Advancements, Challenges, and clinical implications of integration of metabolomics technologies in diabetic nephropathy.

BACKGROUND: Diabetic nephropathy (DN), a severe complication of diabetes, involves a range of renal abnormalities driven by metabolic derangements. Metabolomics, revealing dynamic metabolic shifts in diseases like DN and offering insights into personalized treatment strategies, emerges as a promising tool for improved diagnostics and therapies. METHODS: We conducted an extensive literature review to examine how metabolomics contributes to the study of DN and the challenges associated with its implementation in clinical practice. We identified and assessed relevant studies that utilized metabolomics methods, including nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) to assess their efficacy in diagnosing DN. RESULTS: Metabolomics unveils key pathways in DN progression, highlighting glucose metabolism, dyslipidemia, and mitochondrial dysfunction. Biomarkers like glycated albumin and free fatty acids offer insights into DN nuances, guiding potential treatments. Metabolomics detects small-molecule metabolites, revealing disease-specific patterns for personalized care. CONCLUSION: Metabolomics offers valuable insights into the molecular mechanisms underlying DN progression and holds promise for personalized medicine approaches. Further research in this field is warranted to elucidate additional metabolic pathways and identify novel biomarkers for early detection and targeted therapeutic interventions in DN.

CRISPR/Cas-New Molecular Scissors in Diagnostics and Therapeutics of COVID-19.

The current pandemic of COVID-19, with its climbing number of cases and deaths, has us searching for tools for rapid, reliable, and affordable methods of detection on one hand, and novel, improved therapeutic strategies on the other. The currently employed RT-PCR method, despite its all-encompassing utility, has its shortcomings. Newer diagnostic tools, based on the Clustered Regularly Interspaced Short Palindromic Repeats/Cas(CRISPR-Cas) system, with its better diagnostic accuracy measures, have come up to fill that void. These assay platforms are expected to slowly take up the place of COVID-19 diagnostics. Further, the current therapeutic options focus mainly on counteracting the viral proteins and components and their entry into host cells. The CRISPR-based system, especially through the RNA-guided Cas13 approach, can identify the genomic characteristics of SARS-CoV-2 and provide a novel inhibition strategy for coronaviruses. In this mini-review, we have discussed the available and upcoming CRISPR-based diagnostic assays and the potential of the CRISPR/Cas system as a therapeutic or prevention strategy in COVID-19. CRISPR-Cas system shows promise in both diagnostics as well as therapeutics and may as well change the face of molecular diagnosis and precision medicine.

Are Medical Students from Across the World Aware of Cervical Cancer, HPV Infection and Vaccination? A Cross-Sectional Comparative Study.

This study aims to understand and compare the level of knowledge, the attitude and the awareness of cervical cancer, HPV infection and vaccination among medical students from across the globe. It was conducted by sharing a questionnaire on medical students' Facebook groups. We successfully analysed 736 responses, of which 74.3% were females and 25.7% males. Their mean age was 23.14. As regards to the respondents' knowledge of the risk factors for cervical cancer, 28.8% of the participants identified HPV, 23.5% chose "Having many sexual partners", 15.4% identified "Starting the sexual life at a young age", 14.7% chose HIV and 13.3% answered smoking. Regarding the knowledge of the Pap test, 92.4% of the students stated that they knew what it is, although 98.5% of these respondents chose the correct answer. Concerning the knowledge of early warning signs of cervical cancer, 34.4% recognised vaginal bleeding. According to our research, American, Eastern and Western European students had more knowledge regarding the risk factors, early signs of cervical cancer and Pap test than African and Asian students. A total of 50% of the sexually active females had gone at least once to be screened by the means of a Pap test. A total of 39.8% of the participants stated that they had received the HPV vaccine. The findings of our research highlight the need for additional education measures to improve knowledge and awareness regarding HPV infection, especially among medical students, since they will be future healthcare providers.

CO2 utilization: Turning greenhouse gas into fuels and valuable products.

