Molecular Mechanisms of Metabolic Reprogramming in Pancreatic Cancer

Pancreatic cancer ranks as third deadliest cancer worldwide in both men and women. The lack of diagnostic methods, chemo-resistance and invasiveness of the disease diminishes the overall survival rate in patients. Dysregulated glucose metabolism and several molecular determinants including oncogenic signaling pathways play pivotal roles in the development, progression and metastasis of pancreatic cancer. The aberrant glucose metabolism is the most prevalent key factor that influences pancreatic tumorigenesis. The cancer cells of pancreas utilize oxygen for the breakdown of glucose to lactate, which is known as Warburg effect. This phenotype of pancreatic cancer encourages proliferation, growth, migration and invasive phenotypes of cancer cells that worsens the disease. The upstream regulatory molecules responsible for deviant activity of glucose transporters during aberrant glucose metabolism was not very well defined. Thus, identification of new molecular determinants that regulate glucose metabolic pathway would provide promising therapeutic outcomes. Our lab has established the role of MUC13 protein in pancreatic cancer development. Another interesting molecule, Protein Kinase D1 has been reported to play essential role in hypoxic metabolism in squamous cell carcinoma, and glucose uptake in cardiac hypertrophy. Also, it has been extensively studied in our research group, and has been suggested by some other groups to have an essential role in pancreatic cancer. However, its role in regulating glucose metabolism in pancreatic cancer remains elusive. In this dissertation, we investigated that MUC13 and PKD1 proteins are involved in abrupt glucose metabolism in pancreatic cancer cells. We have demonstrated that targeted overexpression of these oncogenes upregulates key oncogenic signaling components involved in aberrant glucose metabolism in pancreatic cancer cells. We also investigated a natural agent Steviol that restored glucose metabolism in pancreatic cancer cells via repression of MUC13 and PKD1 expression. MUC13 is a transmembrane glycoprotein which has an elevated expression in pancreatic cancer. Due to the presence of a cytoplasmic domain, it confers kinase activity and regulates many signaling processes. Therefore, we investigated the effect of MUC13 in metabolic reconditioning of pancreatic cancer. As a result, it was observed that presence of MUC13 in pancreatic cancer upregulated glucose uptake and lactate secretion. This in turn led to more aggressive behavior of the tumor, as evidenced by enhanced proliferation, migration and invasive characteristics of the cells as compared to MUC13 null cells. Interestingly, we observed the interaction of MUC13 with Glut-1 protein, which influences glucose uptake in cancer cells. This was confirmed through various assays such as: immunoprecipitation, immunofluorescence, co-capping and proximity ligation assay. The interpretation of inhibitor results envisaged the role of NFҡB pathway during the molecular interactions of MUC13 and Glut-1. To understand the clinical importance of this mechanism, we evaluated this interaction in tumor samples from the patients with pancreatic disease in advanced stages. Our findings fortified the mechanistic role of MUC13 in rewiring of aberrant glucose metabolism. Altogether, we believed that MUC13 has clinical implication as a key molecule that might be responsible for dysregulated glucose metabolism. As discussed earlier about the paucity of associated proteins related to dysregulated glucose metabolism, we further investigated the possible key players with kinase domain for association and activation with other signaling molecules. Protein Kinase D1 (PKD1), a Ser/Thr kinase has been shown to be involved in progression of pancreatic cancer. In this study, we established PKD1 as a novel molecular target and its involvement in regulating aberrant glucose metabolism in pancreatic cancer. We observed that PKD1 was overexpressed in pancreatic cancer tissues compared to normal human tissues. We also found constitutive expression of PKD1 in various pancreatic cancer cells as compared to normal pancreatic ductal epithelial cells. The overexpression of PKD1 in low PKD1 expressing pancreatic cancer cells enhanced tumorigenic characteristics. We observed that specific knockdown of PKD1 inhibited key oncogenic signaling components in pancreatic cancer cells suggesting its oncogenic role in pancreatic cancer. Additionally, during the PKD1 overexpression, cells displayed increased glucose consumption and lactate production. This suggested role of PKD1 in metabolic reprogramming in pancreatic cancer. Also, we observed that PKD1 stimulates the glucose uptake through mTORC1, a component of mTOR signaling pathway. Additionally, the knockdown of PKD1 promoted the chemosensitivity of cells towards gemcitabine and 2DG (2-deoxyglucose). Overall, these results indicated a significant function of PKD1 in rewiring of glucose metabolism in pancreatic cancer. It was mandating to target and counteract the aberrant glucose metabolism along with the proteins identified in our previous results. In this perspective herein, we proposed steviol, a natural sweetener from plant origin to assess its inhibitory action towards dysregulated glucose metabolism and its associated proteins (MUC13 and PKD1). The reason behind steviol selection for this approach includes that it mimics the glucose molecule that facilitates enhanced uptake of drug (steviol) within the cells and on the other side, it provides glucose homeostasis to adjacent normal cells. These features made steviol as dual functional drug by overtaking other natural drugs for promising results. The exposure of steviol inhibited the proliferative (IC50: 10mM) and clonogenic abilities of pancreatic cancer cells. It also suppressed the cell migration and cell invasion capabilities of pancreatic cancer cells. Further, steviol caused the selective inhibition of intracellular glucose intake and lactate accumulation in a concentration dependent manner. The selective inhibition of MUC13 and PKD1 (upstream key proteins of glucose metabolism) that caused the disruption of glucose intake in cancer cells, was observed during steviol treatment. Secondly, steviol interfered with translation-initiation machinery, causing destabilization of the cellular functionality. The impairment of cellular translation process had promoted G1 cell cycle arrest in cells due to the lack of G1-S transition proteins during drug exposure. Collectively, all these events engendered the cellular integrity and directed towards activation of apoptosis. In conclusion, we evaluated the role of MUC13 and PKD1 in aberrant glucose metabolism leading to growth and aggressive metastatic phenotypes of pancreatic cancer. We also demonstrated efficacy of steviol to repress aberrant glucose metabolism via suppression of MUC13 and PKD1 proteins

