Anticancer potential of myricetin bulk and nano forms in vitro in lymphocytes from myeloma patients.

Evading apoptosis and chemo-resistance are considered as very important factors which help tumour progression and metastasis. Hence, to overcome chemo-resistance, there is an urgent requirement for emergence of more effective treatment options. Myricetin, a naturally occurring flavonoid, is present in various plant-derived foods and shows antitumour potential in different cancers. In the present in vitro study, results from the comet assay demonstrated that myricetin bulk (10 µM) and nano (20 µM) forms exhibited a non-significant level of genotoxicity in lymphocytes from multiple myeloma patients when compared to those from healthy individuals. Western blot results showed a decrease in Bcl-2/Bax ratio and an increase in P53 protein levels in lymphocytes from myeloma patients, but not in lymphocytes from healthy individuals. A significant increase in intracellular reactive oxygen species level was also observed, suggesting that regulation of apoptotic proteins triggered by myricetin exposure in lymphocytes from myeloma patients occurred through P53 and oxidative stress-dependent pathways. The potency of myricetin against lymphocytes from myeloma patients marks it a potential candidate to be considered as an alternative to overcome chemo-resistance in cancer therapies.

Ex vivo/in vitro protective effect of myricetin bulk and nano-forms on PhIP-induced DNA damage in lymphocytes from healthy individuals and pre-cancerous MGUS patients.

2-Amino-1-methyl-6-phenylimidazo [4,5-b]pyridine (PhIP) is a central dietary mutagen, produced when proteinaceous food is heated at very high temperatures potentially causing DNA strand breaks. This study investigates the protective potential of a well-researched flavonoid, myricetin in its bulk and nano-forms against oxidative stress induced ex vivo/in vitro by PhIP in lymphocytes from pre-cancerous monoclonal gammopathy of undetermined significance (MGUS) patients and those from healthy individuals. The results from the Comet assay revealed that in the presence of myricetin bulk (10 µM) and myricetin nano (20 µM), the DNA damage caused by a high dose of PhIP (100 µM) was significantly (P < 0.001) reduced in both groups. However, nano has shown better protection in lymphocytes from pre-cancerous patients. Consistent results were obtained from the micronucleus assay where micronuclei frequency in binucleated cells significantly decreased upon supplementing PhIP with myricetin bulk (P < 0.01) and myricetin nano (P < 0.001), compared to the PhIP treatment alone. To briefly determine the cellular pathways involved in the protective role of myricetin against PhIP, we studied gene expression of P53 and ATR kinase (ATM- and Rad3-related), using the real-time PCR technique.

ROS-induced oxidative damage in lymphocytes ex vivo/in vitro from healthy individuals and MGUS patients: protection by myricetin bulk and nanoforms.

We investigated the protective role of myricetin bulk and nanoforms, against reactive oxygen species (ROS)-induced oxidative stress caused by hydrogen peroxide and tertiary-butyl hydro peroxide in lymphocytes in vitro from healthy individuals and those from pre-cancerous patients suffering with monoclonal gammopathy of undetermined significance (MGUS). The change in intracellular reactive oxygen species was measured once cells were treated with myricetin bulk forms and nanoforms with and without either hydrogen peroxide or tertiary-butyl hydro peroxide co-supplementation. The direct and indirect antioxidant activity of myricetin was spectrofluometrically measured using the fluorescent dye 2',7'-dichlorofluorescin diacetate and using the Comet assay, respectively. Hydrogen peroxide (50 µM) and tertiary-butyl hydro peroxide (300 µM) induced a higher level of reactive oxygen species-related DNA damage and strand breaks. Addition of myricetin nanoform (20 µM) and bulk (10 µM) form could, however, significantly prevent hydrogen peroxide- and tertiary-butyl hydro peroxide-induced oxidative imbalances and the nanoform was more effective. Glutathione levels were also quantified using a non-fluorescent dye. Results suggest that myricetin treatment had no significant effect on the cellular antioxidant enzyme, glutathione. The current study also investigates the effect of myricetin on the induction of double-strand breaks by staining the gamma-H2AX foci immunocytochemically. It was observed that myricetin does not induce double-strand breaks at basal levels rather demonstrated a protective effect.

