Effects of graded levels of dietary microbial 6-phytase on performance, intestinal histomorphology, caecal microbial population and short-chain fatty acid composition of Lohmann white-classics.

1. This study was conducted to determine the effects of graded levels of phytase on the performance, egg quality and gut health of white laying hens.2. Treatments consisted of a negative control (NC) diet containing 0.14% available phosphorus (avP), positive control (PC) diet containing 0.35% avP provided via dicalcium phosphate (DCP) and DCP replaced in the PC by with three graded levels of phytase derived from Komagataella phaffii at 500 (PC-500), 750 (PC-750) and 1000 (PC-1000) FTU/kg which provided 0.176%, 0.188% and 0.200% of avP, respectively.3. Egg production, feed intake, feed conversion ratio and jejunal morphometry were negatively affected in NC-fed birds (p < 0.05). Considering the whole period, birds fed a diet supplemented with graded levels of phytase shared the same egg production and feed intake levels with PC birds (p < 0.05). Feed conversion ratio was significantly lowered by 4.9%, 1.6% and 7.6% in hens fed on diets PC-500, PC-750 and PC-1000, respectively compared to those fed the PC (p < 0.05).4. Neither of the dietary treatments affected cracked eggs, dirty eggs, eggshell breaking strength and eggshell thickness. Dietary supplementation of phytase significantly increased villus surface area by 15%, 36% and 40% in PC-500, PC-750 and PC-1000 birds, respectively compared to PC (p < 0.05).5. A significant increase in lactobacillus count was observed in line with increasing the level of phytase (p < 0.05). Dietary treatments had no effect on the caecal coliform or aerobic populations. Furthermore, phytase supplementation significantly increased the concentrations of total caecal short-chain fatty acid (SCFA; p < 0.01).6. In conclusion, along with improving performance parameters, the inclusion of phytase in laying hen diets can ameliorate intestinal morphology and stimulate caecal microflora and increase SCFA concentrations.

A review of vaginal hysterectomies in the gynaecology department of a tertiary care hospital.

This retrospective cohort study analysed the trends and complications of vaginal hysterectomy conducted at Services Hospital, Lahore, from January 1, 2015 to December 31, 2020. Demographics, indications, surgery duration, complications (haemorrhage, urological or rectal problems, infection), and hospital stay were recorded. Out of 819 hysterectomies performed for benign gynaecological conditions, 112 (13.68%) were vaginal hysterectomies. Non-descent vaginal hysterectomy (NDVH) accounted for 33(29.46%) and uterine prolapse for 79(70.53%) of the cases. Mean age was 52.35±8.74 years, parity was 5.01±1.32, intraoperative haemorrhage was 796.87±450.1 ml, surgery duration was 48.61±12.28 minutes, and hospital stay was 2.58±0.41 days. Complications occurred in 19(16.97%) of the cases, while 93(83.03%) cases had no complications. Outcomes were comparable between NDVH and vaginal hysterectomy for prolapse (p=0.552). This indicates that vaginal hysterectomy is a safe procedure with minimal complications and quick recovery for uterine prolapse and non-descent uterus. However, a declining trend was observed over the study period.

Advancing Research on Proprotein Convertase Subtilisin/Kexin Type 9 Inhibitors: A Scientometric Analysis.

Atherosclerosis is characterised by the accumulation of fatty deposits and plaque as a result of a continuously high level of low-density lipoprotein cholesterol (LDL-C) in the blood. The primary objective of this research is to assess the current status of knowledge, research endeavours and developmental trajectories about proprotein convertase subtilisin/kexin type 9 (PCSK9) inhibitors in correlation with atherosclerosis treatment. Additionally, this study aims to compile bibliometric and scientometric investigations within this domain through rigorous scientometric analysis. Analysing the bibliometric landscape and global research trends associated with PCSK9 inhibitors can contribute valuable insights into comprehending atherosclerosis. This is exemplified by examining publications within the Web of Science Core Collection (WOSCC) database from 2008 to 2022. Citespace was used for frequency, co-occurrence, co-citation, grouping and burst analysis, and Microsoft Excel was used to manage descriptive datasets. Eight hundred eighty-five publications available from WOSCC database between the years 2008 and 2022 were extracted and examined. Over the period, 3,138 collaborating institutions from 87 countries, a staggering 7,750 writers involved and 325 distinct journals published about PCSK9 inhibitors studies. Among authors, Sabatine et al. and the journal The New England Journal of Medicine has had the most significant impact. Lipid-lowering therapy and bempedoic acid are the most prominent topical clusters associated with PCSK9 inhibitors, and the most often used keywords are efficacy, safety and PCSK9 inhibitors. We believe this is the first comprehensive analysis of PCSK9 inhibitors research and publications conducted using Scientometric. These results demonstrate the nascence of PCSK9 inhibitors research. They may encourage a wide range of stakeholders, particularly early career researchers from various disciplines, to work together in the future.