This study critically reviews the recent developments and future opportunities pertinent to the conversion of CO2 as a potent greenhouse gas (GHG) to fuels and valuable products. CO2 emissions have reached an alarming level of around 410 ppm and have become the primary driver of global warming and climate change leading to devastating events such as droughts, hurricanes, torrential rains, floods, tornados and wildfires across the world. These events are responsible for thousands of deaths and have adversely affected the economic development of many countries, loss of billions of dollars, across the globe. One of the promising choices to tackle this issue is carbon sequestration by pre- and post-combustion processes and oxyfuel combustion. The captured CO2 can be converted into fuels and valuable products, including methanol, dimethyl ether (DME), and methane (CH4). The efficient use of the sequestered CO2 for the desalinization might be critical in overcoming water scarcity and energy issues in developing countries. Using the sequestered CO2 to produce algae in combination with wastewater, and producing biofuels is among the promising strategies. Many methods, like direct combustion, fermentation, transesterification, pyrolysis, anaerobic digestion (AD), and gasification, can be used for the conversion of algae into biofuel. Direct air capturing (DAC) is another productive technique for absorbing CO2 from the atmosphere and converting it into various useful energy resources like CH4. These methods can effectively tackle the issues of climate change, water security, and energy crises. However, future research is required to make these conversion methods cost-effective and commercially applicable.

Glucose Transporter 1 Overexpression in Oral Squamous Cell Carcinoma.

OBJECTIVE: To determine association of immunohistochemical over expression of GLUT 1 (Glucose transporter 1) in oral squamous cell carcinoma (SCC) with histopathological grade and smoking. STUDY DESIGN: Descriptive cross-sectional study. PLACE AND DURATION OF STUDY: Pathology Department, King Edward Medical University (KEMU), Lahore, from January 2018 to July 2018. METHODOLOGY: Paraffin blocks of diagnosed cases of oral SCC presenting at Pathology Department, KEMU, were selected for immunostain GLUT 1. Tumor was graded by WHO 2010 grading. Immunoreactive score (IRS) was calculated for GLUT 1, by multiplying proportion and intensity score of stain. Data was analysed by SPSS 21. Chi-square test was used to measure correlation between GLUT 1 staining, smoking and grade of tumor. P<0.005 was taken as significant. RESULTS: A total of 60 biopsies were included in the study. GLUT 1 was positive in 52 (86.6%) and negative in 8 (13.3%) biopsies. When differentiation of tumor was compared with GLUT 1 positivity with the help of Chi-square test p<0.001 and 95% CI, out of 52 positive biopsies 32 (61.5%) were well, 18 (34.6%) were moderately and 2 (3.8%) were poorly differentiated. GLUT 1 was positive in 43 (82.7%) and only 9 (17.31%) of non-smokers. GLUT 1 was negative in 7 (87.5%) smokers and positive in only 1 (12.5%) of smokers. CONCLUSION: GLUT 1 is positive diffusely in oral SCC with higher expression in lower grades of tumor. As the tumor loses squamous differentiation, it also loses GLUT 1 receptors and thus expression. Smoking has no significant relation with tumor differentiation or GLUT 1 expression.

CO2 capture and storage: A way forward for sustainable environment.

The quest for a sustainable environment and combating global warming, carbon capture, and storage (CCS) has become the primary resort. A complete shift from non-renewable resources to renewable resources is currently impossible due to its major share in energy generation; making CCS an imperative need of the time. This study, therefore, aims to examine the reckoning of carbon dioxide (CO2), measurement methods, and its efficient capture and storage technologies with an ambition to combat global warming and achieve environmental sustainability. Conventionally, physical, geological and biological proxies are used to measure CO2. The recent methods for CO2 analyses are spectrometry, electrochemical gas sensors, and gas chromatography. Various procedures such as pre, post, and oxyfuel combustion, and use of algae, biochar, and charcoal are the promising ways for CO2 sequestration. However, the efficient implementation of CCS lies in the application of nanotechnology that, in the future, could provide a better condition for the environment and economic outlooks. The captured carbon can be stored in the earth crust for trillions of years, but its leakage during storage can raise many issues including its emissions in the atmosphere and soil acidification. Therefore, global and collective efforts are required to explore, optimize and implement new techniques for CCS to achieve high environmental sustainability and combat the issues of global warming.

