STABILITY INDICATING METHOD DEVELOPMENT AND VALIDATION FOR ESTIMATION OF DORIPENEM IN BULK AND IN SOLID DOSAGE FORM BY RP-HPLC

A robust and reliable high performance liquid chromatographic (HPLC) approach was developed and validated for the analysis of Doripenem in pharmaceutical dosage form. The method is characterised by its simplicity, selectivity, precision, and capacity to accurately determine the stability of Doripenem. The experimental setup included the use of a Hypersil BDS-C18 column (250 X 4.6 mm ID, 5 µm) as the stationary phase in a chromatographic system. The mobile phase consisted of a combination of methanol and potassium dihydrogen orthophosphate with a pH of 6.7, in a ratio of 20:80. The flow rate of the mobile phase was set at 1 ml/min. The detection of the eluents occurred at a wavelength of 290 nm. The observed retention time for Doripenem was 5.56 minutes. Doripenem underwent acid and alkali hydrolysis, oxidation, photochemical degradation, and heat degradation. The results obtained from the linear regression analysis of the calibration plot demonstrated a strong linear connection within the concentration range of 70 – 130 µg/ml, as shown by a correlation coefficient value of 0.9995. The methodology was assessed to determine its precision, accuracy, ruggedness, and robustness. The medication experiences deterioration when exposed to environments characterised by acidity, alkalinity, photochemical reactions, and thermal stress. The active medicinal component exhibited distinct retention periods for each of its degradation product peaks, indicating successful resolution. The approach's ability to successfully isolate the medication from its degradation products renders it suitable for use as a stability-indicating method

A Clinical Study on Pipaladi Leh in Tamak Shwasa (Bronchial Asthma) in Children

Tamak Shwasa is a term that encompasses many diseases such as shortness of breath as the main symptom. But we can also associate bronchial asthma with Tamak Shwasa because of the remaining four, three of them are incurable and the fourth is Kshudra Shwasa, which is caused by overwork and overeating, it can be cured in a simple way. Early onset is easy to treat, but the chronic condition is difficult to treat. In Ayurveda, the word Shwasa defines its meaning as a disease in itself as well as symptoms and problems of other diseases. The patient has an attack of a disease that threatens his life. Acharya Charaka describes Tamak Shwasa as a Yappya type of illness in which the patient has to rely on medicine for relief. Acharya Charak teaches different good thoughts about Tamak Shwasa. Every Ayurvedic practitioner should have a detailed knowledge of Tamak Shwasa to distinguish between Chikitsa and disease assessment. In Pipalaadi leh therapy, 04 pts. i.e., 09.76% of the patients attained marked improvement, 33 pts. i.e., 80.48% of the patients attained moderate improvement and 04 pts. i.e., 09.76% of the patients attained mild improvement

Effect of Shashtika Shali Pinda Sweda & other Ayurvedic Intervention in Cerebral palsy: A Case Report

Cerebral palsy is a disorder that affects muscle tone, movement and motor skills (the ability to move in a coordinated and purposeful way). Cerebral Palsy cannot be correlated with any single disease or condition mentioned in Ayurveda, as it is a multi-factorial disease. The classical signs and symptoms of CP will fit into criteria of Vata Vyadhi spectrum (Vata predominant disease) like Ekangvata, Sarvangvata, Pakshaghata, Pangu, Kampavata etc and Phakkaroga. Although cerebral palsy cannot be completely cured but treatment will generally enhance a child’s abilities. Agnimandya, Amavastha and Kaphavastha should be considered while planning the line of treatment in CP cases. The selected Ayurvedic treatment modality is highly effective in relieving the signs and symptoms and thus reducing the disability in children with CP. Swedana (sudation) is the therapy that relieves the Stambha (stiffness) of the body, mitigate feeling of Guruta (heaviness) and Sheeta (feeling of cold). Shastika shali panda sweda comes under the category of Saagniseda with Snigda dravya as milk and Shalidhanya. It has Snigdha, Guru, Sthira, Sheeta, Tridoshaghna and Brimhanaguna. In this article, an attempt to treat a child with spastic diplegia using multiple Ayurveda treatment modalities. At the end of one year of treatment, Panchkarma procedures along with internal medication resulted in 15-20% improvement in the overall effect of therapy