Effect of Cypermethrin on Blood Hematology and Biochemical Parameters in Fresh Water Fish Ctenopharyngodon idella (Grass Carp).

The insecticide, cypermethrin, adversely affects biochemical parameters in blood and behavior in grass carp (Ctenopharyngodon idella). Fish were obtained from hatchery, reared in the laboratory. Different concentration of cypermethrin were applied. Blood was collected and hematological and biochemical parameters were measured. Biochemical parameters such as, protein levels, cholesterol, phosphorous and calcium in both acute and chronically cypermethrin treated groups decreased, with increasing exposure time from 24h to 15 days with more pronounced effects in the acute groups. Increased glucose, urea, serum glutamic pyruvic transaminase (SGPT), creatinine, and lactate dehydrogenase (LDH) levels were found in both acute and chronic groups with the increasing exposure time. Hematological parameters, such as red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MHCH), and red cell distribution width (RDW) were significantly reduced in both groups as the exposure time increases. However, the numbers of white blood cells (WBC) and platelets were increased. This study established both the acute and chronic toxicity of cypermethrin in grass carp, which likely occurs secondary to altered biochemical and blood parameters.

Evaluation of Anti-clotting and thrombolytic potential of the aqueous-methanolic extract of Jasminum sambac.

This study was done to investigate the anti-clotting and thrombolytic potential of ariel part of Jasminum sambac (L.). Healthy male rabbits were divided into five (5) groups with each group containing six (6) animals. Aqueous-methanolic extract of the plant was prepared and given at different doses of 200, 300 and 600mg/kg to three groups in comparison to negative and positive control groups. The aqueous-methanolic extract showed a dose-dependent increase in the activated partial thromboplastin (APTT) and prothrombin time (PT), bleeding time (BT) and clotting time (CT) (p<0.05). Warfarin 2mg/kg was used as the standard. The plant extract also showed significant (p<0.05) clot lysis in comparison with standard urokinase. Moreover, it also prolonged the ADP induced platelet adhesion at doses of 200, 300 and 600µg/mL dependently. HPLC analysis showed rutin, quercetin, salicylic acid and ascorbic acid as vital phytoconstituents in aqueous-methanolic extract. Anticoagulant and thrombolytic effect of Jasminum sambac justified its therapeutic utility in cardiovascular disorders and this may be due to the presence of salicylic acid, rutin and quercetin in the extract.

Maternal near-miss, mortality and their correlates at a tertiary care hospital.

OBJECTIVE: To determine the frequency and causes of maternal near-miss and mortality among pregnant women. METHODS: The cross-sectional prospective study was conducted from January 2016 to December 2018 at the Services Hospital, Lahore, Pakistan, and comprised all near-miss cases admitted in the gynaecology department. The World Health Organisation criterion was used to identify the near-miss cases. Primary outcome measures were frequency and causes of near-miss and the ratio between maternal mortality and near-miss cases. Secondary outcome measures were delays, need for massive blood transfusion, intensive care unit admission, obstetric hysterectomy and hospital stay >7 days. Data was analysed using SPSS 23. RESULTS: Of the 10,739 live births, there were 344(3.2%) complications resulting in 305(2.8%) near-miss cases and 29(0.3%) maternal deaths. Frequency of near miss was 28.4 per 1000 live births and the ratio between maternal mortality and near-miss cases was 1:10.5. There were 215(62.5%) unbooked patients and 23(6.7%) of them died (p<0.001). Among the near-miss cases, haemorrhage accounted for 150(49.18%), hypertensive disorders 102(33.44%), cardiac disease 25(8.28%) and infection 12(3.97%). Maternal mortality was significantly low for haemorrhage, hypertension, sepsis and cardiac disease (p<0.001). Massive blood transfusion was given to 64(20.98%) patients, 48(15.74%) underwent hysterectomy, and 98(32.13%) required admission to intensive care unit. First and second delays were seen in 240(78.6%) near-miss and 25(86.2%) mortality cases (p<0.001). CONCLUSIONS: Haemorrhage and hypertension were found to be major reasons for near-miss cases, but timely intervention could prevent mortality.

