Insights into the Emerging Therapeutic Targets of Triple-negative Breast Cancer.

Triple-negative Breast Cancer (TNBC), the most aggressive breast cancer subtype, is characterized by the non-appearance of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER2). Clinically, TNBC is marked by its low survival rate, poor therapeutic outcomes, high aggressiveness, and lack of targeted therapies. Over the past few decades, many clinical trials have been ongoing for targeted therapies in TNBC. Although some classes, such as Poly (ADP Ribose) Polymerase (PARP) inhibitors and immunotherapies, have shown positive therapeutic outcomes, however, clinical effects are not much satisfiable. Moreover, the development of drug resistance is the major pattern observed in many targeted monotherapies. The heterogeneity of TNBC might be the cause for limited clinical benefits. Hence,, there is a need for the potential identification of new therapeutic targets to address the above limitations. In this context, some novel targets that can address the above-mentioned concerns are emerging in the era of TNBC therapy, which include Hypoxia Inducible Factor (HIF-1α), Matrix Metalloproteinase 9 (MMP-9), Tumour Necrosis Factor-α (TNF-α), ß-Adrenergic Receptor (ß-AR), Voltage Gated Sodium Channels (VGSCs), and Cell Cycle Regulators. Currently, we summarize the ongoing clinical trials and discuss the novel therapeutic targets in the management of TNBC.

Novel Antitubercular Agents: Design, Synthesis, Molecular Dynamic and Biological Studies of Pyrazole - 1,2,4-Triazole Conjugates.

Mycobacterium tuberculosis (Mtb) has numerous cell wall and non-cell wall mediated receptors for drug action, of which cell wall mediated targets were found to be more promising because of their pivotal role in bacterial protection and survival. Herein, we reported the design and synthesis of a series of pyrazole-linked triazoles based on the reported structural features of promising drug candidates that target DprE1 receptors through a Structure-based drug design (SBDD) approach (6a-6j and 7a-7j). The synthesized compounds were evaluated for their in-vitro antitubercular activity against virulent strains of Mtb H37Rv. In-silico studies revealed that most compounds exhibit binding interactions with crucial amino acids like Lys418, Tyr314, Tyr60, and Asp386 at DprE1. Furthermore, the protein-ligand (7j) shows appreciable stability compared to innate protein in a 100 ns molecular dynamic simulation study. In-vitro MAB assay revealed that 14 compounds exhibit significant antitubercular activity with minimum inhibitory concentration (MIC) of the 3.15-4.87 µM of the 20 compounds tested. An in-vitro cytotoxicity study on normal cell lines (MCF10) revealed safe compounds (IC50 values:341.85 to 726.08 µM). Hence, the present study opens the development of new pyrazole-linked triazoles as probable DprE1 inhibitors.

The case for the use of telehealth for abortion in India.

India has the world's fastest growing outbreak of COVID-19. With limited mobility, increased reports of intimate partner violence, changes in living patterns of migrants, delays in accessing contraception and safe abortion care, and potential changes to decisions about parenting, there may be an increased need for abortion services in India due to the pandemic. The use of technology for providing abortion information and services has been well documented in global literature. The safety of abortion provision using telehealth has been established in several contexts including the United States and Australia. The importance of hotlines and other support systems that use technology to provide information and support to clients through their abortion is also highlighted in the literature. Several countries, such as the United Kingdom, France, New Zealand, and Pakistan are now allowing the use of technology for abortion/post-abortion care in light of the pandemic; however, India's telemedicine guidelines do not include abortion. In a country where the majority of abortions take place outside the health system, allowing the use of telemedicine for abortion can help bring legality to users, and expand access to those facing additional barriers in accessing the care they deserve. We outline models for telemedicine provision of abortion in India and discuss the regulatory changes required to make telehealth for abortion a reality in India.

Substituted Benzamides from Anti-inflammatory and p38 Kinase Inhibitors to Antitubercular Activity: Design, Synthesis and Screening.

