Analytical quality by design approach for the development of high-performance liquid chromatography method for simultaneous analysis of metformin and sitagliptin in the presence of major degradation products.

An analytical quality by design-based high-performance liquid chromatography method for determining metformin (MET) and sitagliptin (SIT) in stress-degraded samples was developed and validated. The analytical target profile and risk assessment-driven critical method variables, for example, pH, % aqueous, and buffer concentration, were studied for their effect on method responses of retention time and resolution using a central composite design. The correlation regression coefficient was more than 0.8, and variables interaction was significant on method responses with curvature effect. The method operable design region afforded an aqueous range of 55%-70% and an ortho-phosphoric acid buffer of 0.1% with a pH of 3.0-4.0 as a robust region for the suitable method performance characteristics. The separation of MET and SIT from their degradants (m/z 85.0509; m/z 193.0694) on the C8 column was achieved using a mobile phase consisting of 0.1% ortho-phosphoric acid and methanol (60:40% v/v; pH 3.0). The optimized method eluted MET and SIT at 4.3 ± 0.2 and 7.1 ± 0.2 min, respectively, with acceptable specificity and resolution. The linearity ranges of 25-250 µg/mL (r2 : 0.9982) and 5-50 µg/mL (r2 : 0.9989) was established for MET and SIT, respectively. The % recovery (98.81%-102.17%), precision (0.55%-1.65%), and robustness study for method variables were acceptable.

Mechanistic insight into the role of mevalonate kinase by a natural fatty acid-mediated killing of Leishmania donovani.

We evaluated the anti-leishmanial efficacy of different saturated medium-chain fatty acids (FAs, C8-C18) where FA containing C8 chain, caprylic acid (CA), was found to be most potent against Leishmania donovani, the causative agent for visceral leishmaniasis (VL). Different analogs of CA with C8 linear chain, but not higher, along with a carboxyl/ester group showed a similar anti-leishmanial effect. Ergosterol depletion was the major cause of CA-mediated cell death. Molecular docking and molecular dynamic simulation studies indicated the enzyme mevalonate kinase (MevK) of the ergosterol biosynthesis pathway as a possible target of CA. Enzyme assays with purified recombinant MevK and CA/CA analogs confirmed the target with a competitive inhibition pattern. Using biochemical and biophysical studies; strong binding interaction between MevK and CA/CA analogs was established. Further, using parasites with overexpressed MevK and proteomics studies of CA-treated parasites the direct role of MevK as the target was validated. We established the mechanism of the antileishmanial effect of CA, a natural product, against VL where toxicity and drug resistance with current chemotherapeutics demand an alternative. This is the first report on the identification of an enzymatic target with kinetic parameters and mechanistic insights against any organism for a natural medium-chain FA.

An exclusive hand protection device made of fused deposition modelling process using poly (lactic acid) polymer.

The world has been hit hardest by a type of Severe Acute Respiratory Syndrome (SARS) virus called corona. This vulnerability did not leave all the superpowers in the world. The virus, is spread through coughs and sneezes. It is also spread by touching objects or parts of an infected person. For this reason, it is considered an infectious disease. It is very difficult to protect ourselves from such a contagious disease. But the World Health Organization (WHO) says there are certain guidelines to follow. The first way is to prevent the spread of COVID-19 social infection by adhering to social spaces (i.e. 2 m intervals). Second, we can protect ourselves by using appropriate safety equipment. We can also protect ourselves from the COVID-19 virus by using sanitizers or soaps. So this study focuses on a 3D-printed hand protection device for COVID- 19 infection prevention. The design, fabrication, and testing of the gadget were all successful. Thus, a low-cost and efficient device made by eco-friendly Poly Lactic Acid (PLA) polymer material and may be utilized to operate some potentially susceptible and highly infectious surfaces in a variety of public areas, where touching and using particular components is common and frequent (eg door handles, electrical switches, ATM machines).

A critical review of an additive manufacturing role in Covid-19 epidemic.

