophages cocktail. The high and long-term oral administration of bacteriophages did not induce noticeable immunological response as well.

Association of single nucleotide polymorphisms in vascular endothelial growth factor gene with bladder cancer risk.

Vascular endothelial growth factor (VEGF) is an endothelial cell-specific potent mitogen involved in a number of pathologic processes, including angiogenesis, tumor growth, and metastasis. Polymorphisms of VEGF gene have been associated with susceptibility to several cancers. VEGF gene susceptibility to bladder cancer (BC), however, still remains controversial. We analyzed association of the -2578C/A, -7C/T, -2549I/D, and -1001G/C polymorphism of the VEGF gene with bladder cancer (BC) risk and interaction of these polymorphisms with lifestyle and demographic factors. We genotyped -2578C/A, -7C/T, -2549I/D and -1001G/C by PCR-restriction fragment length polymorphism and amplification refractory mutation specific method to evaluate risk in 200 BC patients and 250 healthy controls from North Indian population. Significant association for BC risk in heterozygous CA genotype (1.69-fold) in VEGF-2578C/A and heterozygous genotype of VEGF-1001G/C (p = 0.032) was observed. Interestingly, VEGF-2549I/D genotype showed reduced risk for BC. The gene-gene combination analysis revealed DD-GG with reduced risk (p = 0.018) of VEGF-2549I/D and VEGF-1001G/C, and combination CA-GG of VEGF-2578C/A and VEGF-1001G/C demonstrated 1.75-fold risk for BC. Our findings suggested that polymorphism -2578C/A and -1001G/C in the promoter of VEGF gene may play a significant role in mediating the bladder cancer risk, whereas VEGF-2549I/D genotype appears to be protective for BC.

Exocytosis, dependent on Ca2+ release from Ca2+ stores, is regulated by Ca2+ microdomains.

The relationship between the cellular Ca2+ signal and secretory vesicle fusion (exocytosis) is a key determinant of the regulation of the kinetics and magnitude of the secretory response. Here, we have investigated secretion in cells where the exocytic response is controlled by Ca2+ release from intracellular Ca2+ stores. Using live-cell two-photon microscopy that simultaneously records Ca2+signals and exocytic responses, we provide evidence that secretion is controlled by changes in Ca2+ concentration [Ca2+] in relatively large-volume microdomains. Our evidence includes: (1) long latencies (>2 seconds) between the rise in [Ca2+] and exocytosis, (2) observation of exocytosis all along the lumen and not clustered around Ca2+ release hot-spots, (3) high affinity (Kd=1.75 microM) Ca2+dependence of exocytosis, (4) significant reduction in exocytosis in the presence of cytosolic EGTA, (5) spatial exclusion of secretory granules from the cell membrane by the endoplasmic reticulum, and (6) inability of local Ca2+ responses to trigger exocytosis. These results strongly indicate that the control of exocytosis, triggered by Ca2+ release from stores, is through the regulation of cytosolic[Ca2+] within a microdomain.

Pancreatic acinar cell: new insights into the control of secretion.

Pancreatic acinar cells secrete fluid and digestive enzymes. Both types of secretion are activated by a rise in intracellular calcium but how the stimulus-secretion cascade actually regulates secretory output is not well understood. It has long been known that the calcium response of acinar cells to physiological stimulation is complex. Dependent on the type and concentration of agonist, it consists of either local or global calcium increases as well as spreading waves of calcium across the cell. In the past it has been speculated that these different calcium signals drive different secretory responses. Now, recent employment of two-photon microscopy has enabled the simultaneous recording of both enzyme secretion and calcium signals and is beginning to resolve this issue. The data shows that local calcium responses exclusively drive fluid secretion. Where-as, global calcium responses drive both fluid and enzyme secretion. This differential control of secretory output is likely central to controlling the physiological responses of pancreatic acinar cells.

Local dynamic changes in confined extracellular environments within organs.

1. Herein we review past work that has studied the composition of luminal fluid in organs, with a focus on measures of calcium and pH in the exocrine glands. This luminal environment is 'external' to the mammalian body and is not subject to the usual mechanisms of homeostatic control. Instead, it is controlled by the behaviour of the cells that line the lumen. 2. We discuss the likely possibility that rapid and local changes in calcium and pH occur within microdomains in the lumen. Further, we present preliminary evidence, using live cell imaging of intact pancreatic fragments, that supports the idea that pH changes do occur. Our evidence indicates that exocytosis of secretory granules in pancreatic acinar cells leads to a loss of protons from the granule and a subsequent local acidification of the lumen. 3. These changes in luminal composition are placed in the context of diseases of the pancreas, such as cystic fibrosis and pancreatitis, both of which are known to result in perturbations of luminal fluid composition.

Removal of nickel from aqueous solutions by sawdust.

The main parameters influencing Nickel (II) metal sorption on maple sawdust were: initial metal ion concentration, amount of adsorbent, and pH value of solution. The maximum percent metal removal was attained after about 1h. The greatest increase in the rate of adsorption of metal ions on sawdust was observed for pH changes from 2 to 5. An empirical relationship has been obtained to predict the percentage Nickel (II) removal at any time for known values of sorbent and initial sorbate concentration. The experimental results provided evidence for chelation ion exchange as the major adsorption mechanisms for binding metal ions to the sawdust. The adsorbent can be effectively regenerated using 0.1 M strong acid and reused.

Rapid detection of sewage sample polioviruses by integrated cell culture polymerase chain reaction.

The authors' objective in this study was to introduce and evaluate integrated cell culture polymerase chain reaction (ICC-PCR) as a technique for the rapid screening of poliovirus in sewage samples. Researchers are in the last stage of poliomyelitis eradication; however, in a densely populated country such as India, time is the prime factor in the identification of poliovirus circulation and transmission because this virus follows the fecal-oral route for transmission and is excreted in nature. The authors used ICC-PCR to detect poliovirus in sewage samples, and they compared this nonconventional method with conventional cell culture methods to determine sensitivity, accuracy, and the time from sample collection to final results. The ICC-PCR method provided results within 4-5 days of sewage-sample collection; in contrast, the conventional method takes more than 18 days to provide such results. The ICC-PCR method proved to be sensitive, reproducible, and accurate, as well as rapid in its screening of sewage samples for poliovirus. This diagnostic tool may indeed prove quite useful in polio eradication.

Adsorption of chromium from aqueous solutions by maple sawdust.

This paper presents the data for the effect of adsorbent dose, initial sorbate concentration, contact time, and pH on the adsorption of chromium(VI) on maple sawdust. Batch adsorption studies have been carried out. An empirical relationship has been obtained to predict the percentage chromium(VI) removal at any time for known values of sorbent and initial sorbate concentration. Under observed test conditions, the equilibrium adsorption data fits the linear Langmuir and Freundlich isotherms. The experimental result inferred that chelation ion exchange is one of the major adsorption mechanisms for binding metal ions to the maple sawdust.

The role of sawdust in the removal of unwanted materials from water.

Sawdust, a relatively abundant and inexpensive material is currently being investigated as an adsorbent to remove contaminants from water. Chemical substances including dyes, oil, toxic salts and heavy metals can be removed very effectively with the organic material. This article presents a brief review on the role of sawdust in the removal of contaminants. Studies on the adsorption of various pollutants by different sawdust materials are reviewed and the adsorption mechanism, influencing factors, favorable conditions, etc. discussed in this paper. Some valuable guidelines can be drawn for either scientific research or industrial design.