Design and Synthesis of Thiazole Scaffold-Based Small Molecules as Anticancer Agents Targeting the Human Lactate Dehydrogenase A Enzyme.

A new series of thiazole central scaffold-based small molecules of hLDHA inhibitors were designed using an in silico approach. Molecular docking analysis of designed molecules with hLDHA (PDB ID: 1I10) demonstrates that Ala 29, Val 30, Arg 98, Gln 99, Gly 96, and Thr 94 possessed strong interaction with the compounds. Compounds 8a, 8b, and 8d showed good binding affinity (-8.1 to -8.8 kcal/mol), whereas an additional interaction of NO2 at the ortho position in compounds 8c with Gln 99 through hydrogen bonding enhanced the affinity to -9.8 kcal/mol. Selected high-scored compounds were synthesized and screened for hLDHA inhibitory activities and in vitro anticancer activity in six cancer cell lines. Biochemical enzyme inhibition assays showed the highest hLDHA inhibitory activity observed with compounds 8b, 8c, and 8l. Compounds 8b, 8c, 8j, 8l, and 8m depicted significant anticancer activities, exhibiting IC50 values in the range of 1.65-8.60 µM in HeLa and SiHa cervical cancer cell lines. Compounds 8j and 8m exhibited notable anticancer activity with IC50 values of 7.90 and 5.15 µM, respectively, in liver cancer cells (HepG2). Interestingly, compounds 8j and 8m did not induce noticeable toxicity in the human embryonic kidney cells (HEK293). Insilico absorption, distribution, metabolism, and excretion profiling demonstrates that the compounds possess drug-likeness, and results may pave the way for the development of novel thiazole-based biologically active small molecules for therapeutics.

Enhancement of maternal-neonatal bonding during the COVID-19 pandemic: A quality improvement initiative.

AIM: The COVID-19 pandemic adversely affected the essential care of newborns. In a tertiary care hospital in India, all COVID-19 suspect post-natal mothers awaiting COVID results were transferred to a ward shared with symptomatic COVID suspect female patients from other clinical specialities, due to shortage of space and functional health workforce. Babies born to COVID-19 suspect mothers were moved to a separate ward with a caretaker until their mothers tested negative. Due to shortage of beds and delay in receiving COVID results, mothers and babies were often discharged separately 2-3 days apart to their home. This deprived babies of their mother's milk and bonding. We, therefore, undertook a quality improvement (QI) initiative aiming to improve rooming-in of eligible COVID-19 suspect mother-newborn dyads from 0% to more than 90% over a period of 6 weeks. METHODS: A QI team was formed which ran multiple Plan-Do-Study-Act cycles. The results were reviewed at regular intervals and interventions were adopted, adapted or abandoned. These included advocacy, rearrangement of wards, counselling of mothers and caretakers regarding infection prevention practices and coordination between labour room, post-natal ward and nursery staff. RESULTS: An improvement in rooming-in from 0% to more than 90% was achieved. CONCLUSION: QI methodology is a systematic approach in addressing and solving unexpected unforeseen problems effectively.

Rational use of antibiotics for major elective gynaecological and obstetrical surgical procedures: quality improvement journey from a tertiary care public facility.

BACKGROUND: Antibiotic resistance is a global problem. Irrational use of antibiotics is rampant. Guidelines recommend administration of single dose of antibiotic for surgical antimicrobial prophylaxis (SSAP) for elective obstetrical and gynaecological surgeries. However, it is not usually adhered to in practice. Majority of women undergoing elective major gynaecological surgeries and caesarean sections in the department of obstetrics and gynaecology of our tertiary level heavy case load public health facility were receiving therapeutic antibiotics (for 7-10 days) instead of recommended SSAP. Our aim was to increase the SSAP in our setting from a baseline 2.1% to more than 60% within 6 months. METHODS: After root cause analysis, we formulated the departmental antimicrobial policy, spread awareness and sensitised doctors and nursing officers regarding antimicrobial resistance and asepsis through lectures, group discussions and workshops. We initiated SSAP policy for elective major surgeries and formed an antimicrobial stewardship team to ensure adherence to policy and follow processes and outcomes. The point of care quality improvement (QI) methodology was used. Percentage of patients receiving SSAP out of all low-risk women undergoing elective surgery was the process indicator and percentage of patients developing surgical site infection (SSI) of all patients receiving SSAP was the outcome indicator. The impact of various interventions on these indicators was followed over time with run charts. RESULTS: SSAP increased from a baseline 2.1%-67.7% within 6 months of initiation of this QI initiative and has since been sustained at 80%-90% for more than 2 years without any increase in SSI rate. CONCLUSION: QI methods can rapidly improve the acceptance and adherence to evidence-based guidelines in a busy public healthcare setting to prevent injudicious use of antibiotics.