Ayurvedic Management of Comorbid State of Autism with Attention Deficit Hyperactivity Disorder w.s.r Unmada

Unmada a term used in Ayurveda, refers to a condition characterized by mental instability, restlessness and abnormal behaviors. It similar to modern day autism spectrum disorder (ASD) and attention deficit hyperactive disorder (ADHD). ASD and ADHD are neuro developmental and neuropsychiatric disorder. These disorders have shared genetic heritability and are both associated with shared impairment in social functioning and executive functioning. However, in the phenotypic presentation of the impairment which characterized ASD and ADHD. The fundamental principles of Ayuvedic management are to restore the balance of the Doshas. In the case of Unmada, particularly when comorbid with autism and ADHD, the focus on pacifying the aggravated Vata dosha. This can be achieved through a combination of dietary modification, lifestyle changed, Balpanchkarma and Marma therapies and Medhya Dravya. Aim and Objective: A case study of Ayurvedic management of comorbid state of Autism and Attention Deficit Hyperactivity Disorder w.s.r Unmaad. Material and Method: A 5-year-old female patient was come to the OPD of Kaumarbhritya Department, Rishikul Campus, UAU, Haridwar. With complain of difficulty in speaking as per development age, poor eye contact, hyperactive in nature with addiction of mobile in the last three year. This condition can be understood as Unmada. Treatment including with some Balpanchkarma and Marma therapies, and Ayurvedic formulations for 2-3 months. Result: There significant improvement in the condition of the patient. Ayurvedic intervention in this case reveals the true potential and efficacy of our Ayurvedic science

Absence of simian virus 40 in human brain tumors from Northern India

Simian virus 40 (SV40), a monkey polyomavirus, was a contaminant of early poliovirus vaccines administered to millions of individuals in the 1950s and early 1960s. SV40 causes brain tumors in laboratory animals, and SV40 DNA sequences have been variably identified in human choroid plexus tumors and ependymomas. We studied the possible association between SV40 and human brain tumors in northern India, where humans have frequent contact with SV40-infected rhesus macaques. DNA from pathologic specimens from 33 ependymomas, 14 choroid plexus tumors and 18 control brain tissues (contused brain, brain metastases) was extracted and analyzed under masked conditions. We used real-time PCR to detect and quantify SV40 (T antigen) and human (GAPDH) DNA sequences. The SV40 PCR assay detected as few as 10 copies of SV40 DNA and had a linear range from 1 × 10<SUP>2</SUP> to 1 × 10<SUP>6</SUP> copies. SV40 DNA was detected in 1 specimen (an ependymoma). However, few SV40 DNA copies were detected in this sample (&lt;10 copies, equivalent to &lt;1 copy/350 cells, based on simultaneous GAPDH quantification), and SV40 was not detected when this sample was retested. Our findings do not support a role for SV40 in choroid plexus tumors or ependymomas from northern India

A Report on Molecular Diagnostic Testing for Inherited Retinal Dystrophies by Targeted Genetic Analyses

AimTo test the utility of targeted sequencing as a method of clinical molecular testing in patients diagnosed with inherited retinal degeneration (IRD).MethodsAfter genetic counseling, peripheral blood was drawn from 188 probands and 36 carriers of IRD. Single gene testing was performed on each patient in a Clinical Laboratory Improvement Amendment (CLIA) certified laboratory. DNA was isolated, and all exons in the gene of interest were analyzed along with 20 base pairs of flanking intronic sequence. Genetic testing was most often performed on ABCA4, CTRP5, ELOV4, BEST1, CRB1, and PRPH2. Pathogenicity of novel sequence changes was predicted by PolyPhen2 and sorting intolerant from tolerant (SIFT).ResultsOf the 225 genetic tests performed, 150 were for recessive IRD, and 75 were for dominant IRD. A positive molecular diagnosis was made in 70 (59%) of probands with recessive IRD and 19 (26%) probands with dominant IRD. Analysis confirmed 12 (34%) of individuals as carriers of familial mutations associated with IRD. Thirty-two novel variants were identified; among these, 17 sequence changes in four genes were predicted to be possibly or probably damaging including: ABCA4 (14), BEST1 (2), PRPH2 (1), and TIMP3 (1).ConclusionsTargeted analysis of clinically suspected genes in 225 subjects resulted in a positive molecular diagnosis in 26% of patients with dominant IRD and 59% of patients with recessive IRD. Novel damaging mutations were identified in four genes. Single gene screening is not an ideal method for diagnostic testing given the phenotypic and genetic heterogeneity among IRD cases. High-throughput sequencing of all genes associated with retinal degeneration may be more efficient for molecular diagnosis

Validation of SYBR green I based closed tube loop mediated isothermal amplification (LAMP) assay and simplified direct-blood-lysis (DBL)-LAMP assay for diagnosis of visceral leishmaniasis (VL).