Prognostic Value of p53 Expression Intensity in Urothelial Cancers.

OBJECTIVE: To determine association of immunohistochemical expression intensity of p53 with grade and stage of urothelial cancers. STUDY DESIGN: Descriptive cross-sectional analytical study. PLACE AND DURATION OF STUDY: Pathology Department, King Edward Medical University, Lahore, from January to December 2016. METHODOLOGY: Data of transurethral resection/radical cystesctomy urinary bladder biopsies was collected. Clinical, radiological and cystoscopic findings of patients were noted from patients' charts in the Urology Ward. Biopsies were graded histologically according to WHO 2004 grading system. TNM system was used for pathological staging. On selected slides, immunoshistochemistry for p53 was applied. Nuclear immunoreactivity was considered positive if present in >10% of tumor cells and negative if <10% of tumor cells. Intensity was considered weak (less than 15% cells) and strong (more than 15% cells). Data was analyzed by SPSS version 21. Linear-by-linear association was calculated between p53 expression and stage of urothelial tumors, Chi-Square test was used to see association between grade and intensity of p53. Qualitative variables, like grade and stage of carcinoma along with p53 expression, were calculated in terms of frequencies and percentages. P ≤ 0.05 was taken as significant. RESULTS: Out of the 70 patients, 61 (87%) were males and 9 (13%) females. Out of 25 low grade lesions, 4 (16%) cases were p53 positive; and out of 45 high grade lesions, 41 (91%) cases were p53 positive. There was 33% (2/6 cases) positivity in Tis, 55% (16/29 cases) in T1, 72% in T2 (21/29), and 100% in T3a (5/5 cases) and T3b (1/1 case). Strong intensity of p53 staining was noted to be 5.4% (n=25) of low grade and 94.6% (n=45) of high grade tumors. CONCLUSION: p53 expression was greater and more frequently strong in higher grade and stage of urothelial carcinoma. It can be used as a prognostic marker in predicting higher grade and stage of bladder cancer.

Medical student clerkship performance and career selection after a junior medical student surgical mentorship program.

BACKGROUND: The impact of early medical school mentorship in students' clerkships performance and career selection is unknown. METHODS: We administered Introduction to Surgery, a resident-directed, semester-long, preclinical elective to junior medical students who answered a Likert-type survey after residency application. Elective participants (EPs) were compared with nonparticipant applicants (EAs), medical school class (MS), and national match outcomes (USA). RESULTS: All 18 EPs (7 M1's, 11 M2's) completed the elective and survey. EP reported more confidence and improved surgical skills, especially attributed to resident mentorship (F(13,237) = 2.3, P = 8*10(-3)). EP "honored" the clerkship more than MS (P = .05); 55.6% of EP, 37.5% of EA, and 27.7% of MS chose surgical fields, yielding a relative risk of 2.0 for EP vs MS (95% confidence interval: 1.3 to 3.2, P = 4*10(-3)). EP "strongly agree" with future mentorship programs (4.6/5), and 1 EP reported the course to be the "main reason" for applying to general surgery. CONCLUSIONS: Introduction to Surgery provides a model for a multifaceted junior medical student mentorship program, which has the potential to retain interested students for surgical career selection.

Deformation luminescence produced during application and release of pressure on to gamma-irradiated CaF2:RE crystals.