Analysis of barriers affecting Industry 4.0 implementation: An interpretive analysis using total interpretive structural modeling (TISM) and Fuzzy MICMAC.

The purpose of this study is to build a structural relationship model based on total interpretive structural modeling (TISM) and fuzzy input-based cross-impact matrix multiplication applied to classification (MICMAC) for analysis and prioritization of the barriers influencing the implementation of Industry 4.0 technologies. 10 crucial barriers that affect the deployment of Industry 4.0 techniques are identified in the literature. Also, the Fuzzy MICMAC approach is applied to classify the barriers. The importance of TISM over traditional interpretive structural modeling (ISM) is shown in this work. Results proved that the barriers, namely IT infrastructure, lack of cyber physical systems, and improper communication models, are identified as the most dependent barriers, and the barriers of lack of top management commitment and inadequate training are identified as the most driving barriers. This study makes it easier for decision-makers to take the necessary steps to mitigate the barriers. The bottom level of the TISM hierarchy is occupied by barriers that need more attention from top management in order to be effectively monitored and managed. This study explains the steps to execute TISM in detail, making it easy for researchers and practitioners to comprehend its principles.

Think fungus NOT just a crypto-meningitis in AIDS!

UNLABELLED: Extrapulmonary cryptococcosis has been defined as AIDS defining illness in HIV infected people. Cryptococcal meningitis is the commonest meningitis with advanced immune deficiency. Therefore clinicians ask for tests only for detection of cryptococci which may be misleading. A prospective study of suspected fungal meningitis with CSF fungal culture is carried out. MATERIAL AND METHODS: 70 ART naive cases of suspected fungal meningitis in HIV cases were subjected to CSF cytochemistry, smear exam and CSF fungal culture. RESULTS: The CSF culture was positive in 75.6% cases of these 21 were C. Neoformans as against 28 of Rhodotorula. In addition candida, aspergillus, geotrichum, trichosporon were isolated. CONCLUSION: Apart from c. neoformans, other fungi also cause meningitis. Each case of suspected fungal meningitis, may be subjected for CSF fungal culture for proper and adequate management. If facility for fungal culture is not available and if CSF smear shows evidence of fungal infection then standard therapy with Amphotericin may be instituted earlier to reduce mortality. This is the largest series isolating Rhodotorula from CSF in AIDS patients.

Entrepreneurial Interventions for crisis management: Lessons from the Covid-19 Pandemic's impact on entrepreneurial ventures.

This article investigates both the negative and positive impacts of a crisis on Entrepreneurial Ventures. The behaviour of Entrepreneurial Ventures during the COVID-19 pandemic crisis is studied by undertaking a Systematic Literature Review (SLR) using Bibliometrics of 154 related publications. After analyzing the literature, the behaviour of Entrepreneurial Ventures during the crisis is synthesized and presented as a Phenomenon Structure Diagram, which highlights a combination of Entrepreneurial Actions (EAs) and Entrepreneurial Orientations (EOs), with Entrepreneurial Supports (ESs) employed by them to manage the crisis. This combination of EAs and EOs with ESs is contingent on the surrounding environment, and they aid in minimizing the risk of failure as well as leverage new opportunities. We propose and develop a conceptual model that a combination of EAs and EOs with ESs, referred to as Entrepreneurial Interventions, can better manage a crisis or disaster and improve organizational resilience. This multi-disciplinary study contributes towards theory development that Entrepreneurial Interventions can be made as a crisis and disaster management strategy.

In-Vitro In-Vivo Correlation (IVIVC) of Inhaled Products Using Twin Stage Impinger.