In 2019, a massive and deadly coronavirus pandemic known as the COVID-19 pandemic has swept through more than 180 nations, causing a massive strain on already overtaxed health systems around the globe. Global demand for medical equipment has put a strain on traditional manufacturing methods, resulting in the need for an efficient, low-cost, and speedy mode of production. Additive manufacturing, or 3D printing, has been used by manufacturers to bridge the gap and enhance the production of medical products. Some designs that had been previously or conventionally fabricated have been revised to meet the 3D printing requirement for combating COVID-19. A variety of designs were created, and they are now in use in hospitals by patients and healthcare professionals. However, because some gadgets must adhere to rigorous standards, it is possible that some items will not meet these requirements. As a result, in order to protect the health of the user, it is necessary to understand each gadget, its usage, and industry standards. An investigation of the usage of additive manufacturing during the COVID-19 epidemic is presented in this paper. It brings together the manufacturers of a variety of 3D-printed products, including face shields, face masks, valves, nasopharyngeal swabs, and others, to debate their application and regulatory concerns in the medical field. The primary shortcoming of technology, discussed in reference to the next pandemic, is addressed here. It also looks at some of the ways that additive manufacturing could be used in the future during an emergency.

Arsenic: a Culpable Element and a Possible Menace for HIV/AIDS Patients.

Arsenic contamination has long been recognized as one of the most harmful environmental pollutants resulting from anthropogenic activity. Apart from being an environmental toxicant or pollutant, this culpable heavy metal also has detrimental effects on human health. People throughout the world are exposed to arsenic (As) mostly through polluted drinking water. Acute inorganic arsenic (iAs) poisoning causes nausea, vomiting, stomach discomfort, and severe diarrhea. As on long-term exposure is a potent carcinogen, characterized by IARC (International Agency for Research on Cancer). As levels are high mainly in Gangetic regions due to which the people living around are suffering the consequences. The carcinogenicity of As is well established but the immunotoxicity caused by it is still unknown. Some animal model supports the toxicity of As in the immune system as well, but in humans, mainly suffering from human immunodeficiency virus (HIV), it is not well established. iAs suppresses the immune system by acting on different targets and exacerbating infections. Although animal studies have demonstrated that arsenic trioxide (As2O3) reduces viral rebound and restores CD4 + count in vivo when coupled with antiretroviral medications, elemental AS may have devastating effects on the immune system of HIV patients, making them more prone to opportunistic infections (OIs). It is well known that in later stages of HIV infection, neurological problems also complicate the conditions such as cognitive impairment and AIDS dementia complex (ADC). Along with immunotoxicity, As has the potential to damage HIV patients' brains. This article addresses the immunotoxicity of arsenic and exacerbations caused by it, along with the neurotoxicity, particularly in HIV patients residing near the Gangetic belt.

Detection of Orientia tsutsugamushi in Novel Trombiculid Mite Species in Northern Tamil Nadu, India: Use of Targeting the Multicopy traD Gene.

Detection of Orientia tsutsugamushi DNA in a trombiculid mite chigger species suggests that it might be a potential vector of scrub typhus in an endemic area. Over a period of 20 mo, 85 rats were trapped, 57 had chiggers that were identified by standard morphometric techniques. The chigger pools were assessed by performing PCR assays targeting fragments of the single-copy genes 56 kDa type-specific antigen gene (TSA56) by nested PCR and the 47 kDa (htrA) quantitative real-time PCR (qPCR). The novel traD SYBR green assay that detects a multicopy gene was also performed. In total, 27 chigger pools were positive by traD qPCR, of which only 7 were positive by 47 kDa qPCR and in 3 of these, 56 kDa gene was amplified by nested PCR. Orientia tsutsugamushi-specific DNA was detected in Ascoschoengastia spp., Schoengastiella ligula, Leptotrombidium rajasthanense, Leptotrombidium deliense, and Leptotrombidium jayawickremei chigger pools. Therefore, they could be potential vectors of scrub typhus in Southern India. The three 56 kDa sequences belonged to TA716 genotype and Kato genogroup. Further studies are needed to confirm these chigger species as scrub typhus vectors in Northern Tamil Nadu.

Can MRI help assess aggressiveness of endometrial cancer?