BACKGROUND: Twenty one amide compounds possessing phenoxy/benzyloxy/pyridinyl groups have been synthesized by benzoylation of respective amines in presence of base with moderate to encouraging yields. Upon confirmation of structure, compounds were subjected for p38 kinase inhibitory, anti-inflammatory, antimicrobial and antitubercular activities. METHOD: Anti-inflammatory activity was determined using carrageenan induced rat paw edema model while p38 kinase inhibitory activity was studied using ELISA method and serial dilution method was employed to determine MICs. Two compounds 4g and 4n showed over 30% p38 kinase inhibitory activity at 10 µM and best anti-inflammatory activity was found for compounds 4g, 4i, 4n and 4o which exhibited to reduce paw edema over 70%. Compound 4b was observed to be the most potent against gram +ve organisms with MIC value of 1.6 µG/mL and compound 4u displayed potent antibacterial activity against gram negative organisms. CONCLUSION: Most encouraging antitubercular activity was noticed for compounds 4u, 4r and 4k with 6.25, 12.5 and 12.5 µG/mL Further, in order to know the binding site interactions, a docking simulations of compounds was performed. These preliminary results will certainly show fruitful directions to improve the activities of compounds.

Stepwise case selection using Guy's stone score reduces complications during percutaneous nephrolithotomy training.

INTRODUCTION: Traditional percutaneous nephrolithotomy (PCNL) training involved subjective award of cases to the trainee. We restructured this according to the Guy's stone score (GSS) such that each trainee stepwise completed 25 cases of each grade before progressing. This study compares the outcomes of training with traditional versus stepwise approach. METHODS: Four hundred consecutive cases equally distributed for two trainees in each group were compared in terms of complications (Clavien-Dindo), stone free rate (SFR), operative and fluoroscopy time. External comparison was also done against a benchmark surgeon. Multivariable regression model was created to compare SFR and complications while adjusting for comorbidity, Amplatz size, access tract location, number of punctures, body mass index, stone complexity, and training approach. RESULTS: The distribution of cases in terms of calculus complexity was similar. Overall, in comparison to traditional training, stepwise training had significantly shorter median operative time (100 vs. 120 min, P < 0.05), fluoroscopy time (136 vs. 150 min, P < 0.05) and fewer overall (29.5% vs. 43.5%, P < 0.005) as well as major complications (3% vs. 8.5%, P - 0.029), though initial SFR was higher but not statistically significant (77% vs. 71.5%). On multivariable analyses, stepwise training was independently associated with lower complications (odds ratio 0.46 [0.20-0.74], P - 0.0013) along with GSS grade, number of punctures, and Amplatz size. Stepwise training had similar fluoroscopy time, major complications and final clearance rate compared to expert surgeon. CONCLUSIONS: PCNL has a learning curve specific for each grade of calculus complexity and stepwise training protocol improves outcomes.

Association between QTd, Tp-e/QT Ratio and In-hospital Prognosis in Thrombolysed Acute ST-elevation Myocardial Elevation (STEMI) Patients.

BACKGROUND: Both QTd and Tp-e/QT ratio have been linked to increased risk for arrhythmia and mortality. But Significance of QTd in STEMI Patients is not documented in all studies and Tp-e/QT ratio is a novel index which is understudied in these patients. Therefore, the present study is aimed to determine the short term, in-hospital prognostic value of QTd and Tp-e/QT ratio in thrombolysed STEMI patients. METHODS: This is a prospective, observational study that includes 321 patients. Relevant clinical data is collected. QTd and Tp-e/QT ratio (tangent method) is calculated from "at admission ECG" just before thrombolysis. Multivariate logistic regression analysis was done to determine the predictors of in-hospital outcomes. A p-value of >0.05 is considered statistically significant. RESULTS: The mean age of study population was 56.72 ± 11.36 with males:females ratio of 2.73:1. Mean value of QTd and Tp-e/QT ratio were 80.29 ±10.2 ms and 0.28 ± 0.05 respectively. The QTd and Tp-e/QT ratio are found to be independent predictors of in MACE, in addition to absence of beta-blocker therapy at admission, AWMI. Tp-e/QT ratio is independent predictor of in-hospital mortality in addition to reduced LVEF and AWMI. Analysis of the ROC curve demonstrated that the optimal cut-off value for in-hospital outcomes was a Tp-e/QT ratio of ≥0.30. CONCLUSION: Both QTd and Tp-e/QT ratio may serve as a prognostic predictors of in hospital MACE independently but only Tp-e/QT ratio predicts patients with in-hospital all cause mortality in thrombolysed STEMI patients.