In vitro dissolution testing as a form of quality control has become a necessity in the pharmaceutical industry. As such, the need to establish a method that investigates the in vitro dissolution profile of inhaled products should be taken into account. The prime focus in this study was to examine the in-vitro in-vivo correlation utilising a modified version of the Twin Stage Impinger and to promote an in vitro dissolution model by enhancing the Fine Particle Dose (FPD) collection method for dry powder inhalers. The Twin Impinger was modified by inserting a stainless steel membrane holder disk in the base of the lower chamber. The design, with optimum drug deposition, was adopted for the dissolution study of budesonide and salbutamol. Afterwards, the membrane holder system was placed in the bottom of the dissolution vessel. Phosphate buffer saline (PBS), simulated lung fluid (SLF, Gamble solution) and Phosphate buffer (PB) were used in the study. The paddle dissolution apparatus, containing 300 mL of the medium, was operated at 75 rpm paddle speed. Samples were collected at defined time intervals and analysed using a validated HPLC method. The largest proportion of the budesonide dose was dissolved in PBS compared to PB and SLF. This was due to the presence of surfactant (0.2% w/v polysorbate), which enhances the wettability and the solubility of the poorly soluble drug (budesonide). The similarity factors for PBS and PB were 47.6 and 69.7, respectively, using SLF as a reference, whereas the similarity factor for salbutamol dissolution between PB and SLF was 81.3, suggesting PB is a suitable substitute. Comparison using both the predicted and actual in vivo pharmacokinetics (PK) values of the two drugs, as well as the pattern of their Concentration-Time (c-t) profiles, showed good similarity, which gave an indication of the validity of this in vitro dissolution method.

Predecessors of the giant 1960 Chile earthquake.

It is commonly thought that the longer the time since last earthquake, the larger the next earthquake's slip will be. But this logical predictor of earthquake size, unsuccessful for large earthquakes on a strike-slip fault, fails also with the giant 1960 Chile earthquake of magnitude 9.5 (ref. 3). Although the time since the preceding earthquake spanned 123 years (refs 4, 5), the estimated slip in 1960, which occurred on a fault between the Nazca and South American tectonic plates, equalled 250-350 years' worth of the plate motion. Thus the average interval between such giant earthquakes on this fault should span several centuries. Here we present evidence that such long intervals were indeed typical of the last two millennia. We use buried soils and sand layers as records of tectonic subsidence and tsunami inundation at an estuary midway along the 1960 rupture. In these records, the 1960 earthquake ended a recurrence interval that had begun almost four centuries before, with an earthquake documented by Spanish conquistadors in 1575. Two later earthquakes, in 1737 and 1837, produced little if any subsidence or tsunami at the estuary and they therefore probably left the fault partly loaded with accumulated plate motion that the 1960 earthquake then expended.

Investigation of FSSW parameters on shear fracture load of AA6061 and copper alloy joints.

In this research, Friction stir spot welding (FSSW) is extensively employed to join dissimilar metals consisting of AA6061-T6 Aluminium Alloy and Commercial Copper Alloy. These alloys were friction stir spot welded using process parameters with the major impact, such as Dwell Time (DT), Rotational Speed (RS), Plunge Rate (PR) and Tool Diameter Ratio (D/d). Trail experiments have been carried out using Design of Experiments. Sound welded joints exhibiting a maximum shear fracture load of 4.79 kN were obtained at a RS of 2000 rpm, PR of 7 mm/min, DT of 25 s and D/d of 3.5, Balanced material flow around the pin was also observed at this condition, and it is evident that the superior metallurgical properties and geometric features at this condition results in the higher strength. Moreover, the joint also exhibited higher load-bearing capabilities due to an optimal hook height of 0.89 mm, hook width of 1.3 mm and hook initiation distance of 1.4 mm. The continuous formation of intermetallic compound such as ɤ2-Cu9Al4 in the Cu zone and ɳ2-CuAl in the Al zone may also enhance the shear fracture load.