AIM: To identify potential magnetic resonance imaging (MRI) biomarkers to predict the aggressiveness of endometrial cancer. MATERIALS AND METHODS: Seventy-one patients with endometrial cancer who underwent MRI staging were analysed retrospectively. The signal intensity (SI) of the tumours was assessed on sagittal T2-weighted imaging (WI) and sagittal T1WI sequences). The depth of myometrial invasion, tumour grade and subtype, lymphovascular invasion, and microsatellite stability status were assessed histopathologically, and these findings were compared with MRI findings using logistic regression. The log-rank test was used to assess differences in survival among groups defined by different MRI measurements. RESULTS: Tumours with qualitative higher signal than that of normal myometrium on the late T1WI DCE image sequences were more likely to have lymphovascular space invasion (p<0.001). Tumours that had a higher SI tumour ratio (T1 post-contrast arterial/T1 precontrast) had a higher chance of being microsatellite stable (odds ratio 2.36). The SI ratio of the tumour to the myometrium showed that lower T2 tumour/T2 myometrial ratio correlated with ≥50% depth of myometrial invasion as determined by imaging (p=0.006). Endometrial tumours showing a SI of >209 on delayed T1WI sequences had longer recurrence-free survival than those with tumours showing a SI ≤209 (p=0.014). Tumour subtype and grade were not associated with MRI findings. CONCLUSION: The SI of endometrial cancer on MRI may be used to predict the aggressiveness of the tumour and microsatellite stability status. Further studies are needed to confirm these findings.

Combination therapy with topotecan, paclitaxel, and bevacizumab improves progression-free survival in recurrent small cell neuroendocrine carcinoma of the cervix.

OBJECTIVES: To assess if the combination of topotecan, paclitaxel, and bevacizumab (TPB) was active in recurrent SCCC and to compare the survival of patients with SCCC who received TPB to a group of women with SCCC who did not receive this regimen. METHODS: We retrospectively analyzed women with recurrent SCCC who received chemotherapy as primary therapy. Women treated with TPB for first recurrence were compared to women treated with non-TPB chemotherapy. RESULTS: Thirteen patients received TPB, and 21 received non-TPB chemotherapy, most commonly platinum with or without a taxane. Median progression-free survival (PFS) was 7.8months for TPB and 4.0months for non-TPB regimens (hazard ratio [HR] 0.21, 95% CI 0.09-0.54, P=0.001). Median overall survival (OS) was 9.7months for TPB and 9.4months for non-TPB regimens (HR 0.53, 95% CI 0.23-1.22, P=0.13). Eight women (62%) who received TPB versus four (19%) who received non-TPB regimens were on treatment for >6months (P=0.02), and four patients (31%) in the TPB group versus two (10%) in the non-TPB group were on treatment for >12months (P=0.17). In the TPB group, three patients (23%) had complete response, two (15%) had complete response outside the brain with progression in the brain, 3 (23%) had a partial response, 2 (15%) had stable disease, and 3 (23%) had progressive disease. CONCLUSIONS: These findings indicate that TPB for recurrent SCCC significantly improved PFS over non-TPB regimens, and trends towards improved OS. Furthermore, a significant number of patients had a durable clinical benefit.

Lipemia Retinalis due to Secondary Hyperlipidemia in Type 1 Diabetes Mellitus.

Lipemia retinalis is a rare and asymptomatic condition which occurs when high levels of triglycerides and chylomicrons are present in blood. We report a rare case of secondary hyperlipoproteinemia in a 27 year old type 1 diabetes patient who presented with diabetic ketoacidosis and this peculiar ocular manifestation. The fundoscopic abnormality and creamy white serum cleared as the level of chylomicrons in the plasma dropped with intensive insulin therapy.

Wilson's Disease with Systemic Lupus Erythematosus.

Case reports of Wilson's disease occurring in combination with SLE are rarely reported in literature. Drug induced lupus have been observed in patients taking D-penicillamine for Wilson's disease. Here we report a case from Coimbatore Medical College hospital, who presented with fever and neuropsychiatric symptoms as the initial manifestation and found to have both SLE and Wilson's disease on subsequent evaluation.

Cerebral Sinus Venous Thrombosis as a Rare Complication of Primary Varicella Zoster Virus Infection.

