Development of Biocompatible Electrospun PHBV-PLLA Polymeric Bilayer Composite Membranes for Skin Tissue Engineering Applications.

Bilayer electrospun fibers aimed to be used for skin tissue engineering applications were fabricated for enhanced cell attachment and proliferation. Different ratios of PHBV-PLLA (70:30, 80:20, and 90:10 w/w) blends were electrospun on previously formed electrospun PHBV membranes to produce their bilayers. The fabricated electrospun membranes were characterized with FTIR, which conformed to the characteristic peaks assigned for both PHBV and PLLA. The surface morphology was evaluated using SEM analysis that showed random fibers with porous morphology. The fiber diameter and pore size were measured in the range of 0.7 ± 0.1 µm and 1.9 ± 0.2 µm, respectively. The tensile properties of the bilayers were determined using an electrodynamic testing system. Bilayers had higher elongation at break (44.45%) compared to the monolayers (28.41%) and improved ultimate tensile strength (7.940 MPa) compared to the PHBV monolayer (2.450 MPa). In vitro cytotoxicity of each of the scaffolds was determined via culturing MC3T3 (pre-osteoblastic cell line) on the membranes. Proliferation was evaluated using the Alamar Blue assay on days 3, 7, and 14, respectively. SEM images of cells cultured on membranes were taken in addition to bright field imaging to visually show cell attachment. Fluorescent nuclear staining performed with DAPI was imaged with an inverted fluorescent microscope. The fabricated bilayer shows high mechanical strength as well as biocompatibility with good cell proliferation and cell attachment, showing potential for skin substitute applications.

Deep Eutectic Solvent Coated Cerium Oxide Nanoparticles Based Polysulfone Membrane to Mitigate Environmental Toxicology.

In this study, ceria nanoparticles (NPs) and deep eutectic solvent (DES) were synthesized, and the ceria-NP's surfaces were modified by DES to form DES-ceria NP filler to develop mixed matrix membranes (MMMs). For the sake of interface engineering, MMMs of 2%, 4%, 6% and 8% filler loadings were fabricated using solution casting technique. The characterizations of SEM, FTIR and TGA of synthesized membranes were performed. SEM represented the surface and cross-sectional morphology of membranes, which indicated that the filler is uniformly dispersed in the polysulfone. FTIR was used to analyze the interaction between the filler and support, which showed there was no reaction between the polymer and DES-ceria NPs as all the peaks were consistent, and TGA provided the variation in the membrane materials with respect to temperature, which categorized all of the membranes as very stable and showed that the trend of stability increases with respect to DES-ceria NPs filler loading. For the evaluation of efficiency of the MMMs, the gas permeation was tested. The permeability of CO2 was improved in comparison with the pristine Polysulfone (PSF) membrane and enhanced selectivities of 35.43 (αCO2/CH4) and 39.3 (αCO2/N2) were found. Hence, the DES-ceria NP-based MMMs proved useful in mitigating CO2 from a gaseous mixture.

Recent Advancements in Metal and Non-Metal Mixed-Doped Carbon Quantum Dots: Synthesis and Emerging Potential Applications.

In nanotechnology, the synthesis of carbon quantum dots (CQDs) by mixed doping with metals and non-metals has emerged as an appealing path of investigation. This review offers comprehensive insights into the synthesis, properties, and emerging applications of mixed-doped CQDs, underlining their potential for revolutionary advancements in chemical sensing, biosensing, bioimaging, and, thereby, contributing to advancements in diagnostics, therapeutics, and the under standing of complex biological processes. This synergistic combination enhances their sensitivity and selectivity towards specific chemical analytes. The resulting CQDs exhibit remarkable fluorescence properties that can be involved in precise chemical sensing applications. These metal-modified CQDs show their ability in the selective and sensitive detection from Hg to Fe and Mn ions. By influencing their exceptional fluorescence properties, they enable precise detection and monitoring of biomolecules, such as uric acid, cholesterol, and many antibiotics. Moreover, when it comes to bioimaging, these doped CQDs show unique behavior towards detecting cell lines. Their ability to emit light across a wide spectrum enables high-resolution imaging with minimal background noise. We uncover their potential in visualizing different cancer cell lines, offering valuable insights into cancer research and diagnostics. In conclusion, the synthesis of mixed-doped CQDs opens the way for revolutionary advancements in chemical sensing, biosensing, and bioimaging. As we investigate deeper into this field, we unlock new possibilities for diagnostics, therapeutics, and understanding complex biological processes.