An in vitro investigation into the protective and genotoxic effects of myricetin bulk and nano forms in lymphocytes of MGUS patients and healthy individuals.

The present study investigated the genoprotective and genotoxic effects of myricetin bulk (10 µM) and nano forms (20 µM) in the lymphocytes from pre-cancerous, monoclonal gammopathy of unknown significance (MGUS) patients and healthy individuals using the Comet and micronucleus assays. The study also evaluated the effect of myricetin on P53 expression levels, using the Western blot technique. Results showed that throughout the in-vitro treatment, lymphocytes from the patients group had higher levels of baseline DNA damage compared to the healthy group. Myricetin in both forms induced significant DNA damage, only at higher concentrations (>40 µM). The micronucleus assay showed a significant reduction (P < 0.01) in the frequency of micronuclei in mono-nucleated cells in the patient group treated with the nano form of myricetin at the non-toxic dose of 20 µM. There was a significant increase in both gene and protein P53 levels in lymphocytes isolated from healthy individuals and pre-cancerous patients. These results suggested a protective effect of myricetin and indicated its nutritional supplement potential for protection against cancer development among patients suffering from MGUS.

Data set on prediction of friction stir welding parameters to achieve maximum strength of AA2014-T6 aluminium alloy joints.

Statistical tools such as design of experiments (DoE), analysis of variance (ANOVA) were used to develop the empirical relationship, to predict the ultimate tensile strength of the joint at the 95% percent confidence level. Response surface graph and contour plots were constructed using response surface methodology (RSM) concept. From this investigation, it is found that the joint fabricated with a tool rotational speed of 1500 rpm, welding speed of 40 mm/min, tool tilt angle of 1.5° and tool shoulder diameter of 6 mm, exhibited maximum tensile strength of 380 MPa.

Restriction Fragment Length Polymorphism-based Genotyping of Toxoplasma gondii from Autopsy-Proven Cases of Acquired Immunodeficiency Syndrome-associated Cerebral Toxoplasmosis.

CONTEXT: Published data on genetic characterization of Toxoplasma gondii (T.gondii) from clinical cases of toxoplasmosis from India is lacking. AIMS: The present study was aimed at identifying genetic types of T. gondii in fatal cases of cerebral toxoplasmosis (CT) associated with HIV, from India. SETTINGS AND DESIGN: Archived tissues of CT were obtained postmortem from 25 acquired immunodeficiency syndrome patients between 2000 and 2014. SUBJECTS AND METHODS: Direct amplification of eight different loci, namely, SAG1, 5'-3'SAG2, Alt. SAG2, SAG3, BTUB, GRA6, C22-8, and L358 followed by restriction fragment length polymorphism was used to genotype the parasite. RESULTS: The canonical Types I, II, or III were not found in our study. More than 96% of the cases harbored atypical genotypes-likely recombinants of the canonical types; one case closely corresponded to Type II genotype. CONCLUSIONS: Thus, a majority of T. gondii causing CT in South India belonged to a noncanonical lineage. These nonarchetypal genotypes differed from the conventional Types I, II, and III and caused devastating severity in patients with CT in the background of HIV. These results are a step further to deciphering the population genetics of this important zoonotic parasitic infection in Indian patients, information that has thus far been lacking.

Nyctotherus sp. infection in pet turtle: a case report.

The present clinical case represents the successful therapeutic management of Nyctotherus sp. protozoan infection in turtles. Two pet turtles were presented with history of diarrhea, dehydration, weight loss and passage of undigested food in the faeces. Direct faecal examination and Giemsa staining revealed the presence of cysts of Nyctotherus sp. Treatment was initiated with antiprotozoal drug, Metronidazole at the rate of 25 mg/kg body weight per-oral consecutively for 5 days, and Vimeral suspension at rate of 10-20 mg/kg per-oral for 8 days. After 10 days of intensive care and management with antiprotozoal drug, multivitamin under proper sanitary measures, the case showed gradual improvement in condition. During medication, the fecal samples were checked on every alternative day for the presence of ciliated trophozoites of Nyctotherus sp. After 4th day, there was no trophozoites observed by both the techniques examined and the animal was quite healthy and active and started taking food. Improvement was noticed with resumption of normal appetite. Incessant patient monitoring and initiation of suitable need based therapeutic strategy is most important in management and control of Nyctotherus sp. infection.