Calcium fluoride CaF2 is an interesting host lattice for rare earth (RE) activators. CaF2 crystals doped with different concentrations of Dy, Ce, Er and Gd have been grown by the Bridgman technique and their deformation luminescence (DL) induced by room temperature gamma irradiation has been recorded. When a uniaxial pressure is applied on to gamma-irradiated CaF2:RE crystals, initially the DL intensity increases with time, attains a maximum value and then it decreases with time. Although the DL intensity produced during the release of pressure is less, its rise and decay behaviours are similar to that obtained during the application of pressure. The DL intensity depends on dopant, concentration of dopant, irradiation doses and mass of the load or applied pressure. It is suggested that the moving dislocation produced during deformation of crystals capture holes from hole trapped centres (like perturbed Vk centre) and the subsequent radiative recombination of the dislocation holes with electrons give rise to DL.

Correlation between thermoluminescence and mechanoluminescence of gamma-irradiated Dy activated potassium and magnesium mixed sulphate.

Thermoluminescene (TL) and mechanoluminescence (ML) properties of gamma-irradiated Dy activated potassium and magnesium mixed sulphate have been studied. (K2Mg2:Dy) (SO4)3 samples having different concentrations of Dy were prepared by solid-state diffusion method. Two distinct peaks, the first approximately 130 degrees C and the second approximately 273 degrees C, are observed in the TL glow curve. It is also observed that TL intensities of both peaks decrease when TL glow curves were recorded after deforming the irradiated samples. Only one peak is observed in the ML intensity vs. time curve and the ML intensity decreases markedly with the post-irradiation annealing (to remove 130 degrees C TL peak) of the sample. Both ML and TL intensities have been observed optimum for 1 mol% of Dy in the mixed sulphate system. It is suggested that the recombination of electrons with the free radicals (anion radicals produced by gamma irradiation) released from the traps during the thermal or mechanical excitation is responsible for luminescence in this system.

Indoor radon concentration in the rural dwellings of Chhattisgarh state (India).

Inhalation of radon and its daughter products is the major contributor to the total exposure of the population to natural radiation. An indoor radon survey has been carried out in the state of Chhattisgarh (80.26 degrees N to 84.41 degrees N and 17.8 degrees E to 24.1 degrees E), India under the national coordinated radon project of the Department of Atomic Energy. In the frame of this project indoor radon concentration has been measured in 105 dwellings situated in different villages of Chhattisgarh state. Houses were selected for measurements to cover the most common type of houses generally existing in the rural areas. Measurements have been done on quarterly integrating cycle for one full year in each dwelling using radon cup dosemeter employing LR-115, type-II (pelliculable), solid-state nuclear track detectors. The gamma radiation level was also checked in each dwelling using a gamma survey meter. It was found that the annual average indoor radon concentration in these dwellings varies from 9.91 to 87.84 Bq m(-3) with overall mean value of 26.48 Bq m(-3). Gamma level in these rural dwellings varies from 14.84 to 26.56 microR h(-1) with mean value of 18.68 microR h(-1). We observed that the radon concentration is relatively higher in the houses where the floor is bare but relatively lower in those houses where the floor is tiled or cemented.

Mechano and thermoluminescence of gamma-irradiated CaSO4:Dy phosphor.

In order to have a better idea of the interaction of the defect centres produced by y-irradiation with dislocation in the processes of deformation destruction, mechanoluminescence (ML) and thermoluminescence (TL) of gamma-irradiated CaSO4:Dy, these phosphors have been investigated. CaSO4:Dy phosphors were prepared by dissolving CaSO4.2H2O in sulphuric acid and evaporating the excess acid around 300 degrees C. ML was excited impulsively by dropping a load on to the sample. Two peaks have been observed in the ML intensity against time curve. The total light output, i.e. integrated ML intensity, increases with concentration of dopant, strain rate and with irradiation doses. The TL glow curves of CaSO4:Dy phosphors at different concentrations of dopant and irradiation doses were also recorded. Studies of the influence of post-irradiation annealing on the ML of CaSO4:Dy show that with the removal of the TL dosemetric peak (approximately 210 degrees C) the ML intensity decreases markedly. A spectroscopic study of ML and TL has also been carried out to elucidate the mechanism of ML. Correlation between ML and TL has also been found.