A novel pH-responsive hydrogel system based on Prunus armeniaca gum and acrylic acid: Preparation and evaluation as a potential candidate for controlled drug delivery.

pH-responsive hydrogels have become effective and attractive materials for the controlled release of drugs at pre-determined destinations. In the present study, a novel hydrogel system based on Prunus armeniaca gum (PAG) and acrylic acid (AA) was prepared by a free radical mechanism using N, N-methylene bisacrylamide (MBA) as cross-linker and potassium persulfate (KPS) as initiator. A series of hydrogels varying PAG, AA, and MBA concentration was developed to determine the impact of these components. Formulated hydrogels were characterized for pH-responsive swelling, drug release, gel content, and porosity. Structural analysis was performed by FTIR, XRD, and SEM analysis. TGA study was applied to assess thermal stability. Oral acute toxicity and in vivo drug release were performed in rabbits. Hydrogels exhibited pH-dependent swelling and drug release. Swelling, drug loading and release, and porosity increased by increasing PAG and AA concentration while decreased by increasing MBA. The gel content of formulations was increased by increasing all three components. FTIR studies confirmed the development of copolymeric networks and the loading of drug. XRD studies revealed that hydrogels were amorphous, and the crystalline drug was changed into an amorphous form during loading. TGA results indicated that hydrogels were stable up to 600 °C. Acute oral toxicity results confirm that hydrogels were nontoxic up to a dose of 2 g/kg body weight in rabbits. The pharmacokinetic evaluation revealed that hydrogels prolonged the availability of the drug and the peak plasma concentration of the drug was obtained in 6 h as compared to the oral solution of the drug. Tramadol hydrochloride (THC) was used as a model drug. Hence, pH-responsive swelling and release, nontoxic nature and improved pharmacokinetics support that PAG-based hydrogels may be considered as potential controlled-release polymeric carriers.

Multipath Routing in Wireless Body Area Sensor Network for Healthcare Monitoring.

Mobility and low energy consumption are considered the main requirements for wireless body area sensor networks (WBASN) used in healthcare monitoring systems (HMS). In HMS, battery-powered sensor nodes with limited energy are used to obtain vital statistics about the body. Hence, energy-efficient schemes are desired to maintain long-term and steady connectivity of the sensor nodes. A sheer amount of energy is consumed in activities such as idle listening, excessive transmission and reception of control messages, packet collisions and retransmission of packets, and poor path selection, that may lead to more energy consumption. A combination of adaptive scheduling with an energy-efficient protocol can help select an appropriate path at a suitable time to minimize the control overhead, energy consumption, packet collision, and excessive idle listening. This paper proposes a region-based energy-efficient multipath routing (REMR) approach that divides the entire sensor network into clusters with preferably multiple candidates to represent each cluster. The cluster representatives (CRs) route packets through various clusters. For routing, the energy requirement of each route is considered, and the path with minimum energy requirements is selected. Similarly, end-to-end delay, higher throughput, and packet-delivery ratio are considered for packet routing.

Environmental remediation through various composite membranes moieties: Performances and thermomechanical properties.