Incidence, Management and Outcomes in Women Undergoing Peripartum Hysterectomy in a Tertiary Care Centre in India.

Background Peripartum hysterectomy (PRH) is the surgical removal of the uterus performed in obstetrical complications such as uncontrolled postpartum haemorrhage (PPH), unrepairable uterine rupture, and sepsis. Its incidence has increased in recent years. The objective of this study was to review all the cases of PRH in a tertiary care teaching hospital over three years (January 2017-December 2019) to determine its incidence and analyse clinico-demographic characteristics in these women. Method All women undergoing PRH from January 2017 to December 2019 were included in the study. Data were collected retrospectively from medical records, of patients who underwent a PRH at the time of delivery, or within 24 hours, or performed any time before discharge from the same hospitalization and obstetric event. The total number of deliveries including caesarean and vaginal deliveries were recorded. Main outcome measures were the incidence of PRH, indication for hysterectomy, management option used, maternal outcomes (PPH, bladder injury and maternal death) and fetal outcomes (stillbirth). Results There were a total of 3904,4 deliveries; 27,337 vaginal and 11,697 caesarean sections in three years. A total of 50 patients underwent a PRH. The incidence of PRH in our study was 1.3 per 1,000 deliveries and 3.5/1,000 caesareans, respectively. PRH was found to be more common following cesarean sections than vaginal deliveries (odds ratio 22.86 [95% CI: 8.16 to 63.98]). Morbid adherent placenta (MAP) (n=30, 62%) was the most common indications of PRH. Seven (15%) women had PRH due to uterine rupture. Twenty-seven women of the 30 women (90%) with the MAP had a previous caesarean delivery. The case fatality rate per hysterectomy was 4%. Stillbirth rate (SBR: n=8,16%) among women having PRH was seven-fold higher than overall SBR in our country. Conclusion There has been a rise in MAP as an indication of PRH in our study for a decade in comparison to uterine atony. Caesarean delivery is a significant risk factor for PRH. Previous caesarean section and major placenta previa were common occurring obstetric risk factors present in the MAP in our cohort. Our maternal mortality in PRH was low and the stillbirth rate was high when compared with national data.

Stillbirths and the COVID-19 pandemic: Looking beyond SARS-CoV-2 infection.

OBJECTIVE: To study the impact of the COVID-19 outbreak and subsequent lockdown on the incidence, associated causes, and modifiable factors of stillbirth. METHODS: An analytical case-control study was performed comparing stillbirths from March to September 2020 (cases) and March to September 2019 (controls) in a tertiary care center in India. Modifiable factors were observed as level-I, level-II, and level-III delays. RESULTS: A significant difference in the rate of stillbirths was found among cases (37.4/1000) and controls (29.9/1000) (P = 0.045). Abruption in normotensive women was significantly higher in cases compared to controls (P = 0.03). Modifiable factors or preventable causes were noted in 76.1% of cases and 59.6% of controls; the difference was highly significant (P < 0.001, relative risk [RR] 1.8). Level-II delays or delays in reaching the hospital for delivery due to lack of transport were observed in 12.7% of cases compared to none in controls (P < 0.006, RR 47.7). Level-III delays or delays in providing care at the facility were observed in 31.3% of cases and 11.5% of controls (P < 0.001, RR 2.7). CONCLUSION: Although there was no difference in causes of stillbirth between cases and controls, level-II and level-III delays were significantly impacted by the pandemic, leading to a higher rate of preventable stillbirths in pregnant women not infected with COVID-19.

Conservative management of cervical pregnancy: a report of two cases.

BACKGROUND: Cervical ectopic pregnancy is a rare form of ectopic pregnancy. Treatment of this clinical condition is controversial. Previously, hysterectomy was the mainstay of treatment, but in the recent past various conservative management approaches have been applied to preserve fertility. We report 2 such cases in which conservative approach was implemented. CASES: Case 1 was an early, nonviable cervical pregnancy in which systemic methotrexate was used with success. Case 2 was a viable, advanced cervical pregnancy (at 11 weeks) in which a combination of uterine artery embolization and systemic methotrexate was used. Ultimately this patient required hysterectomy due to sudden massive hemorrhage on day 22 of intervention. CONCLUSION: On reviewing our results and the literature, we conclude that uterine artery embolization with methotrexate is effective in reducing the ectopic cervical mass. However, there is always a risk of hemorrhage, which can be treated by either repeat uterine artery embolization alone or uterine artery embolization followed by curettage. Hysterectomy should be the last resort if all conservative methods fail.

