Effects of air exposure, time, temperature, and humidity on different serum electrolytes

Background: Pre-analytical, analytical, or post analytical variations can induce, change, or alter the tests results. Laboratory errors lead to unnecessary delays in test report and also increased costs by repeat samples which have become a pain to the patients. Aims and Objectives: The aims of this study were to determine alterations in the concentration of serum sodium (Na+), potassium (K+), and ionized calcium (Ca++) concentration with reference to air exposure, time, temperature, and humidity. Materials and Methods: Fifty samples as case and 50 samples as control were included from a normal healthy population in this study. After getting the samples, first readings were taken for case samples and were uncapped and the remaining samples were set aside capped at 24°C, 20% humidity for half an hour and followed by second reading which was taken. Results: Variation in the mean serum sodium between groups is 0.06 mEq/L (0.04%) and 0.08 mEq/L (0.07%) which is very negligible and insignificant (P > 0.05). The mean level of serum K+ in cases is 4.35 mEq/L and in controls is 4.27 mEq/L. After half an hour, the mean level of serum K+ in cases is 4.51 mEq/L and, in controls, is 4.29 mEq/L. Hence, the variation in results in cases is 0.16 mEq/L (3.68%) and in controls is 0.02 mEq/L (0.47%) which is highly significant (P < 0.05). The mean level of serum Ca++ in cases is 1.15 mmol/L and in controls is 1.17 mmol/L. After half an hour, the mean level of serum Ca++ in cases is 1.09 mmol/L and in controls is 1.16 mmol/L. Hence, the variation in results in cases is 0.06 mmol/L (5.22%) and in controls is 0.01 mmol/L (0.85%) which is highly significant (P < 0.05). Conclusion: Air exposure significantly alters the serum K+ and Ca++ level, but the alteration in serum Na+ level is not significant.

Characterization and evaluation of apoptotic potential of double gene construct pVIVO.VP3.NS1.

Viral gene oncotherapy, targeted killing of cancer cells by viral genes, is an emerging non-infectious therapeutic cancer treatment modality. Chemo and radiotherapy in cancer treatment is limited due to their genotoxic side effects on healthy cells and need of functional p53, which is mutated in most of the cancers. VP3 (apoptin) of chicken infectious anaemia (CIA) and NS1 (Non structural protein 1) of Canine Parvovirus-2 (CPV-2) have been proven to have oncolytic potential in our laboratory. To evaluate oncolytic potential of VP3 and NS1 together these genes needed to be cloned in a bicistronic vector. In this study, both these genes were cloned and characterized for expression of their gene products and its apoptotic potential. The expression of VP3 and NS1 was studied by confocal microscopy and flowcytometry. Expression of VP3 and NS1 in pVIVO.VP3.NS1 transfected HeLa cells in comparison to mock transfected cells indicated that the double gene construct expresses both the products. This was further confirmed by flowcytometry where there was increase in cells expressing VP3 and NS1 in pVIVO.VP3.NS1 transfected group in comparison with the mock control group. The apoptotic inducing potential of this characterized pVIVO.VP3.NS1 was evaluated in human cervical cancer cell line (HeLa) by DNA fragmentation assay, TUNEL assay and Hoechst staning. This double construct was observed to induce apoptosis in HeLa cells.

A clinical study on amoebic liver abscess.

Background: Amoebic liver abscess presents with severe pain and high grade fever and if not diagnosed and treated promptly, may lead to complications and mortality. Aim and objectives: The objective of the present study was to estimate the incidence, need for aspiration and prognosis. The diagnosis was based on clinical features, positive Elisa test, ultrasonography, aspiration of anchovy sauce from the liver lesion, isolation of E. Histolytica (cyst/trophozoite) from the stool of the patient. Result: We had 65 cases in the study. There were 52 males & 13 females with a ratio of 4:1. Solitary abscess was found in 48(73.8%) patients which are located as follows; right lobe(43), left lobe(2) and in both lobe(3). 9% were aspirated at presentation due to their size or position. Only 4 (2%) were aspirated at first follow-up on third day due to non resolution of pain or fever or increase in size. All the patients are responded to standard treatment of metronidazole. Amoebic liver abscess is a common diagnosis in our setup. Conclusion: Clinical background and sonogram give a reasonable suggestion about amoebic etiology. If initial aspiration is not indicated due to size larger than10 cm or proximity to surface, conservative treatment with oral or intravenous metronidazole is successful.