Long-term therapy of chronic hepatitis B with lamivudine.

Lamivudine therapy induces improvements in chronic hepatitis B in a high proportion of patients, but prolonged therapy is limited by the development of viral resistance. We analyzed clinical responses and virologic resistance in 27 patients treated continuously with lamivudine for 2 to 4 years. Serum transaminases, hepatitis B virus (HBV) DNA by both branched DNA (bDNA) signal amplification and quantitative polymerase chain reaction were monitored at 4- to 8-week intervals. Virologic resistance to lamivudine was confirmed by the presence of mutations in the YMDD motif of the polymerase gene by restriction fragment-length polymorphism analysis. Serum HBV-DNA levels decreased rapidly in all treated patients, falling by 4 to 5 logs within 1 year. Transaminase levels also decreased and were normal in 70% of patients at 1 year, at which point liver histology had improved in 81% of patients. Viral resistance began to emerge after 8 months of therapy, eventually developing in 14 patients, including 76% of hepatitis B e antigen (HBeAg)-positive patients but only 10% of HBeAg-negative patients. Lamivudine withdrawal led to reappearance of wild-type HBV species, but retreatment led to more rapid reappearance of the mutant virus. Clinical, serum biochemical, and histologic improvements were maintained in the 13 patients who did not develop resistance. Thus, long-term therapy with lamivudine resulted in maintained improvements in virologic, biochemical, and histologic features of disease in most patients with HBeAg-negative chronic hepatitis B and in the subgroup of HBeAg-positive patients with high serum transaminase levels. A high rate of resistance limited efficacy, particularly in patients who remained HBeAg positive on therapy.

Applications of comparative genomic hybridisation in constitutional chromosome studies.

G band cytogenetic analysis often leads to the discovery of unbalanced karyotypes that require further characterisation by molecular cytogenetic studies. In particular, G band analysis usually does not show the chromosomal origin of small marker chromosomes or of a small amount of extra material detected on otherwise normal chromosomes. Comparative genomic hybridisation (CGH) is one of several molecular approaches that can be applied to ascertain the origin of extra chromosomal material. CGH is also capable of detecting loss of material and thus is also applicable to confirming or further characterising subtle deletions. We have used comparative genomic hybridisation to analyse 19 constitutional chromosome abnormalities detected by G band analysis, including seven deletions, five supernumerary marker chromosomes, two interstitial duplications, and five chromosomes presenting with abnormal terminal banding patterns. CGH was successful in elucidating the origin of extra chromosomal material in 10 out of 11 non-mosaic cases, and permitted further characterisation of all of the deletions that could be detected by GTG banding. CGH appears to be a useful adjunct tool for either confirming deletions or defining their breakpoints and for determining the origin of extra chromosomal material, even in cases where abnormalities are judged to be subtle. We discuss internal quality control measures, such as the mismatching of test and reference DNA in order to assess the quality of the competitive hybridisation effect on the X chromosome.

Cytogenetic abnormalities in the leukemic phase of non-Hodgkin lymphoma.

We have carried out chromosome analysis in a series of 16 non-Hodgkin Lymphoma (NHL) cases in leukemic phase. The diagnoses in these patients based on histology and immunologic markers were as follows: follicular lymphoma (FL), 3 cases; mantle cell lymphoma (Mc), 4 cases; lymphoplasmacytic lymphoma (LPL), 8 cases, and large cell lymphoma, 1 case. We have shown that the t(14;18), t(11;14), and trisomy 12 retained their subtype association with FL, Mc, and LPL, respectively, as in their nonleukemic counterparts with one case of FL showing t(1;19)(q23;p13). Among the four LPL cases without trisomy 12, one case each showed t(12;14)(q13;q32), trisomy 14, t(1;3)(p34;q21), and del(3)(q21). The t(1;19) and t(12;14) may represent rare events in FL and LPL, respectively, and may be uniquely associated with the leukemic phase. The breakpoint 14q32 was the most common single breakpoint involved, sometimes involving both chromosome 14 homologues depicting its association with primary and secondary genetic events in the disease progression. In addition to the main abnormalities, we have shown additional complex abnormalities in 14 of 16 cases. Among these, chromosome 3 was the most commonly involved, affecting the short or long arm or the whole chromosome; 5 of the 16 cases involved breakpoint 3q21. The high incidence of additional abnormalities in these NHL in leukemic phase suggest an association with the development of leukemia and progression of the disease.