Bionanowhiskers from jute: preparation and characterization.

Bionanowhiskers were extracted from jute by acid hydrolysis. At first cellulose microfibrils were formed by alkali treatment. Addition of an acid to the microfibrils triggered the formation of cellulose nanowhiskers. These were characterized by using different techniques viz. Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetry (TGA), scanning electron microscopy (SEM) and atomic force microscopy (AFM). In the FTIR study, absence of peak at 1738 cm(-1) indicated removal of hemicellulose. The rod like morphology of the nanowhiskers (length - 550±100 nm, width - 77±30 nm) was observed after 1h of acid hydrolysis, whereas further increase in time resulted in triangular shape morphology. Acid hydrolysis increased crystallinity, but decreased the temperature corresponding to major degradation (T(max)) and onset of degradation (T(i)). These bionanowhiskers might be useful as reinforcing element in nanocomposites.

Influence of phosphorus and pH on the fungicidal potential of Anabaena strains.

The genus Anabaena is known to be a rich source of bioactive metabolites, but the biocontrol potential of this genus, mediated through hydrolytic enzymes is less investigated. In our investigation, five Anabaena strains - A. laxa RPAN8, A. iyengarii RPAN9, A. variabilis RPAN59 and A. oscillarioides RPAN69 (with A. variabilis RPAN16 serving as negative control) were evaluated in time course studies involving incubation under three levels of phosphorus and pH conditions. Total chlorophyll, proteins, chitosanase, endoglucanase and CMCase activity were measured and inhibition assayed against phytopathogenic fungi. The four weeks old RPAN69 culture showed significantly higher chlorophyll which was 41% higher than control. This was also linked with an enhancement of 18.26% and 9.18% in chitosanase and CMCase activity respectively over control in the treatment involving half dose of phosphorus. Chlorophyll and CMCase activity showed a high degree of correlation with highest values at pH 9.5. A pH of 5.5 was the most suitable condition for the maximum activity of chitosanase for all the strains except RPAN16. The strains RPAN8 and RPAN9 showed the highest activity of endoglucanase at pH 5.5 while the other strains exhibited maximum activity at pH 7.5. This study provides insight into the role of P and pH in modulating fungicidal activity in different Anabaena strains, which can be valuable for enhancing their efficiency as a biocontrol agent.

Bioefficacy of novel cyanobacteria-amended formulations in suppressing damping off disease in tomato seedlings.

Biological control of plant pathogens is receiving increasing relevance, as compared to chemical methods, as they are eco-friendly, economical and indirectly improve plant quality and yield attributes. An investigation was undertaken to evaluate the potential of antagonistic cyanobacteria (Anabaena variabilis RPAN59 and A. oscillarioides RPAN69) fortified formulations for suppressing damping off disease in tomato seedlings challenged by the inoculation of a fungal consortium (Pythium debaryanum, Fusarium oxysporum lycopersici, Fusarium moniliforme and Rhizoctonia solani). Treatment with A. variabilis amended formulations recorded significantly higher plant growth parameters, than other treatments, including biological control (Trichoderma formulation) and chemical control (Thiram-Carbendazim). The A. variabilis amended compost-vermiculite and compost formulations exhibited 10-15 % lower disease severity and 40-50 % higher values than chemical and biological control treatments in terms of fresh weight and height of the plants. In future, in depth analyses regarding the mechanism involved in biocontrol by cyanobacteria and evaluation of these formulations under field conditions are proposed to be undertaken.

Modulation of fungicidal potential of Anabaena strains by light and temperature.

The regulation of fungicidal and hydrolytic enzyme activity was investigated in a set of cyanobacterial strains belonging to the genus Anabaena (Anabaena laxa RPAN8, Anabaena iyengarii RPAN9, Anabaena variabilis RPAN59 and Anabaena oscillarioides RPAN69), with A. variabilis RPAN16 serving as negative control. Time course studies undertaken with cultures incubated under different light and temperature conditions revealed enhancement in growth and fungicidal activity under continuous light (CL) and light dark (LD, 16:8) conditions and temperature of 30 °C and 40 °C. A significant increase of 3-18 % in chitosanase activity was recorded in all the 4-week-old cultures under CL condition and at 40 °C. Endoglucanase activity of RPAN8 and 9 was twofolds higher than the other strains under all light/dark conditions and temperature in the 4-week-old cultures, while continuous dark (CD) enhanced CMCase activity in RPAN69. This study provided useful information regarding the most suitable conditions of light and temperature for maximizing hydrolytic enzyme activity and fungicidal activity, as a prelude to their effective use as biocontrol agents.