Investigation of Nutrient Content, Phytochemical Content, Antioxidant Activity and Antibacterial Activity of Inedible Portion of Pomegranate (Punica granatum L.).

Pomegranate is a plant with high potential to have antimicrobial and antioxidant compounds. In the present study inedible parts (peel and seed) of pomegranate, variety “Bhagawa” were examined for the presence of phytochemicals, phenolic compounds, flavonoids, anthocyanin, antioxidant and anti-microbial compounds. Further, nutrition value of the samples was also examined. Results of GC-MS analysis carried out using four different solvents showed 12 and 25 fractions for peel and seed extracts respectively. Both peel and seed were with high amount of Anthocyanin, Phenolic compounds and Flavonoids. Both extracts gave 100% free radicals reduction in DPPH assay showing high antioxidant activity. Further, both extracts showed antibacterial activity against eight tested food borne pathogens. Antibacterial activity shown by peel extracts was higher than that of seed extract. Both peel and seed extracts showed highest and lowest antibacterial activity against Staphylococcus aureus and Listeria monocyatogenes respectively. Both extract showed same antibacterial activity which was shown by Kanamycin for Shigella fexneri. According to the present study, pomegranate peel and seed can be used to extract antibacterial, antioxidant compounds and flavonoids for the use in food industry.

Acute Promyelocytic Leukemia in a 17yr Male presenting as Intracerebral Haemorrhage.

Acute promyelocytic leukemia (APML), once highly fatal, has emerged as the most curable subtype of acute myeloid leukemia in adults. Early mortality most often is due to a severe catastrophic bleeding, often intracerebral in location. Here we report a 17year male patient presented with status epilepticus having high leukocyte count (3,28,000/mm3) and low platelet count(34,000/mm3).Peripheral blood & bone marrow was showing good no. of atypical promyelocytes. CT scan of brain revealed an intracerebral haemorrhage with laboratory profile of prolonged prothrombin time and activated partial thromboplastin time, the presence of Ddimer and decreased fibrinogen concentration. The patient was diagnosed as Acute Promyelocytic Leukemia with Intracerebral haemorrhage. The patient died on the same day. APML is the notorious subtype of acute myeloid leukemia which causes fatal intracranial haemorrhage which has high mortality and morbidity. Clinically significant coagulopathy is present in 70%– 80% of APML patients at the time of diagnosis. Early detection and aggressive correction of coagulopathy may prevent the catastrophic event. Prompt image study for locations and types of ICH can predict outcomes.

Cloning and expression analysis of multiple proteins encoding P gene of Newcastle disease virus.

Viral gene oncotherapy is emerging as a biotherapeutic cancer treatment modality based on targeted killing of cancer cells by viral genes. Newcastle disease virus (NDV) has the property to cause selective oncolysis of tumor cells sparing normal cells. NDV has a single stranded negative sense RNA genome, which is 15,186 nucleotide long and consists of six genes, which codes for eight proteins. NDV like other paramyxoviruses has the ability to generate multiple proteins from the P gene. P protein is encoded by an unedited transcript of the P gene, whereas the V and W protein are the results of RNA editing event in which one and two G residues are inserted at a conserved editing site within the P gene mRNA resulting in V and W transcripts, respectively. Although NDV is known to cause oncolysis by triggering apoptosis, the role of different viral proteins in selective oncolysis is still unclear. P gene edited products are known for its anti-apoptotic property in homologous host. In the present study, NDV P gene and its RNA edited products were amplified, cloned, sequenced and in vitro expression was done in HeLa cells. Further constructs were assayed for their apoptosis inducing ability in HeLa cells. Preliminary study suggested that P, V and W proteins are not apoptotic to HeLa cells.

Oncolytic viruses & their specific targeting to tumour cells.

Cancer is one of the major causes of death worldwide. In spite of achieving significant successes in medical sciences in the past few decades, the number of deaths due to cancer remains unchecked. The conventional chemotherapy and radiotherapy have limited therapeutic index and a plethora of treatment related side effects. This situation has provided an impetus for search of novel therapeutic strategies that can selectively destroy the tumour cells, leaving the normal cells unharmed. Viral oncotherapy is such a promising treatment modality that offers unique opportunity for tumour targeting. Numerous viruses with inherent anti-cancer activity have been identified and are in different phases of clinical trials. In the era of modern biotechnology and with better understanding of cancer biology and virology, it has become feasible to engineer the oncolytic viruses (OVs) to increase their tumour selectivity and enhance their oncolytic activity. In this review, the mechanisms by which oncolytic viruses kill the tumour cells have been discussed as also the development made in virotherapy for cancer treatment with emphasis on their tumour specific targeting.