Isolation and characterization of Toxoplasma gondii from small ruminants (sheep and goats) in Chennai City, South India.

The present study aimed for the isolation and genotyping of Toxoplasma gondii from small ruminants (sheep and goats). 14 out of 193 tissue samples (either brain and heart) tested positive by MDAT for anti-T. gondii antibodies, were selected and bioassayed, which resulted 4 samples positive for T. gondii after 40 days of post inoculation. Four samples consisting of 3 numbers of sheep and 1 number of goat tissues out of 14 samples detected by B1 PCR, were genotyped at SAG3 locus by nested polymerase chain reaction-restriction fragment length polymorphism technique (nPCR-RFLP). The results of the present study revealed that the four isolates designated as TgShIn19, TgShIn76, TgShIn77 and TgGtIn27 were circulating in small ruminants, were belonged to genotypes of type II (TgShIn19) and type III (TgShIn76, TgShIn77 and TgGtIn27) which are in concordance with the previously reported genotypes from other animal species and further this presumptive results indicating that the genotype II and III could be the predominant in different animal species including birds and humans in India.

Detection of Toxoplasma gondii in small ruminants in Chennai using PCR and modified direct agglutination test.

A total of 193 sera samples, along with tissues (lung, heart, and brain) collected from 136 sheep and 57 goats from the Corporation slaughter house, Madras Veterinary College teaching hospital, and private mutton shops from Chennai were tested for Toxoplasma gondii. All the sera samples were tested using modified direct agglutination test. Of the 193 sera samples, 57 (29.5 %) had a minimum titre of 1:20, with 30.14 % (41/136) of sheep and 28.07 % (16/57) of goats being seropositive. Tissue samples from all 193 animals, when subjected to B1 based PCR to detect T. gondii DNA, showed 3.67 and 3.50 % to be positive in sheep and goats, respectively. In the present investigation B1 based PCR detected T. gondii in low numbers, possibly due to limitation of the sample size. The presence of T. gondii in tissues of sheep and goats slaughtered for human consumption in Chennai indicates the role of these food animals as potential sources of infection to human.

Genetic characterization of Toxoplasma gondii from autopsy proven cases of AIDS associated cerebral toxoplasmosis in South India.

Toxoplasma gondii (T.gondii) infection can be devastating in the immunodeficient causing high morbidity and mortality. Due to limited availability of both diagnostic facilities and Highly Active Antiretroviral Therapy (HAART), toxoplasmosis continues to be a significant problem amongst Acquired Immuno Deficiency Syndrome (AIDS) patients in India. While scanty literature is available on T. gondii isolates in animals in India, little is known about the genetic diversity of the parasite in humans. Therefore, the present study investigated the genetic diversity of T. gondii in 25 confirmed cases of cerebral toxoplasmosis developing on the background of human immunodeficiency virus (HIV) infection/AIDS. PCR DNA sequencing was performed at four important genetic loci of T. gondii: BTUB, GRA6, alternative SAG2 (alt SAG2) and SAG3 on DNA from tissues obtained at postmortem. The amplified products from all the cases were successfully sequenced except at one locus for one case. Results of the present study suggest that majority of the patients (22/25; 88%) in South India are infected with strains that are recombinants of type II/III and/or strains representing T. gondii different from the archetypal lineages I, II, and III. In addition, clonal types III, MAS, and MAS variant genotypes were encountered. No clonal type I or II was seen in the present study. In addition, variants were observed at alt SAG2 and SAG3 but BTUB and GRA6 were highly conserved. Single nucleotide polymorphisms were observed mainly at two loci which are coding for surface antigens at alt SAG2 and SAG3. In conclusion, the present study reveals genetic diversity in India amongst strains of T. gondii from clinical cases of toxoplasmosis which is in accordance with other recent studies showing a high rate of genetic diversity in this parasite across the globe. There is a need to genotype T. gondii from different forms of toxoplasmosis in humans in India.