Pollution harms ecosystems and poses a serious threat to human health around the world through direct or indirect effects on air, water, and land. The importance of remediating effluents is paramount to reducing environmental concerns. CO2 emissions are removed efficiently and efficaciously with mixed matrix membranes (MMMs), which are viable replacements for less efficient and costly membranes. In the field of membrane technology, MMMs are advancing rapidly due to their good separation properties. The selection of filler to be incorporated in mixed matrix membranes is very considered very important. There has been considerable interest in MOFs, carbon nanotubes (CNTs), ionic liquids (ILs), carbon molecular sieves (CMSs), sulfonated fillers (SFs), and layered silicates (LSs) as inorganic fillers for improving the properties of mixed matrix membranes. These fillers promise superb results and long durability for mixed matrix membranes based on them. The purpose of this review is to review different fillers used in MMMs for improving separation properties, limitations, and thermomechanical properties for environmental control and remediation.

Recent advancement challenges with synthesis of biocompatible hemodialysis membranes.

The focus of this study is to enhance the protein fouling resistance, hydrophilicity, biocompatibility, hemocompatibility and ability of the membranes and to reduce health complications like chronic pulmonary disease, peripheral vascular disease, cerebrovascular disease, and cardiovascular disease after dialysis, which are the great challenges in HD applications. In the current study, the PSF-based dialysis membranes are studied broadly. Significant consideration has also been provided to membrane characteristics (e.g., flowrate coefficient, solute clearance characteristic) and also on commercially available polysulfone HD membranes. PSF has gained a significant share in the development of HD membranes, and continuous improvements are being made in the process to make high flux PSF-based dialysis membranes with enhanced biocompatibility and improved protein resistance ability as the major issue in the development of membranes for HD application is biocompatibility. There has been a great increase in the demand for novel biocompatible membranes that offer the best performances during HD therapy, for example, low oxidative stress and low change ability of blood pressure.

Temporary epicardial pacing wires in isolated Coronary Artery Bypass Graft: Necessity or force of habit?

OBJECTIVE: To determine the frequency of Temporary epicardial pacing wires usage and its predictors in the immediate postoperative period in isolated coronary artery bypass graft surgery. METHODS: The longitudinal study was conducted at the Aga Khan University Hospital, Karachi, from September 2019 to August 2020, and comprised adult patients of either gender who underwent isolated coronary artery bypass graft in the Department of Cardiothoracic Surgery. Demographic, peri-operative and post-operative Temporary Epicardial Pacing Wires use data was extracted from patient's files and the institutional electronic database. Logistic regression models were built to explore predictors of Temporary epicardial pacing wires usage. Data was analysed using SPSS 22. RESULTS: Of the 322 cases evaluated, 27(8.4%) required the use of Temporary Epicardial Pacing Wires. Mean age of the patients requiring temporary epicardial pacing wires was 66.3±8.9 years compared to 58.7±8.9 years in those who did not require it (p<0.001), while the left ventricular ejection fraction percentage was 44.1±12.8 and 48.9±12.8 respectively (p=0.032). After adjusting for clinically plausible demographics and peri-operative variables, increasing age and low left ventricular ejection fraction were significantly associated with the use of temporary epicardial pacing wires in post-operative period of isolated coronary artery bypass graft patients (p<0.05). CONCLUSIONS: The frequency of temporary epicardial pacing wires usage in the post-operative period of coronary artery bypass graft was found to be low.

Ficus benghalensis as Potential Inhibitor of 5α-Reductase for Hair Growth Promotion: In Vitro, In Silico, and In Vivo Evaluation.