Management of preterm premature rupture of membrane in a high-risk pregnancy

Preterm premature rupture of membranes (PPROM) usually has a multi factorial aetiology that is often unknown, although the most frequently reported cause is infection, others could be cervical insufficiency, over distention of uterine cavity, previous history of preterm labor, systemic inflammatory disease etc. This case report documented a rare incidence of preterm premature rupture of membranes, in a 35-year-old female with past history of 8 spontaneous abortions and 1 ectopic pregnancy. Patient had an IVF conception, amenorrheic for 7 months and 2 weeks, and came with a complaint of white discharge per vaginum. On per speculum examination, cervical os was found dilated. The patient was managed conservatively with protein and progesterone support along with prophylactic tocolytics and antibiotics, till early signs of chorioamnionitis were noticed. The decision of induction of labour and eventually preterm premature rupture of membranes lead to emergency cesarean section. Challenges faced in managing this high risk pregnancy are discussed, emphasizing the need of close monitoring and tailoring management in similar circumstances.

Assessment of maternal and fetal outcome in trial of labor after cesarean

Background: Nowadays, cesarean sections are increasing consistently. Repeat cesarean sections are performed for a large percentage and associated with a higher rate of surgical complications and Long-term morbidities. The trial of labor after cesarean offers an alternative choice. This study carried out to assess the maternal and fetal outcome and to evaluate various parameters as a predictor of success of TOLAC.Methods: This prospective observational study conducted on 150 pregnant women with one previous LSCS who delivered at Mahatma Gandhi hospital, from January 2017 to July 2018. Patient having a singleton pregnancy, cephalic presentation, adequate pelvis size with spontaneous onset of labor were included. Cases were monitored carefully during the labor. Emergency LSCS was done if any indication appeared.Results: 78% of cases delivered safely by the vaginal birth and 22% of cases had an emergency repeat cesarean section (EmRCS). Favorable Bishop Score, active stage of labor and prior vaginal delivery were associated with higher success rate. One (0.66%) case of uterine scar rupture and 2 (1.33%) cases of scar dehiscence noted. No maternal mortality observed. Perinatal mortality occurred in 2 cases (1.33%).Conclusions: Present study shows that appropriate clinical settings and the properly selected group of patients can make the TOLAC safe and effective

AN EXPEDIENT APPROACH TO TREAT ASTHMA THROUGH NON-STEROIDAL, NATURAL TRANSFEROSOMES AEROSOL SYSTEM

Asthma is the most common respiratory disease, affecting an estimated 262 million people and resulting in 461,000 fatalities in 2019. The treatment is available on the market, but it is quite expensive, and it also has serious adverse effects due to the high concentration of steroids in the medicine. If given effectively, curcumin, formononetin, and matrine’s anti-inflammatory properties can play a significant role in treatment. To improve the chemical stability and therapeutic potential of these active pharmaceutical ingredients (APIs) in the respiratory tract, a transferosomes system was designed, which encapsulates the APIs inside its vesicular structure and delivers them selectively to the inflamed cells. The DPPC layer will allow for efficient penetration, whereas Tween-80 will aid in deformability and lower interfacial tension, resulting in a small Z-average diameter, allowing for efficient penetration between layers of cells. The APIs’ stability at alkaline pH (7.6) is ensured by the nano-vesicular structure, which significantly increases cellular antioxidant activity and ferric reducing antioxidant power values. On the RAW264.7 cell line, the formulation will be tested for anti-inflammatory activity. Nuclear factor kappa B, tumor necrosis factor, interleukin (IL)-1, IL-2, IL-6, IL-8, IL-12, nitric oxide, and cyclooxygenase-2 are all reduced by curcumin, formononetin, and matrine. They also have an inhibitory effect on the MAPK signaling pathway, preventing extracellular signal-regulated kinase, c-Jun N-terminal kinase, and p38 from causing inflammation. This formulation can effectively treat asthma without the use of steroids, has no adverse effects, and is inexpensive

Vaginal Rhabdomyosarcoma in a patient with advanced cervical cancer; a case report and review of literature