Characterization and in vitro expression of non-structural 1 protein of canine parvovirus (CPV-2) in mammalian cell line.

Parvoviruses are small, 260-Å-diameter, icosahedral, non-enveloped, single-stranded DNA viruses with a genome of approximately 5 kb. Non structural protein, (NS-1) is especially relevant, being both essential for virus replication and the main factor responsible for virus pathogenicity and cytotoxicity. This protein has also been reported to possess the property of killing of transformed cells. The present study was carried out to clone, characterize and express the NS-1 gene of canine parvovirus. NS-1 complete CDS 2020bp was amplified, cloned into eukaryotic expression vector pcDNA 3.1(+), sequenced and characterized by in vitro expression analysis. Functional activity of recombinant construct, pcDNA.cpv.NS-1, was evaluated by RT-PCR and flow cytometry for the expression of NS-1 specific mRNA and NS-1 protein, respectively, in transfected HeLa cells. This recombinant plasmid may serve as an important tool to evaluate the apoptotic potential of NS-1 protein of canine parvovirus in cultured HeLa cells.

Development and in vitro characterization of a bivalent DNA containing HN and F genes of velogenic Newcastle disease virus.

Newcastle disease (ND) is highly contagious, economically important viral disease affecting most of avian species worldwide. Newcastle disease virus (NDV) has single stranded negative sense RNA genome which encodes for six structural and two non-structural proteins. Envelope glycoproteins i.e. hemagglutinin-neuraminidase (HN) and the fusion (F), elicit protective immune response. In this study, HN and F genes of velogenic (virulent) strain were amplified and cloned at multiple cloning sites A and B, respectively into pIRES bicistronic vector for use as bivalent DNA vaccine against ND. The recombinant plasmid was characterized for its orientation by restriction enzyme digestion and PCR. Expression of HN and F genes was assessed in transfected Vero cells at RNA level using RT-PCR in total RNA as well as protein level using IFAT, IPT and western blot using NDV specific antiserum. All these experiments confirmed that HN and F genes cloned in recombinant pIRES.nd.hn.f are functionally active. The recombinant construct is being evaluated as DNA vaccine against ND.

Cloning and expression of chicken granulocyte-macrophage colony stimulating factor (GMCSF) gene.

Granulocyte-macrophage colony stimulating factor (GMCSF), a multifunctional cytokine can enhance immune responses when administered along with DNA vaccine. Aim of the present study was to clone and express the chicken GMCSF cytokine for use as ‘genetic adjuvant’. Chicken GMCSF gene 435bp was amplified using specific primers in which restriction sites of BamHI and HindIII were at forward and reverse primers respectively. The PCR product was cloned into eukaryotic expression vector pcDNA 3.1(+) and clones were confirmed by restriction digestion and nucleotide sequencing. Functional activity of recombinant GMCSF was checked by expression of GMCSF specific mRNA in transfected Vero cells by RT-PCR of total RNA isolated from transfected Vero cells. The recombinant plasmid can be used as genetic adjuvant in chicken.

Incidence of leprosy in school-children and their family members in Berhampur.

A school survey, followed by a contact survey, was carried out in Berhampur, a city in southern Orissa. In a study of 8,870 school-children, leprosy was detected in 15, giving a prevalence rate of 16.91 per 10,000 with a male:female ratio of 8:7. Of these, 14 (93.99%) had paucibacillary leprosy. More cases [11 (73.33%)] were seen in the age-group of 10-15 years. Exposed parts, such as lower limbs, upper limbs and head and neck in that order, were the sites of predilection, accounting for 85.71% of total lesions. Nerve involvement was found in 2 (13.33%) girls with deformity (ulnar claw) in one of them (6.66%). BCG scar was present in 11 (73.33%) cases. Among the vaccinated cases, tuberculoid type was the most common, followed by indeterminate, pure neuritic and borderline, in that order. A contact survey detected 2 multibacillary cases in two families (13.33%). In each case, the father was the index source. The study revealed that a maximum number of students, 8 (53.3%), belonged to the middle socioeconomic class. Of the 15 affected, 60% were undernourished and the rest well nourished. No other systemic disease was found clinically associated with leprosy.