The screening of hair follicles, dermal papilla cells, and keratinocytes through in vitro, in vivo, and histology has previously been reported to combat alopecia. Ficus benghalensis has been used conventionally to cure skin and hair disorders, although its effect on 5α-reductase II is still unknown. Currently, we aim to analyze the phytotherapeutic impact of F. benghalensis leaf extracts (FBLEs) for promoting hair growth in rabbits along with in vitro inhibition of the steroid isozyme 5α-reductase II. The inhibition of 5α-reductase II by FBLEs was assessed by RP-HPLC, using the NADPH cofactor as the reaction initiator and Minoxin (5%) as a positive control. In silico studies were performed using AutoDock Vina to visualize the interaction between 5α-reductase II and the reported phytoconstituents present in FBLEs. Hair growth in female albino rabbits was investigated by applying an oral dose of the FBLE formulation and control drug to the skin once a day. The skin tissues were examined by histology to see hair follicles. Further, FAAS, FTIR, and antioxidants were performed to check the trace elements and secondary metabolites in the FBLEs. The results of RP-HPLC and the binding energies showed that FBLEs reduced the catalytic activity of 5α-reductase II and improved cell proliferation in rabbits. The statistical analysis (p < 0.05 or 0.01) and percentage inhibition (>70%) suggested that hydroalcoholic FBLE has more potential in increasing hair growth by elongating hair follicle's anagen phase. FAAS, FTIR, and antioxidant experiments revealed sufficient concentrations of Zn, Cu, K, and Fe, together with the presence of polyphenols and scavenging activity in FBLE. Overall, we found that FBLEs are potent in stimulating hair follicle maturation by reducing the 5α-reductase II action, so they may serve as a principal choice in de novo drug designing to treat hair loss.

Comparing sensitivity and specificity of pacemaker ID application and cardiac rhythm management device-finder application in identifying cardiac implantable electronic device manufacturer using chest radiograph - An observational study.

BACKGROUND: Smartphone-based applications to identify cardiac implantable electronic devices (CIED) are extremely useful in circumstances, where urgent device interrogation is needed, and a device identification card is not available. Few studies have provided insights regarding the utility of these applications. We have studied two widely available applications i.e., Pacemaker ID app (PMIDa) or Cardiac Rhythm Management Device-Finder (CRMD-f) to identify device manufacturers in CIEDs. METHODS: 547 patients who underwent CIED implantation from the year 2016-2020 in our institute were enrolled. There were 438 Medtronic and 109 St. Jude's devices. All chest radiographs were de-identified and resized into 225*225 pixels focusing on the CIED. PMIDa and CRMD-f applications were used to identify the CIED. Accuracy, sensitivity, specificity, negative predictive value, and positive predictive value for both applications were calculated and compared. RESULTS: Overall, CRMD-f application has higher specificity (93.58 vs. 82.5%) but lower sensitivity (53.6 vs. 55%) than PMIDa. The accuracy of both applications was comparable (61.6% vs. 60.5%). Accuracy varied with CIED model and type tested, and radiograph projection used. Accuracy is greatest with Cardiac-Resynchronization-Therapy (CRT) devices for both applications, followed by a single lead pacemaker. CONCLUSION: CRMD-f has higher accuracy and specificity for CIED manufacturer identification. Both PMIDa and CRMD-f are specific tools to identify CIED but have low sensitivity.

The determinants of environmental quality in the SAARC region: a spatial heterogeneous panel data approach.

In recent years, financial development, trade policies, and energy performance have attracted attention due to their behavior on environmental quality. Therefore, the current study examines the impact of financial development, trade openness, primary and renewable energy utilization, and economic growth on the ecological footprint in South Asian Association for Regional Cooperation (SAARC) countries from 1990 to 2017. This article progresses the proficiency of financial development by utilizing the comprehensive and multidimensional index of financial sector development based on their depth, access, and efficiency of their financial institutions and markets. In order to estimate the robust results, this study employed the cross-sectional dependency tests that allow the second-generation unit root, Westerlund cointegration, augmented mean group (AMG), error correction model (ECM), and Dumitrescu-Hurlin (D-H) panel non-causality tests. The results revealed a very weak effect of financial development in a panel of SAARC countries, while country-specific results reveal that financial development significantly enhances the pollution level in the case of Bangladesh and Sri Lanka. However, it improves the environmental quality in Nepal. Furthermore, trade openness only improves the environmental quality in the case of Nepal. Additionally, the findings explore that primary energy consumption enhances the ecological footprint in Bangladesh, Nepal, and Sri Lanka and reduces in case of Bhutan. On the contrary, renewable energy consumption significantly improves the environmental quality in all countries except Bangladesh. Finally, consistent with these findings, a number of suitable policy implications are expressed in the angle of SAARC economies.