Cytogenetic profiling by FISH microscopy and comparison with light microscopy and complete blood count findings in myelodysplastic syndrome.

Karyotype, bone marrow blast percentage and cytopenia influence the prognosis of myelodysplastic syndrome. We studied the abnormalities detected by fluorescence in situ hybridization (FISH) in myelodysplastic syndrome and associated haematological profile with abnormalities detected by FISH. Complete blood counts, peripheral blood and bone marrow of patients were evaluated for cytopenia, dysplasia and blasts. FISH probes were used to detect del(5q), gain of chromosome 8, de (7q/-7) and del(20 q). Multiple regression analysis was used to study the association of FISH abnormalities, age and sex with haematological profile. Mc Nemar's test studied the relationship between FISH abnormalities and dysplastic features in bone marrow. Cytogenetic abnormalities were detected by FISH in 25.7% of patients. Del(20 q) was seen in 14.2% of patients. FISH was able to predict changes in peripheral blood blast count by 80% (p ˂ 0.0001). Cytogenetic abnormalities were not seen in 74.2% of patients. Groups with FISH abnormalities have a different haematological profile, and these abnormalities have a significant effect on blast percentage.

Alfa-fetoprotein secreting ovarian sex cord-stromal tumor.

Ovarian sex cord-stromal tumors are relatively infrequent neoplasms that account for approximately 8% of all primary ovarian tumors. They are a heterogeneous group of neoplasms composed of cells derived from gonadal sex cords (granulosa and Sertoli cells), specialized gonadal stroma (theca and Leydig cells), and fibroblasts. They may show androgenic or estrogenic manifestations. We report such a tumor associated with markedly raised serum alpha-fetoprotein (AFP) levels in a young female presenting with a mass and defeminising symptoms. Serum AFP levels returned to normal on removal of tumor.

An in vitro comparison of penetration depth of two root canal sealers: An SEM study.

AIM: The aim of this study was to examine in vitro penetration depth of two resin-based sealers (AH plus and Resino Seal) and Zinc Oxide Eugenol sealer into the dentinal tubules after removing smear layer by passive ultrasonic irrigation. MATERIALS AND METHODS: Thirty freshly extracted maxillary central incisors were used. The teeth were decoronated, working length established and prepared upto size 40 file. Each root was subjected to passive ultrasonic irrigation with 2.5% sodium hypochlorite. Three different sealers and gutta-percha were used for obturation. Roots were sectioned using hard tissue microtome. These sections were gold sputtered and examined under scanning electron microscope. RESULTS: Statistical analyses of the data were performed using Kruskal-Wallis and Mann-Whitney tests. Statistically significant difference was found between AH Plus sealer and Resino Seal sealer and Zinc Oxide Eugenol sealer. CONCLUSION: The results showed that AH Plus had maximum penetration depth into dentinal tubules.

The impact of altered insulin-like growth factor-I secretion on the neuroendocrine and testicular functions.

There is unequivocal evidence that the biosynthesis and secretion of gonadotropins by the pituitary gland is controlled by the hypothalamic GnRH and the function of the testis is mainly regulated by FSH and LH. However, a number of investigations have suggested the role of other hormones/factors, including insulin-like growth factor-I (IGF-I) in the control of pituitary and gonadal functions. The role of growth hormone (GH) and IGF-I is poorly investigated in humans. In animals with altered IGF-I secretion, the gonadotropin and androgen secretions are affected. Similarly, there is evidence that fertility, the onset of puberty and sexual maturation are affected in some patients with Laron syndrome and in acromegaly. In this minireview, we have presented some data obtained in humans and also included results from several experimental models with altered GH/IGF-I secretion, in the hope that the results from animals will possibly help in understanding the important role of IGF-I in the control of neuroendocrine-testicular function in humans.