Rhabdomyosarcoma is very rare in adults accounting for less than 5% of all soft tissue tumours and less than 1% of all malignancies. Vagina is one of the least common sites for occurrence of Rhabdomyosarcoma in the genital tract. We present a case of a 53-year-old woman who is a follow up case of cervical cancer stage IIIB, managed by radiotherapy and chemotherapy. She was doing well till 5 years of her treatment for cervical cancer when she presented with complaints of pain lower abdomen and discharge per vaginum for 10 days. On examination she was found to have an abdominal mass of 18 weeks size and on local examination there was 4X4 cm fixed mass on lower third of vagina arising from left side. MRI abdomen and pelvis was done. Biopsy from the vaginal mass showed features of Rhabdomyosarcoma. Further follow up of the patient was not possible due to lockdown in view of the pandemic. She was last contacted telephonically on 25th March 2020; she said she was waiting for the lockdown to be lifted so that her further management can take place. This is just one patient; there are many more with other medical conditions all over the world who are losing their lives because of not being able to access medical care due to the present pandemic. New growth in the region of local recurrence in a known malignancy cannot necessarily be the recurrence of the primary tumour. It is important to keep our mind open to other differentials apart from the recurrence of primary malignancy, sometimes it can turn out to be a very rare tumour as we encountered in our case

Gemcitabine Combination Nano Therapies for Pancreatic Cancer

Pancreatic cancer is one of the deadliest causes of cancer-related death in the United States, with a 5-year overall survival rate of 6 to 8%. These statistics suggest that immediate medical attention is needed. Gemcitabine (GEM) is the gold standard first-line single chemotherapy agent for pancreatic cancer but, after a few months, cells develop chemoresistance. Multiple clinical and experimental investigations have demonstrated that a combination or co-administration of other drugs as chemotherapies with GEM lead to superior therapeutic benefits. However, such combination therapies often induce severe systemic toxicities. Thus, developing strategies to deliver a combination of chemotherapeutic agents more securely to patients is needed. Nanoparticle-mediated delivery can offer to load a cocktail of drugs, increase stability and availability, on-demand and tumor-specific delivery while minimizing chemotherapy-associated adverse effects. This review discusses the available drugs being co-administered with GEM and the limitations associated during the process of co-administration. This review also helps in providing knowledge of the significant number of delivery platforms being used to overcome problems related to gemcitabine-based co-delivery of other chemotherapeutic drugs, thereby focusing on how nanocarriers have been fabricated, considering the modes of action, targeting receptors, pharmacology of chemo drugs incorporated with GEM, and the differences in the physiological environment where the targeting is to be done. This review also documents the focus on novel mucin-targeted nanotechnology which is under development for pancreatic cancer therapy

Estimation of Pesticide Residues in Fruits, Vegetables and Water

Nowadays, pesticide use is on increase day by day in every field of life like agriculture, household and in industry. Most of the pesticides are not biodegradable and those which are degradable cannot degrade completely. Hence, they reach to our fruits, vegetables, grains and water bodies. Even after washing the pesticide residues left in our foodstuffs. Since last decade, it has been observed that analysed pesticide residues were either BDL or < MRL in the majority of samples. In this study vegetable, fruits and grains are an important food item and proper care should be taken to use the very safe pesticide fort avoiding potential risks to human. It is, therefore, suggested that the food item collected from in and around Lucknow City, India is comparatively safe from pesticide residues. A periodical monitoring of pesticide residues in other food commodities is the recent need for the consumers as well as authorities of food quality control

Estimation of Pesticide Residues in Fruits, Vegetables and Water

Nowadays, pesticide use is on increase day by day in every field of life like agriculture, household and in industry. Most of the pesticides are not biodegradable and those which are degradable cannot degrade completely. Hence, they reach to our fruits, vegetables, grains and water bodies. Even after washing the pesticide residues left in our foodstuffs. Since last decade, it has been observed that analysed pesticide residues were either BDL or < MRL in the majority of samples. In this study vegetable, fruits and grains are an important food item and proper care should be taken to use the very safe pesticide fort avoiding potential risks to human. It is, therefore, suggested that the food item collected from in and around Lucknow City, India is comparatively safe from pesticide residues. A periodical monitoring of pesticide residues in other food commodities is the recent need for the consumers as well as authorities of food quality control

Estimation of pesticide residues in fruits, vegetables and water

Nowadays, pesticide use is on increase day by day in every field of life like agriculture, household and in industry. Most of the pesticides are not biodegradable and those which are degradable cannot degrade completely. Hence, they reach to our fruits, vegetables, grains and water bodies. Even after washing the pesticide residues left in our foodstuffs. Since last decade, it has been observed that analysed pesticide residues were either BDL or < MRL in the majority of samples. In this study vegetable, fruits and grains are an important food item and proper care should be taken to use the very safe pesticide fort avoiding potential risks to human. It is, therefore, suggested that the food item collected from in and around Lucknow City, India is comparatively safe from pesticide residues. A periodical monitoring of pesticide residues in other food commodities is the recent need for the consumers as well as authorities of food quality control