Revealing empirical association among ecological footprints, renewable energy consumption, real income, and financial development: a global perspective.

This study quantifies the effect of real income, financial development, trade openness, and renewable energy consumption on the ecological footprint (EFP) of consumption for a panel data of 152 economies during the period 1990-2017. Several panel unit root tests validate that datasets are stationary. The findings from the Westerlund co-integration test depict that variables are co-integrated. The augmented mean group panel algorithm method is then applied to measure the long-run linkage between variables. The analysis outcomes show a negative and significant association between the EFP and real income per capita in the case of the higher-income group while remaining groups depict the other way round relationship. Further, openness and renewable energy consumption are also observed to reduce EFP in the groups of higher-income and upper-middle-income economies. Finally, financial development is observed to lessen environmental degradation in the case of the higher-income group. Similarly, the results of the Granger causality test based on the Dumitrescu-Hurlin panel provided evidence of varied causality relationship among the variables in different income groups. In addition, we also surpassed an impulse response and variance decomposition analysis that permitted to forecast the impact of concerned variables on environmental degradation during the selected period. Finally, the findings from the empirical analysis suggest that per capita economic growth will have an increasing effect on the EFP for the concerned income group except for higher-income countries in the future.

Long-term outcome of partial P450 side-chain cleavage enzyme deficiency in three brothers: the importance of early diagnosis.

OBJECTIVE: CYP11A1 mutations cause P450 side-chain cleavage (scc) deficiency, a rare form of congenital adrenal hyperplasia with a wide clinical spectrum. We detail the phenotype and evolution in a male sibship identified by HaloPlex targeted capture array. FAMILY STUDY: The youngest of three brothers from a non-consanguineous Scottish family presented with hyperpigmentation at 3.7 years. Investigation showed grossly impaired glucocorticoid function with ACTH elevation, moderately impaired mineralocorticoid function, and normal external genitalia. The older brothers were found to be pigmented also, with glucocorticoid impairment but normal electrolytes. Linkage studies in 2002 showed that all three brothers had inherited the same critical regions of the maternal X chromosome suggesting an X-linked disorder, but analysis of NR0B1 (DAX-1, adrenal hypoplasia) and ABCD1 (adrenoleukodystrophy) were negative. In 2016, next-generation sequencing revealed compound heterozygosity for the rs6161 variant in CYP11A1 (c.940G>A, p.Glu314Lys), together with a severely disruptive frameshift mutation (c.790_802del, K264Lfs*5). The brothers were stable on hydrocortisone and fludrocortisone replacement, testicular volumes (15-20 mL), and serum testosterone levels (24.7, 33.3, and 27.2 nmol/L) were normal, but FSH (41.2 µ/L) was elevated in the proband. The latter had undergone left orchidectomy for suspected malignancy at the age of 25 years and was attending a fertility clinic for oligospermia. Initial histology was reported as showing nodular Leydig cell hyperplasia. However, histological review using CD56 staining confirmed testicular adrenal rest cell tumour (TART). CONCLUSION: This kinship with partial P450scc deficiency demonstrates the importance of precise diagnosis in primary adrenal insufficiency to ensure appropriate counselling and management, particularly of TART.

Synthesis of 99m Tc-roxithromycin: A novel diagnostic agent to discriminate between septic and aseptic inflammation.