DNA Damage in Healthy Individuals and Respiratory Patients after Treating Whole Blood In vitro with the Bulk and Nano Forms of NSAIDs.

Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit COX enzyme activity which affects the inflammatory response. Inflammation is associated with increasing cancer incidence. Pre-clinical and clinical studies have shown that NSAID treatment could cause an anti-tumor effect in cancers. In the present study, blood was taken from healthy individuals (n = 17) and patients with respiratory diseases or lung cancer (n = 36). White blood cells (WBC) were treated with either a micro-suspension, i.e., bulk (B) or nano-suspension (N) of aspirin (ASP) or ibuprofen (IBU) up to 500 µg/ml in the comet assay and up to 125 µg/ml in the micronucleus assay. In this study results were compared against untreated lymphocytes and their corresponding treated groups. The results showed, that NSAIDs in their nano form significantly reduced the DNA damage in WBCs from lung cancer patients in bulk and nano compared to untreated lymphocytes. Also, there was a decrease in the level of DNA damage in the comet assay after treating WBCs from healthy individuals, asthma and COPD groups with aspirin N (ASP N) but not with IBU N. In addition, the number of micronuclei decreased after treatment with NSAIDs in their nano form (ASP N and IBU N) in the healthy as well as in the lung cancer group. However, this was not the case for micronucleus frequency in asthma and COPD patients. These data show that lymphocytes from different groups respond differently to treatment with ASP and IBU as measured by comet assay and micronucleus assay, and that the size of the suspended particles of the drugs affects responses.

Sweet's syndrome with tendinitis.

A 30 years man presented with fever and skin manifestations in the form of multiple tender, erythematous, asymmetric nodules and papules predominantly over the upper arms and back. The nodules were coalescing to form irregular sharply demarcated plaques over the forearms and thighs. There were pustules over the nodules. He also had tendoachilles tendinitis. The total leucocyte count, differential count and skin biopsy confirmed the diagnosis of Sweet's syndrome (acute febrile neutrophilic dermatosis). This patient, a case of Idiopathic Sweet's syndrome with tendoachilles tendinitis responded to oral corticosteroids.

HLA class I and class II association with ankylosing spondylitis in a southern Indian population.

Ankylosing spondylitis (AS) is a chronic inflammatory condition associated with HLA B27. But the association is not absolute. Hence association with other HLA class I antigens and class II alleles was studied in a southern Indian population. Sixty-five patients with primary AS were typed serologically for HLA class I antigens. Age- and sex-matched disease controls (37 with enterogenic reactive arthritis [ReA] and 25 with undifferentiated spondyloarthropathy [UnSpA]) and 124 healthy controls were studied. PCR-based DNA-SSO typing for DQA1 and DQB1 was performed on 20 patients with AS and 38 healthy controls. Twenty-three patients with AS and 39 controls were typed for DRB1 alleles. HLA B27 was positive in 76.9 % of the cases of AS (RR 811), 59.5% of those with ReA (RR 9.3), and 40% of the patients with UnSpA (RR 9.3), while none of the controls were B27 positive. The P value for positive association was highly significant for B27 in all the three groups. B27 positivity was associated with earlier age of onset of disease in all the diseases compared to the B27-negative group. HLA Cw2 was positively associated with AS (P highly significant; OR 52) and ReA (P = 0.0003; OR 14.2). HLA A1 and CW6 were significantly negatively associated only with AS (P = 0.0001 and 0.00004 and OR 0.25 and 0.02, respectively). None of the HLA class II alleles were significantly associated with AS. The apparent association with DRB1*11 (P = 0.03) was lost after Yates correction.