Time resolved emission studies of Ag-adenine-templated CdS (Ag/CdS) nanohybrids.

Ag-adenine-templated CdS (Ag/CdS) nanohybrids have been synthesized and characterized by transmission electron microscopy, selected area electron diffraction, x-ray diffraction, and optical, fluorescence and time resolved emission spectroscopy. Adenine serves as an effective matrix for the stabilization of Ag/CdS through interaction of N(1), N(3) and -NH(2) with Ag. The amount of Ag in the nanohybrid is observed to influence the organization of the Ag and CdS phase in the composite and also modifies the nature of electronic transition in CdS. For the nanohybrid containing a molar ratio of 0.1 of Ag/ CdS, CdS nanoparticles (2.5 nm) surround the Ag (6.5 nm) core. The excitation of these particles by 340 nm light, where the absorption due to the Ag phase in the nanohybrid is negligibly small, results in the enhancement of fluorescence by a factor of 7 compared to that of bare CdS. For the particles containing a molar ratio of Ag/CdS of unity, bigger clusters (14 nm) are produced causing the quenching of emission of CdS. In time resolved emission spectroscopy the spectral shift from 415 nm (3.0 eV) to 550 nm (2.26 eV) monitored over a period of 1-220 ns is understood by the relaxation of charge within the surface states of varied energy from 180 to 370 eV. The observed changes in fluorescence behavior in terms of intensity, lifetime and spectral shift are understood in terms of electronic interaction between Ag and CdS phases. The manipulation of electronic and fluorescence properties in these nanohybrids could be exploited for optoelectronic, molecular-recognition and sensing applications.

Evaluation of fungicidal activity of extracellular filtrates of cyanobacteria--possible role of hydrolytic enzymes.

A set of seventy axenised and unicyanobacterial isolates belonging to the genus Anabaena were evaluated for biocidal activity against a set of phytopathogenic fungi. Among them, 35 Anabaena strains showed zone of inhibition against one or more fungi. The extracellular filtrates from 4 and 8 weeks old cultures of these Anabaena strains were further evaluated in terms of hydrolytic enzymes, proteins and IAA employing standard methods. Significant differences were also observed among the strains in terms of their FPase, chitosanase and xylanase activity, while low and relatively similar values of CMCase, cellobiase and protease activity were recorded in the strains analyzed. IAA production was also observed in all the strains. Comparative evaluation of activity of hydrolytic enzymes and antifungal activity revealed that such enzymes may contribute to the fungicidal activity of the cyanobacterial strains, besides other bioactive compounds, including IAA, which are established promising traits for biocontrol agents. This study is a first time report on the production of hydrolytic enzymes by these oxygenic photosynthetic prokaryotes, which can be potential candidates for the development of biocontrol agent(s) against selected phytopathogenic fungi.

Effects of arbuscular mycorrhiza and phosphorus application on artemisinin concentration in Artemisia annua L.

Annual wormwood (Artemisia annua L.) produces an array of complex terpenoids including artemisinin, a compound of current interest in the treatment of drug-resistant malaria. However, this promising antimalarial compound remains expensive and is hardly available on the global scale. Synthesis of artemisinin has not been proved to be feasible commercially. Therefore, increase in yield of naturally occurring artemisinin is an important area of investigation. The effects of inoculation by two arbuscular mycorrhizal (AM) fungi, Glomus macrocarpum and Glomus fasciculatum, either alone or supplemented with P-fertilizer, on artemisinin concentration in A. annua were studied. The concentration of artemisinin was determined by reverse-phase high-performance liquid chromatography with UV detection. The two fungi significantly increased concentration of artemisinin in the herb. Although there was significant increase in concentration of artemisinin in nonmycorrhizal P-fertilized plants as compared to control, the extent of the increase was less compared to mycorrhizal plants grown with or without P-fertilization. This suggests that the increase in artemisinin concentration may not be entirely attributed to enhanced P-nutrition and improved growth. A strong positive linear correlation was observed between glandular trichome density on leaves and artemisinin concentration. Mycorrhizal plants possessed higher foliar glandular trichome (site for artemisinin biosynthesis and sequestration) density compared to nonmycorrhizal plants. Glandular trichome density was not influenced by P-fertilizer application. The study suggests a potential role of AM fungi in improving the concentration of artemisinin in A. annua.