Insulin-like growth factor 1 (IGF-1) and aging: controversies and new insights.

Insulin/insulin-like growth factor (IGF) signaling plays a major role in the control of aging and life span in invertebrates. Major extension of life span in growth hormone receptor knock out (GHR-KO) mice that are GH resistant, and subsequently, IGF-I-deficient indicates that similar mechanisms may operate in mammals. This conclusion is supported by association of reduced IGF-I levels and delayed aging in three different GH-deficient mutant mice and in animals subjected to caloric restriction, but is difficult to reconcile with neuroprotective effects of IGF-I and with the suspected role of declining GH levels during aging. We suggest that the role of IGF in the regulation of growth and adult body size is important in mediating the effects of longevity genes on aging and life span. Suspected mechanisms of IGF-I action in aging also include reduced insulin signaling, enhanced sensitivity to insulin, and reduced thermogenesis with diminished oxidative damage of macromolecules being the likely final common pathway of these effects. We suspect that IGF-I is important in evolutionarily conserved mechanisms that link life history, including development, reproduction, and aging with availability of energy resources.

Consequences of growth hormone (GH) overexpression and GH resistance.

Development of transgenic mice overexpressing GH and GHR-KO mice with GH resistance provided novel animal models for study of the somatotropic axis and for identifying GH actions that may be relevant to its current and contemplated use in medicine and agriculture. Studies of phenotypic characteristics of these animals revealed previously unsuspected actions of GH and IGF-I on neuroendocrine functions related to reproduction and to the release of "stress hormones" (glucocorticoids and prolactin). These studies also provided novel and still-disputed evidence for involvement of somatotropic axis in the control of aging and life span and in mediating the actions of longevity genes.

Genes that prolong life: relationships of growth hormone and growth to aging and life span.

Mutant mice with a combined deficiency of growth hormone (GH), prolactin, and thyrotropin, and knockout mice with GH resistance, live longer than their normal siblings. The extension of life span in these animals is very large (up to 65%), reproducible, and not limited to any particular genetic background or husbandry conditions. In addition to demonstrating that genes control aging in mammals, these findings suggest that GH actions, growth, and body size may have important roles in the determination of life span. We describe the key phenotypic characteristics of long-living mutant and knockout mice, with an emphasis on those characteristics that may be related to delayed aging in these animals. We also address the broader topic of the relationship between GH, growth, maturation, body size, and aging, and we attempt to reconcile the well-publicized antiaging action of GH with the evidence that suppression of GH release or action can prolong life.

Testicular endocrine function in GH receptor gene disrupted mice.

The consequences of disruption of GH receptor gene in GH receptor knockout mice on testicular function were evaluated. Adult male GH receptor knockout mice and their normal siblings were divided in to two subgroups and treated with either saline or ovine LH (0.3 microg/g BW) in saline. One hour after saline or LH administration, blood was obtained via heart puncture. Plasma IGF-I, LH, FSH, PRL, androstenedione, and testosterone levels were measured by RIAs. Testicular LH and PRL receptor numbers as well as pituitary LHbeta-subunit and testicular sulfated glycoprotein-2 mRNA levels were measured. Also, testicular morphometric analysis was performed. Unlike in normal, wild-type mice, the circulating IGF-I was undetectable in GH receptor knockout mice. The plasma PRL levels were (P<0.01) higher in GH receptor knockout mice than in their normal siblings. The basal LH secretion was similar in normal and GH receptor knockout mice. However, the circulating FSH levels were lower (P<0.001) in GH receptor gene disrupted mice. Administration of LH resulted in a significant (P<0.001) increase in plasma testosterone levels in both GH receptor knockout and normal mice. However, this testosterone response was attenuated (P < 0.01) in GH receptor knockout mice. Plasma androstenedione responses were similar in both GH receptor knockout and normal mice. Testicular LH and PRL receptor numbers were significantly decreased in GH receptor knockout mice. The results of the morphometric analysis of the testis revealed that the Leydig cell volume per testis was reduced in mice with GH receptor gene disruption. The steady-state of LHbeta-subunit and testicular sulfated glycoprotein-2 mRNA levels were not different in GH receptor knockout mice relative to their normal siblings. The present in vivo study demonstrates that in GH receptor knockout mice, LH action on the testis in terms of testosterone secretion is significantly attenuated and suggests that this is due to a decrease in the number of testicular LH receptors. The reduced number of PRL receptors may contribute to the diminished responsiveness of testicular steroidogenesis to LH by decreased ability to convert androstenedione to testosterone. These changes are most likely due to the absence of circulating IGF-I. These findings provide evidence that systemic IGF-I plays a major modulatory role in testicular endocrine function.