We present the case of a 23 year old with acute onset left hemiparesis and meningeal irritation, associated with recent history of chickenpox 15 days prior. Varicella-IgG and IgM was positive in the CSF and blood along with reduced serum/CSF ratios of VZV immunoglobulins. MRV showed thrombosis (CVT) of superior sagittal, transverse, right sigmoid sinuses with haemorrhagic infarct in right frontoparietal region. Patient responded well to intravenous heparin, Acyclovir and oral anticoagulant therapy.

Idiopathic CD4 Lymphocytopenia.

We report a 42 year old male who was an interesting case of "idiopathic CD4 lymphocytopenia"(ICL) in a non-HIV adult with-extra pulmonary tuberculosis along with the diffuse splenic calcification and whose symptoms improved with Antitubercular treatment. He was found to have low CD4 counts on two occasions.

Pulmonary Thromboembolism Presenting as Multiple Pulmonary Cavities.

A 38 years old male patient presented to the emergency department with acute severe retrosternal chest pain and was found to have pulmonary thromboembolism. The patient developed new necrotic lung cavities each day due to the dissemination of emboli from the thrombus. The cause of thrombus was found to be protein C deficiency, which is an inherited thrombophilia. The interesting features in this case are multiple lung cavitations and high grade fever, which was attributed to pulmonary thromboembolism. The patient improved symptomatically with anticoagulants and antiplatelets.

Transient ST Segment Elevation following Indian Red Scorpion Sting with Non-Occlusive Right Coronary Artery Disease.

A 50 yr old female developed transient ST segment elevation due to an acute coronary vasospasm following a Indian Red Scorpion sting and the angiogram revealed 50% occlusion in right coronary artery (RCA).The possible mechanism is that the sympathetic over-activity could have aggravated the occlusion in the RCA as the RCA is more commonly prone to spasm compared to the other coronary arteries.

Is MRI helpful in assessing the distance of the tumour from the internal os in patients with cervical cancer below FIGO Stage IB2?

AIM: To determine the ability of magnetic resonance imaging (MRI) in detecting tumour-free margins from the internal os (IO). MATERIALS AND METHODS: A database search yielded 79 women with early-stage cervical cancer who underwent radical hysterectomy and preoperative MRI. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and accuracy of MRI in assessment of ≤5 and >5 mm IO involvement were calculated with histopathological surgical specimen findings considered to be the reference standard. A main and subset analysis was performed. The subset analysis included only those patients who would have been considered for radical trachelectomy. RESULTS: For predicting a distance between the tumour and the IO of ≤5 mm, MRI had a sensitivity of 73%, a specificity of 98.3%, a PPV of 95%, a NPV of 88.1%, and an accuracy of 89.8% for the main analysis, and sensitivity of 81.8%, a specificity of 93.2% a PPV of 69.2% a NPV of 96.5% and an accuracy of 91.4% for the subset analysis. CONCLUSION: MRI has high specificity, NPV, and accuracy in detecting tumour from the IO, making MRI suitable for treatment planning in patients desiring trachelectomy to preserve fertility.

Stability-indicating RP-HPLC Method for the Simultaneous Determination of Sitagliptin and Simvastatin in Tablets.

A new stability-indicating high-performance liquid chromatographic method for simultaneous analysis of sitagliptin and simvastatin in pharmaceutical dosage form was developed and validated. The mobile phase consisted of methanol and water (70:30, v/v) with 0.2 % of n-heptane sulfonic acid adjusted to pH 3.0 with ortho phosphoric acid was used. Retentions of sitagliptin and simvastatin were 4.3 min and 30.4 min, respectively with a flow rate of 1 ml/min on C8 (Qualisil BDS, 250×4.6 mm, 5 µ). Eluents were detected at 253 nm using photodiode diode array detector. The linear regression analysis data for the linearity plot showed correlation coefficient values of 0.9998 and 0.9993 for sitagliptin and simvastatin, with respective concentration ranges of 20-150 µg/ml and 8-60 µg/ml. The relative standard deviation for inter-day precision was lower than 2.0%. The assay of sitagliptin and simvastatin was determined in tablet dosage form was found to be within limits. Both drugs were subjected to a variety of stress conditions such as acidic, basic, oxidation, photolytic, neutral and thermal stress in order to achieve adequate degradation. Results revealed that considerable degradation was found in all stress conditions except oxidative degradations. The method has proven specificity for stability indicating assay method.

