Monoclonal antibodies for copper-64 PET dosimetry and radioimmunotherapy

BACKGROUND: We previously described a two-antibody model of (64)Cu radioimmunotherapy to evaluate low-dose, solid-tumor response. This model was designed to test the hypothesis that cellular internalization is critical in causing tumor cell death by mechanisms in addition to radiation damage. The purpose of the present study was to estimate radiation dosimetry for both antibodies (mAbs) using positron emission tomography (PET) imaging and evaluate the effect of internalization on tumor growth. RESULTS: Dosimetry was similar between therapy groups. Median time to tumor progression to 1 g ranged from 7–12 days for control groups and was 32 days for both treatment groups (p < 0.0001). No statistically significant difference existed between any control group or between the treatment groups. MATERIAL AND METHODS: In female nude mice bearing LS174T colon carcinoma xenografts, tumor dosimetry was calculated using serial PET images of three mice in each group of either internalizing (64)Cu-labeled DOTA-cBR96 (DOTA = 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) or non-internalizing (64)Cu-labeled DOTA-cT84.66 from 3 to 48 h. For the therapy study, controls (n = 10) received saline, DOTA-cBR96 or DOTA-cT84.66. Treatment animals (n = 9) received 0.890 mCi of (64)Cu-labeled DOTA-cBR96 or 0.710 mCi of (64)Cu-labeled DOTA-cT84.66. Tumors were measured daily. CONCLUSIONS: PET imaging allows the use of (64)Cu for pre-therapy calculation of tumor dosimetry. In spite of highly similar tumor dosimetry, an internalizing antibody did not improve the outcome of (64)Cu radioimmunotherapy. Radio-resistance of this tumor cell line and copper efflux may have confounded the study. Further investigations of the therapeutic efficacy of (64)Cu-labeled mAbs will focus on interaction between (64)Cu and tumor suppressor genes and copper chaperones

HAEMOPHILIA B: CLINICAL MANIFESTATIONS AND COMPLICATIONS

ABSTRACT Background: Haemophilia B is X-linked recessive inherited disorder of factor IX deficiency. It is classified as severe, moderate and mild depending upon plasma levels of factor IX. The development of inhibitors is seen during treatment of haemophilia B against F-IX. This study was aimed to determine the frequency of different complications in haemophilia B patients. Patients and Methods: Total 45 patients of Haemophilia B already enrolled in the Haemophilia society of Pakistan Lahore chapter were included in this study. Clinical history and physical examinations were recorded on a pre designed proforma. Laboratory testing for establishment of diagnosis of haemophilia B and inhibitors of FIX was done. Results: Out of 45 patients, 10 (22.2%) had severe disease while 28 (62.2%) had moderate and 07 (15.6%) had mild disease. Twenty nine (64.4%) of patients with severe and moderate disease were diagnosed below 5 years of age while none with mild disease was diagnosed under 5 years of age. Arthropathy was the most frequently developing complication in patients 10 (100%) of severe Hemophilia B. Post circumcision bleeding was found to be the most common first episode of bleeding in patients of haemophilia B 29 (64.4%). Inhibitor against F-IX developed in only one patient of severe disease 1 (10%). Conclusion: Arthropathy is the commonest complication and circumcision is the first bleeding site in most of the haemophiliacs

Red Cell Alloimmunization in Repeatedly Transfused Cancer Patients

Background: To determine the frequency and specificities of red blood cells -antibodies in multi-transfused cancer patients. Methods: From total of 150 patients, 24 patients were diagnosed with haematological and 126 with non-haematological malignancies. Their medical records were reviewed and sorted by diagnosis, treatment medications, and past transfusion and/or treatment history. Only those patients were enrolled who had received at least five transfusions during the treatment. Antibody screening was performed by using commercial three-cell panel according to standardized methods. Allo-antibody was identified by using 11 cells panel. Results: Total patients with malignant disorders included 44 males and 106 females. Nine patients (6%) with non haematological malignancies possessed RBC allo-antibodies. The distribution of different allo-antibodies in positive patients was anti-D in 3 patients, anti-E in 2 patients while anti- Fya, anti Fyb, anti-Jka and anti-K were positive for 1 patient in each. The presence of allo-antibodies was statistically significant in patients with higher number of transfusions (p-value 0.001). Conclusion: Allo-antibodies against Rh system were most frequent. Incidence of RBCs allo-immunization increases with the number of transfusion and the number of donor exposures to the recipient