Epithelial defect in prostates of Stat5a-null mice.

The transcription factor Stat5a critically mediates prolactin (PRL)-induced mammary gland development and lactogenesis. PRL also stimulates growth and differentiation of prostate tissue. Specifically, hyperprolactinemia gives rise to prostate hyperplasia, and prostate size is reduced in PRL-deficient mice. We therefore investigated the importance of Stat5a for prostate development and function by examining Stat5a-null mice. The absence of Stat5a in mice was associated with a distinct prostate morphology characterized by an increased prevalence of local disorganization within acinar epithelium of ventral prostates. Affected acini were typically filled with desquamated, granular epithelial cells that had become embedded in dense, coagulated secretory material. These features were reminiscent of acinar cyst formation and degeneration frequently observed in human benign prostate hyperplasia, however, cystic changes in prostate acini of Stat5a-deficient mice were not associated with increased prostate size or morphologic hallmarks of epithelial hyperplasia. Instead, immunohistochemistry of the prostate-specific secretory marker, probasin, suggested that hypersecretory function of the epithelium could underlie local congestion and cyst formation in prostates of Stat5a-null mice. Serum testosterone and PRL levels were normal in Stat5a knockout mice, but prostate PRL receptor expression was reduced as determined by immunohistochemistry. Expression levels or activation states of other PRL signal transduction proteins, including Stat5b, Stat3, Stat1, ERK1, and ERK2 were not altered. The present study offers the first evidence for a direct role of Stat5a in the maintenance of normal tissue architecture and function of the mouse prostate.

Effects of growth hormone overexpression and growth hormone resistance on neuroendocrine and reproductive functions in transgenic and knock-out mice.

Transgenic mice overexpressing growth hormone (GH) exhibit alterations in the function of the hypothalamic-pituitary-gonadal (HPG) axis and the H-P-adrenal axis. Alterations in the turnover of hypothalamic neurotransmitters, in plasma hormone levels, and in regulation of their release are associated with reproductive deficits, particularly in females. Results reported after publication of our minireview on this subject provided evidence that GH-transgenic mice have increased binding of GH to GH binding proteins in plasma, are hyperinsulinemic and insulin resistant, and have major alterations in energy budgets with increased allocation to growth. Reduced life span and fertility of these animals may be related to insufficient allocation of energy to reproduction and maintenance. Growth hormone resistance induced by transgenic expression of an antagonistic bGH analog or by targeted disruption (knock-out, KO) of the GH receptor (GH-R) gene leads to dramatic suppression of plasma levels of insulin-like growth factor-1 (IGF-1), and dwarf phenotype due to reduced growth and increased adiposity. In both models of GH resistance, there are marked reproductive deficits in females, decline of breeding performance of males, and alterations in the function of the HPG axis. In GH-R-KO females, puberty is delayed, and litter size is reduced. Fetal weights are reduced whereas placental weights are increased, and the weight of newborn pups is reduced despite an increase in the length of gestation. In GH-R-KO males, copulatory behavior and fertility are reduced, plasma PRL is elevated, and responses to luteinizing hormone releasing hormone (LHRH) in vivo and to LH in vitro are suppressed. However, reproductive deficits in GH-R-KO mice are very mild when compared to those described previously in IGF-KO animals. Apparently, the amounts of IGF-1 that may be produced locally in the absence of GH stimulation are sufficient for sexual maturation and fertility in both sexes, whereas quantitative deficits in reproductive function reflect absence of GH-dependent IGF-1 production and other consequences of eliminating GH signaling. The reproduction phenotype of the GH-R-KO mice is also mild when compared to dwarf mice that lack GH, prolactin (PRL), and thyroid stimulating hormone (TSH). This is presumably related to the presence of redundant mechanisms in the stimulatory control of the gonads by the pituitary and the ability of animals capable of producing PRL and TSH to compensate partially for the absence of GH signaling.