Evaluation of metformin hydrochloride in Wistar rats by FTIR-ATR spectroscopy: A convenient tool in the clinical study of diabetes.

INTRODUCTION: The IR absorption patterns (in cm(-1)) provide the basis to distinguish among the constituents and to separately quantify as well as qualify them and they possess many advantages such as very small sample volume requirement, good precision over entire physiological range, avoid of costly disposables, wealth of information from a single spectral measurement. The efficacy of anti-diabetic drug metformin hydrochloride as used to treat diabetic-induced Wistar rats and their sera were analyzed by FT-IR (ATR) in absorption mode. MATERIALS AND METHODS: The present work was attempted in the study of normal and antidiabetic regimen-treated rat blood samples using FTIR spectroscopy by the attenuated total reflectance (ATR) sampling technique. The biomolecule characteristics were measured as intensity ratio parameter (IRP) values and interpreted. RESULTS: To quantify the results three IRPs such as R1, R2 and R3 were calculated, respectively, for lipid, protein, and glucose. The glucose IRP value R3 showed, 0.3802, 0.3304, and 0.2847, respectively, for diseased, metformin-treated, and normal rats. CONCLUSION: The IRP values for glucose are compared to the glucose level obtained by using a glucometer. This study can be conveniently used in diagnostic procedures, patient compliance assessment, and efficacy evaluation of metformin hydrochlorides.

Emerging association between androgen deprivation therapy and male meningioma: significant expression of luteinizing hormone-releasing hormone receptor in male meningioma.

BACKGROUND: There is emerging data suggesting a potential risk for meningioma growth stimulation in patients on luteinizing hormone-releasing hormone (LHRH) analogs for prostate cancer. We examined the expression of LHRH receptor (LHRH-R), progesterone receptor (PR) and Ki67 labeling index (LI) in specimens from male meningioma (MM) and female meningioma (FM) patients. METHODS: A total of 24 MM and 24 FM paraffin blocks were retrieved from our institution between 1991 and 2008. Sections from the paraffin blocks were stained with mouse monoclonal antibodies against LHRH-R, PR and Ki67. All male patients had no previous history of prostate cancer (PCa) or previous history of hormone therapy. RESULTS: LHRH-R positivity was extensive in 92% of MM and 88% of FM samples, with both showing strong intensity (67% and 79%, respectively). PR was positive in 20 of 24 (83%) MM and 23 of 24 (96%) FM samples. MM is less likely to exhibit Ki67 LI >4% compared with FM. CONCLUSIONS: The majority of MM and FM samples were strongly positive for LHRH-R expression and PR expression. The emerging association of androgen deprivation therapy and meningioma growth should be recognized in urological practice. Caution should be taken when considering LHRH agonist administration for patients with PCa and concurrent meningioma or previous history of meningioma.

MRI is highly specific in determining primary cervical versus endometrial cancer when biopsy results are inconclusive.

AIM: To evaluate utility of magnetic resonance imaging (MRI) in determining the primary site of endometrial versus cervical tumours when the biopsy results are inconclusive. MATERIAL AND METHODS: Forty-eight patients who underwent a total hysterectomy for unknown primary adenocarcinoma of the uterus after endometrial and/or endocervical biopsies were included in the study. The 48 available pelvic MRI images were reviewed by two body radiologists independently and jointly to resolve discordance, blinded to any clinical and pathological information. The clinical information and histopathology were reviewed by a radiology fellow and a pathologist specializing is gynaecological oncology. The final surgical pathology was used as the reference standard to confirm the origin of the primary tumour. RESULTS: The radiologists correctly identified the primary sites in 85% of the cases (41/48). There was substantial agreement between the two readers (kappa statistics = 0.79). Both radiologists found that T2 and dynamic T1-weighted images (WI) were most helpful in making the diagnosis, and dynamic T1WI helped resolve problematic cases. The sensitivity and specificity for detecting endometrial and cervical cancer on MRI were 88% and 88% and 75% and 93%, respectively. CONCLUSION: MRI has high sensitivity and specificity in determining the origin of the primary endometrial versus cervical tumours when endometrial/endocervical curettage is inconclusive.