BackgroundThe World Health Organization has targeted elimination of visceral leishmaniasis (VL) in the Indian subcontinent (ISC) by 2020. Despite distinctive decline seen in the number of VL cases in ISC, there is still a quest for development of a diagnostic test which has the utility for detection of active infection and relapse cases and as a test of cure. The present study validated the sensitivity and specificity of SYBR Green I based closed tube LAMP assay reported by us for diagnosis of VL.MethodologyThe validation study was carried out at two endemic sites in India, located at Rajendra Memorial Research Institute of Medical Sciences (RMRIMS), Patna and Institute of Medical Sciences (IMS), Banaras Hindu University (BHU), Varanasi. Standard operating protocols were provided at the two sites for applying LAMP assay on confirmed VL cases. The diagnostic accuracy of LAMP assay was evaluated by Receiver operator curve (ROC) analysis. Furthermore, a simplified LAMP assay based on direct blood lysis, DBL-LAMP, was developed and verified for its diagnostic accuracy.Principal findingsA total of 267 eligible participants were included in the study which comprised of 179 VL cases and 88 controls. Sensitivity and specificity of the LAMP assay were 98.32% (95% C.I- 95.2-99.7%) and 96.59% (95% C.I.-90.4-99.3%), respectively. ROC curve analysis depicted no significant difference between area under curve (AUCROC) for LAMP assay and rK39 RDT, indicative of LAMP as an excellent diagnostic test. DBL-LAMP assay, performed on 67 VL and 100 control samples, yielded a sensitivity of 93.05% (95% C.I- 84.75-97%) and specificity of 100% (95% C.I.- 96.30-100%).Conclusions/significanceThe validated closed tube LAMP for diagnosis of VL will provide impetus to the ongoing VL elimination programme in ISC. The assay based on direct blood lysis promotes its scope for application in field settings by further reducing time and cost

A Report on Molecular Diagnostic Testing for Inherited Retinal Dystrophies by Targeted Genetic Analyses

Aim: To test the utility of targeted sequencing as a method of clinical molecular testing in patients diagnosed with inherited retinal degeneration (IRD). Methods: After genetic counseling, peripheral blood was drawn from 188 probands and 36 carriers of IRD. Single gene testing was performed on each patient in a Clinical Laboratory Improvement Amendment (CLIA) certified laboratory. DNA was isolated, and all exons in the gene of interest were analyzed along with 20 base pairs of flanking intronic sequence. Genetic testing was most often performed on ABCA4, CTRP5, ELOV4, BEST1, CRB1, and PRPH2. Pathogenicity of novel sequence changes was predicted by PolyPhen2 and sorting intolerant from tolerant (SIFT). Results: Of the 225 genetic tests performed, 150 were for recessive IRD, and 75 were for dominant IRD. A positive molecular diagnosis was made in 70 (59%) of probands with recessive IRD and 19 (26%) probands with dominant IRD. Analysis confirmed 12 (34%) of individuals as carriers of familial mutations associated with IRD. Thirty-two novel variants were identified; among these, 17 sequence changes in four genes were predicted to be possibly or probably damaging including: ABCA4 (14), BEST1 (2), PRPH2 (1), and TIMP3 (1). Conclusions: Targeted analysis of clinically suspected genes in 225 subjects resulted in a positive molecular diagnosis in 26% of patients with dominant IRD and 59% of patients with recessive IRD. Novel damaging mutations were identified in four genes. Single gene screening is not an ideal method for diagnostic testing given the phenotypic and genetic heterogeneity among IRD cases. High-throughput sequencing of all genes associated with retinal degeneration may be more efficient for molecular diagnosis