Somatotroph and lactotroph changes in the adenohypophyses of mice with disrupted insulin-like growth factor I gene.

Pituitary is influenced by circulating and locally produced insulin-like growth factor I (IGF-I). To further elucidate the role of pituitary IGF-I, we compared pituitary morphology of homozygous (IgfI-/-), heterozygous (IgfI+/+), and wild-type (IgfI+/+) fetal and adult mice using light microscopy, immunocytochemistry, in situ hybridization and electron microscopy. In pituitaries of Igf1-/- and Igf1+/- fetal mice (day 18.5) GH RNA signal was decreased. In Igf1-/- adult females, GH cells were significantly diminished in size; GH RNA signal was stronger in Igf1-/- mice compared with IgfI+/+ mice, and the somatotrophs had ultrastructural features of stimulation. The number of PRL cells and PRL hybridization signal were significantly decreased, however plasma PRL levels were elevated in both genders. No changes in other cell types in Igf1-/- mice, and no alterations in Igf1+/- mice were evident. IGF-I treatment for 2 weeks of Igf1-/- mice increased significantly body weights, decreased GH hybridization signal, and had no effect on PRL cells, or PRL plasma levels, whereas in IgfI+/+ mice, PRL RNA signal and PRL plasma levels were markedly increased. In conclusion, IGF-I plays no role in differentiation of pituitary cells, affects the size of somatotrophs in females, and is a stimulator of lactotrophs in both genders.

Suppression of growth hormone does not affect ongoing spermatogenesis in rats.

Recent evidence suggests that growth hormone (GH) may enhance physiologic processes, such as spermatogenesis, in addition to causing classical anabolic effects. We have previously shown that testosterone restores spermatogenesis in rats that were made azoospermic by immunization against gonadotropin-releasing hormone (GnRH). In this study, we investigated whether suppression of GH affects spermatogenesis and the ability of testosterone to restore spermatogenesis following immunization against GnRH and/or growth hormone-releasing hormone (GHRH). Twelve rats were actively immunized against GnRH (anti-GnRH), twelve rats were actively immunized against GHRH (anti-GHRH), six rats were immunized against both GnRH and GHRH (anti-GnRH/GHRH), and six rats served as controls. Two weeks after the second booster, six rats each from the anti-GnRH and anti-GHRH groups as well as the six anti-GnRH/GHRH rats received 24-cm testosterone-filled Silastic implants (T), and the remaining six rats from each of these groups received empty Silastic implants. All rats were euthanized 2 months later. Weights of testes and testicular sperm counts were determined. Serum testosterone, luteinizing hormone (LH), follicle-stimulating hormone (FSH), growth hormone (GH), and insulin-like growth factor-1 (IGF-1) concentrations were determined by radioimmunoassays. Serum GH and IGF-1 were suppressed in anti-GHRH rats. IGF-1 was partially restored by testosterone in anti-GHRH and in anti-GnRH/GHRH rats, but GH was restored to control value in anti-GnRH/GHRH rats. Serum LH and FSH were suppressed in anti-GnRH and anti-GnRH/GHRH rats, but only FSH was partially restored by testosterone. Suppression of GH did not affect maintenance of spermatogenesis. However, because T partially restored GH and IGF-1 levels in anti-GnRH/GHRH rats and because spermatogenesis was found to be restored in these rats, we conclude that GH does not play a role in the maintenance of spermatogenesis in adult rats, but it may be required for the replenishment of germ cells in experimentally induced regressed rat testes.