Comparison of changes in platelet count, mean platelet volume and swirling in stored platelet concentrates with and without platelet additive solution

Background: Stored platelet concentrates (PC) are in increasing demand for transfusion to patients with thrombocytopenia or disordered platelet function with active bleeding. Platelets are difficult to preserve in vitro for longer period of time. Different platelet additive solutions (PAS) have been formulated which can increase the survival period of PC during storage. Study Design: Comparative study. Materials and Methods: The study was carried out to compare in vitro changes in platelet indices [platelet count and mean platelet volume (MPV)] and extent of swirling in stored PC with and without PAS stored for 10 day. Results: This study included 42 samples of random donor platelet concentrates, divided into two groups. In one group, PC were stored without PAS while in other group PAS was added to PC before storage. Changes in platelets count and MPV were monitored by using automated hematology cell analyzer, in both groups on day 0, 3, 5, 7, 8, 9, and 10. Bacterial cultures were also applied to detect bacterial contamination of PC. The results showed that mean platelet count of PC stored without PAS was 5.6 + 0.1 x 1010/L on day 0 and it was 5.5 + 0.5 x 1010/L in PC stored with PAS. The difference between the two groups started becoming statistically significant by day 7. Platelet count was significantly lower in PC without PAS as compared to the PC stored with PAS (P value ≤ 0.001). The difference in the mean platelet volume (MPV) between two groups was highly significant during 10 days storage (P value ≤ 0.001). Swirling was better seen in PC stored in PAS as compared to PC stored without PAS during 10 days storage. Blood cultures were applied on 42 samples on days 0, 5 and 10. Bacterial contamination occurred in few samples in both groups, those samples were not processed further. Conclusion: This study showed that the use of PAS in platelet concentrate storage bags increased the shelf life and viability of platelets as compared to those without PAS

Serum hepcidin levels in patients with end-stage renal disease on hemodialysis

Patients on hemodialysis (HD) are usually anemic because of defective erythropoeisis. Hepcidin is a polypeptide that regulates iron homeostasis and could serve as an indicator of functional iron deficiency in patients with end-stage renal disease (ESRD); this may also aid in the assessment of patient′s response to erythropoietin (EPO). The present study was directed to investigate serum levels of hepcidin, iron status and inflammation markers such as C-reactive protein (CRP) in patients with ESRD on maintenance HD and to observe the correlation of serum hepcidin with conventional iron and inflammatory markers. A total of 42 patients of both sexes on maintenance HD and EPO therapy were enrolled; 42 ageand sex-matched healthy subjects were included as controls. Laboratory tests including complete blood count, serum hepcidin, total iron binding capacity (TIBC), serum ferritin, serum iron and CRP were performed. Serum hepcidin levels were significantly higher in patients with ESRD than in the control group (18.2 ± 2.8 ng/mL and 8.5 ± 2.3 ng/mL, respectively P = 0.000). The hemoglobin, hematocrit, serum iron, TIBC and transferrin saturation levels in the patient group were significantly lower than in the control group. Higher hepcidin levels were found in EPO non-responders (19.6 ± 2.4 ng/mL) while lower levels (16.9 ± 2.5 ng/mL) were seen in responders (P = 0.001). A positive and significant correlation was observed between the values of serum hepcidin and CRP. Our study indicates that higher hepcidin levels are found in ESRD patients on HD and in those not responding to EPO. Our findings suggest that hepcidin might play a role in the pathophysiology of anemia associated with chronic diseases as well as EPO resistance

Enhancing oral health diagnosis and treatment with artificial intelligence in dentistry