Roxithromycin is a second-generation macrolide antibiotic derived from erythromycin. In the current study, roxithromycin (ROX) was successfully labeled with technetium-99m for early diagnosis of bacterial infection and discrimination between septic and aseptic inflammation. The highest radiochemical purity of ≥95% was achieved by investigating different labeling parameters such as pH, ligand/reducing agent concentration, temperature, and amount of stabilizing agent. For this purpose, 0.3-0.5 mg ligand, 2-6 µg SnCl2 ·2H2 O as a reducing agent at basic pH (8-10 pH) and 2 mg mannitol used as a stabilizing agent, in the end, 370 MBq 99m Tc added into the reaction vials and incubated for a wide range of temperature (-4 to 65°C). The percent radiochemical purity of 99m Tc-roxithromycin was assessed with the help of the radio-thin-layer chromatography technique. The characterization studies were carried out using electrophoresis and Radio-HPLC techniques as well as saline stability and serum stability studies were also performed. Furthermore, biodistribution study was also performed in an inflamed animal model to discriminate between septic (heat-killed Staphylococcus aureus) and aseptic (turpentine oil) inflammatory lesions. The results were elaborated that 99m Tc-roxithromycin (99m Tc-ROX) was clearly bounded at the septic inflammation site (T/NT ratio of 7.08 ± 1.14) at 30 min postadministration, and maximum accumulation was seen in heart, lungs, liver, stomach, kidneys, and intestine. The results were suggested that 99m Tc-ROX might be used to discriminate between septic and aseptic inflammatory lesions at an early stage.

Synthesis and preclinical evaluation of bactericidal agent isolated from soil bacterium (Streptomyces): a novel diagnostic technique to enhance anticancer efficacy through radiosynthesis.

OBJECTIVES: Dactinomycin is a well-known antitumor-antibiotic drug isolated from soil bacterium Streptomyces, which exhibits broad-spectrum pharmacological and biochemical effects. In this study, dactinomycin was successfully labeled with technetium-99m for early diagnosis of bacterial infection and to discriminate it from acute inflammation. MATERIALS AND METHODS: Various labeling parameters such as pH, ligand concentration, reducing agent, and stabilizing agent were investigated. Radio-TLC technique was used to calculate percent radiochemical purity of radiopharmaceutical. Characterization studies were carried out using electrophoresis and radio-high-performance liquid chromatography techniques. Furthermore, saline and serum stability studies were performed to investigate biocompatibility. Biodistribution and scintigraphy studies were performed in infected and inflamed animal models to discriminate between bacterial infections (Escherichia coli and Staphylococcus aureus) and acute inflammations (heat-killed S. aureus). RESULTS: The results demonstrated that the highest radiochemical purity of at least 95% was achieved using 100-500 µg ligand and 3-8 µg SnCl2·2H2O as reducing agent at 4-9 pH. Technetium-99m-dactinomycin (Tc-DTN) was observed clearly bounded to the infection site having target/nontarget ratio 2.96±0.64 at 30 min after administration, which increased to 5.21±1.03 at 4 h after administration. Further accumulation was seen in heart, lungs, liver, stomach, kidneys, spleen, and intestine. An in-vitro cell-binding study was also performed, which showed high binding affinity of Tc-DTN with S. aureus-induced infectious lesions. CONCLUSION: Tc-DTN can easily be synthesized using standardized optimization conditions. The radiopharmaceutical has the highest accumulation potential at targeted site induced by S. aureus without any prominent in-vivo cytotoxicity. Tc-DTN may be used as a potential diagnostic agent to locate S. aureus-induced infection lesions at an early stage. Tc-DTN can successfully discriminate between infection and inflammatory models which cannot be achieved from other radiopharmaceuticals developed in the past few decades.