Silent cardiac ischemia in cervical spinal cord injury: case study.

The management of ischemic heart disease in patients with chronic spinal cord injury (SCI) will become an increasingly major concern as this population ages. Although silent ischemia has become an important topic in the medical literature, the relationship with cervical SCI has not been adequately explored. A literature search revealed no case reports of documented asymptomatic cardiac ischemia in SCI patients. This is a case report of a 65-year-old patient with chronic C7 incomplete SCI who had multiple risk factors for coronary artery disease and an abnormal electrocardiogram. Despite being completely asymptomatic, the patient was found to have significant myocardial ischemia induced by minimal stress using atrial paced thallium scintigraphy. This finding led to the cancellation of an elective surgical procedure. This case illustrates the importance of suspecting silent myocardial ischemia in cervical SCI patients.

Cytogenetic studies of leukemic recurrence in recipients of bone marrow allografts.

Cytogenetic studies were made in 20 leukemic patients who relapsed after treatment by allogeneic bone marrow transplantation (BMT). Seven of the eight patients in whom no chromosomal abnormalities were detected in leukemic cells before BMT developed clonal abnormalities after BMT, and in two of these patients two independent clones were observed. Most patients in whom clones were detected before BMT showed evidence of clonal evolution after BMT. Nonclonal abnormalities were also observed in clonal cells. These additional abnormalities, both clonal and nonclonal, were attributed to the effects of total body irradiation received by the patient before BMT. There was no evidence of recurrence of leukemia in donor cells among these patients. We concluded that the cells found in leukemic recurrence were derived from the original leukemic clone.

Simultaneous application of immunolabelling and in situ hybridization to detect the origin of B and T lymphocytes in a case of acute lymphocytic leukaemia after bone marrow transplantation.

Previous cytogenetic studies, using selective mitogens, on a patient with B cell acute lymphocytic leukaemia during the 6 years of remission after bone marrow transplantation from an HLA-identical sister indicated persistence of recipient B lymphocytes in the peripheral blood. Such studies are necessarily limited to dividing cells at metaphase, which represent only a small proportion of the total cell population. We have now combined the techniques of immunolabelling and in situ hybridization on the patient's peripheral blood lymphocytes in order to define accurately their individual lineage and gender. A clear difference in the proportion of the persisting recipient lymphocytes was found between B and T lymphocyte lineages.

Cytogenetic studies using Q-band polymorphisms in patients with AML receiving marrow from like-sex donors.

After bone marrow transplantation (BMT), it is important to monitor the bone marrow and lymphoid cell populations of the recipient to document engraftment. When donor and recipient are of unlike sex, the sex chromosomes serve as a useful marker to determine cellular origin. When donor and recipient are of like sex, autosomal heteromorphisms can be used to identify the origin of cells in metaphase. Using Q-banding, we found that 17 of 20 patient/donor pairs (85%) examined showed at least one chromosome heteromorphism that distinguished between recipient and donor cells with certainty. Five of the patients were followed up after BMT in order to document engraftment. Donor metaphases could be detected in the marrow within two weeks of BMT when the graft was successful. Chimaerism was detected in the lymphocyte population even when the graft persisted. In a case of graft failure, donor cells did not persist in the marrow, and the lymphocyte population did not convert to donor type. These studies demonstrate that autosomal heteromorphisms are useful in the study of myeloid and lymphoid chimaeric states after BMT.