This study aims to explore the potential of artificial intelligence (AI) in enhancing oral health diagnosis and treatment in the field of dentistry. With a sample size of 100 individuals from the population of Lahore, this research investigates the efficacy of AI technologies in revolutionizing oral healthcare practices. The traditional approach to oral health diagnosis and treatment heavily relies on subjective human judgment, leading to variations in diagnoses and treatment plans. By leveraging AI algorithms, this study demonstrates how machine learning and deep learning techniques can assist dentists in making accurate and efficient diagnoses, resulting in improved treatment outcomes. The research involves the development of an AI model trained on a dataset comprising dental records and associated clinical information. Through the analysis of this dataset, the AI model learns to recognize patterns, identify abnormalities, and predict potential oral health issues. By integrating this model into dental practices, dentists can make informed decisions based on data-driven insights, ultimately leading to enhanced patient care. Furthermore, this study evaluates the potential challenges and ethical considerations associated with implementing AI in dentistry.&nbsp

Determination of IL-6 Gene Promoter Polymorphism in Patients with Hepatitis C and Its Impact on RNA Secondary Structure

Background and Objectives: A polymorphism in the promoter region of the IL-6 gene would influence the level of IL-6 expression in patients with HCV, resulting in a pro-inflammatory response. Few studies have shown the association between −174G>C (rs1800795) and −1363G>T (rs2069827) polymorphisms and HCV infection, and their results have been contradictory. There are no data published in our population to study such an IL-6 stimulus against HCV infection and its impact on RNA secondary structure. Therefore, we isolated human subjects from the province of Punjab, Pakistan. The objective was to screen for IL-6 gene promoter polymorphisms −174G/C and −1363G/T and those correlated with serum concentrations of IL-6 in patients with HCV and compared with a control. Materials and Methods: In conventional PCR, measurement of serum IL-6 by CLIA and statistical analysis were performed to observe the genotype association studies. By integrating bioinformatics and computational tools, our study aimed to provide a comprehensive understanding of how variations in the promoter region of IL-6 may have functional implications on gene expression. Results: The −174G>C and −1363G>T genotypes in the promoter region of patients with HCV were in strong allelic association (Δ = 0.97, p Conclusions: Based on the data, it can be inferred that IL-6 gene promoter polymorphisms are important in the dysregulation of IL-6 levels in patients with HCV

Metabolic Profiling by GC-MS, In Vitro Biological Potential, and In Silico Molecular Docking Studies of Verbena officinalis

Verbena officinalis L. is a traditionally important medicinal herb that has a rich source of bioactive phytoconstituents with biological benefits. The objective of this study was to assess the metabolic profile and in vitro biological potential of V. officinalis. The bioactive phytoconstituents were evaluated by preliminary phytochemical studies, estimation of polyphenolic contents, and gas chromatography-mass spectrometry (GC-MS) analysis of all fractions (crude methanolic, n-hexane, ethyl acetate, and n-butanol) of V. officinalis. The biological investigation was performed by different assays including antioxidant assays (DPPH, ABTS, CUPRAC, and FRAP), enzyme inhibition assays (urease and &alpha;-glucosidase), and hemolytic activity. The ethyl acetate extract had the maximum concentration of total phenolic and total flavonoid contents (394.30 &plusmn; 1.09 mg GAE&middot;g&minus;1 DE and 137.35 &plusmn; 0.94 mg QE&middot;g&minus;1 DE, respectively). Significant antioxidant potential was observed in all fractions by all four antioxidant methods. Maximum urease inhibitory activity in terms of IC50 value was shown by ethyl acetate fraction (10 &plusmn; 1.60 &micro;g mL&minus;1) in comparison to standard hydroxy urea (9.8 &plusmn; 1.20 &micro;g&middot;mL&minus;1). The n-hexane extract showed good &alpha;-glucosidase inhibitory efficacy (420 &plusmn; 20 &micro;g&middot;mL&minus;1) as compared to other extract/fractions. Minimum hemolytic activity was found in crude methanolic fraction (6.5 &plusmn; 0.94%) in comparison to positive standard Triton X-100 (93.5 &plusmn; 0.48%). The GC-MS analysis of all extract/fractions of V. officinalis including crude methanolic, n-hexane, ethyl acetate, and n-butanol fractions, resulted in the identification of 24, 56, 25, and 9 bioactive compounds, respectively, with 80% quality index. Furthermore, the bioactive compounds identified by GC-MS were analyzed using in silico molecular docking studies to determine the binding affinity between ligands and enzymes (urease and &alpha;-glucosidase). In conclusion, V. officinalis possesses multiple therapeutical potentials, and further research is needed to explore its use in the treatment of chronic diseases