MCM - 2 and Ki - 67 as proliferation markers in renal cell carcinoma: A quantitative and semi - quantitative analysis

ABSTRACT Introduction/Background: Fuhrman nuclear grade is the most important histological parameter to predict prognosis in a patient of renal cell carcinoma (RCC). However, it suffers from inter-observer and intra-observer variation giving rise to need of a parameter that not only correlates with nuclear grade but is also objective and reproducible. Proliferation is the measure of aggressiveness of a tumour and it is strongly correlated with Fuhrman nuclear grade, clinical survival and recurrence in RCC. Ki-67 is conventionally used to assess proliferation. Mini-chromosome maintenance 2 (MCM-2) is a lesser known marker of proliferation and identifies a greater proliferation faction. This study was designed to assess the prognostic significance of MCM-2 by comparing it with Fuhrman nuclear grade and Ki-67. Material and Methods: n=50 cases of various ages, stages, histological subtypes and grades of RCC were selected for this study. Immunohistochemical staining using Ki-67(MIB-1, Mouse monoclonal antibody, Dako) and MCM-2 (Mouse monoclonal antibody, Thermo) was performed on the paraffin embedded blocks in the department of Morbid anatomy and Histopathology, University of Health Sciences, Lahore. Labeling indices (LI) were determined by two pathologists independently using quantitative and semi-quantitative analysis. Statistical analysis was carried out using SPSS 20.0. Kruskall-Wallis test was used to determine a correlation of proliferation markers with grade, and Pearson's correlate was used to determine correlation between the two proliferation markers. Results: Labeling index of MCM-2 (median=24.29%) was found to be much higher than Ki-67(median=13.05%). Both markers were significantly related with grade (p=0.00; Kruskall-Wallis test). LI of MCM-2 was found to correlate significantly with LI of Ki-67(r=0.0934;p=0.01 with Pearson's correlate). Results of semi-quantitative analysis correlated well with quantitative analysis. Conclusion: Both Ki-67 and MCM-2 are markers of proliferation which are closely linked to grade. Therefore, they can act as surrogate markers for grade in a manner that is more objective and reproducible.

MCM - 2 and Ki - 67 as proliferation markers in renal cell carcinoma: A quantitative and semi - quantitative analysis.

INTRODUCTION/BACKGROUND: Fuhrman nuclear grade is the most important histological parameter to predict prognosis in a patient of renal cell carcinoma (RCC). However, it suffers from inter-observer and intra-observer variation giving rise to need of a parameter that not only correlates with nuclear grade but is also objective and reproducible. Proliferation is the measure of aggressiveness of a tumour and it is strongly correlated with Fuhrman nuclear grade, clinical survival and recurrence in RCC. Ki-67 is conventionally used to assess proliferation. Mini-chromosome maintenance 2 (MCM-2) is a lesser known marker of proliferation and identifies a greater proliferation faction. This study was designed to assess the prognostic significance of MCM-2 by comparing it with Fuhrman nuclear grade and Ki-67. MATERIAL AND METHODS: n=50 cases of various ages, stages, histological subtypes and grades of RCC were selected for this study. Immunohistochemical staining using Ki-67(MIB-1, Mouse monoclonal antibody, Dako) and MCM-2 (Mouse monoclonal antibody, Thermo) was performed on the paraffin embedded blocks in the department of Morbid anatomy and Histopathology, University of Health Sciences, Lahore. Labeling indices (LI) were determined by two pathologists independently using quantitative and semi-quantitative analysis. Statistical analysis was carried out using SPSS 20.0. Kruskall-Wallis test was used to determine a correlation of proliferation markers with grade, and Pearson's correlate was used to determine correlation between the two proliferation markers. RESULTS: Labeling index of MCM-2 (median=24.29%) was found to be much higher than Ki-67(median=13.05%). Both markers were significantly related with grade (p=0.00; Kruskall-Wallis test). LI of MCM-2 was found to correlate significantly with LI of Ki-67(r=0.0934;p=0.01 with Pearson's correlate). Results of semi-quantitative analysis correlated well with quantitative analysis. CONCLUSION: Both Ki-67 and MCM-2 are markers of proliferation which are closely linked to grade. Therefore, they can act as surrogate markers for grade in a manner that is more objective and reproducible.