Gynecological malignancies at tertiary care hospital, Pakistan: A five-year review.

BACKGROUND & OBJECTIVE: Gynecological malignancies are important cause of female morbidity and mortality. They pose significant burden on health resources in low middle-income countries. Data on presentation and risk factors can help in early identification and reduce this burden. Our objective was to evaluate frequency, stage of presentation and risk factors of gynecological malignancies in a tertiary care setting. METHODS: It was cross sectional study done in Gynecology Department, Services Institute of Medical Sciences, Services Hospital, Lahore from January 2015- December 2019. The records of the patients were retrospectively reviewed to include all cases of gynecologic malignancies. Demographic information, frequency, risk factors, symptoms, grade and stage of tumor was collected. RESULTS: There were 122 patients diagnosed with gynecological malignancy during the study period. Ovarian cancer was seen in 60 (49.18%) patients followed by cervical cancer in 29(23.7%), endometrial cancer 27(22.1%) and vulva 06(4.9%). Mean age for all cancers was 51±12.7 to 55±9.3 except cervical cancer which was seen in 43±8.9 years. Patients with ovarian cancer had significantly more hypertension and diabetes (p<0.05). Heavy menstrual bleeding and postmenopausal bleeding was significantly seen in patients of endometrial and cervical cancer (p<0.05). Abdominal symptoms of pain, mass and distension were seen in patients with ovarian cancer (p<0.05). Majority patients presented in advanced stage. Among ovarian cancer, 52/60(86.6%) were epithelial in origin while 25(86.2%) cervical cancer and all vulva cancers were squamous cell carcinoma. CONCLUSION: Ovarian cancer was commonest gynecological malignancy followed by cervical cancer. Late presentation with advanced stage was seen in majority of all cancers.

Phenotypic and molecular characterization of virulence factors of extra-intestinal pathogenic Escherichia coli isolated from patients of Peshawar, Pakistan.

Extra-Intestinal Escherichia coli (ExPEC) are important cause of Urinary Tract Infections (UTIs) and systemic infections. The purpose of this study was to investigate numerous ExPEC bacterial isolates for phenotypic virulence characteristics including hemolytic activity and resistance pattern and to observe their association with genetic traits via Polymerase Chain Reaction (PCR). A total of 367 ExPEC isolates were collected from patients admitted in Khyber Teaching Hospital (KTH) Peshawar, Pakistan. Standard techniques were used for identification of isolates, determination of hemolytic potential and antimicrobial susceptibility testing. PCR was used for screening of virulence genes using specific primers. A total of 367 ExPEC isolates were characterized, among which 62.7, 24.3, 7.1 and 6% were isolated from urine, pus, sputum and wound specimens, respectively. Majority of the isolates (82.8%) were hemolysin positive. Multi drug resistance pattern was shown by 41% of the isolates and harbored at least one virulence gene (71.7%), of which sat was the most prevalent (64.3%). The highest resistance was found to cefotaxime (99.2%), ampicillin (97.5%) and aztreonem (89.6%). 15 different virulence genes combinations were observed in the current study. A total of 16 virotypes (15 of positive virulence genes and one of no virulence gene) were observed in the current study. The current investigation showed a high prevalence of sat and hlyA genes among ExPEC isolate, suggesting a role of these genes in the pathogenesis of ExPEC.

Designing of potent inhibitors for metallo-beta-lactamases producing Escherichia coli in molecular specification hereto.

The rapid spread of Metallo-ß-Lactamases producing Gram-negative bacteria in Pakistan is alarming and novel inhibitors with multi inhibition potential are required. In the current study, an effort was made to identify the resistance genes of MBLs producing E. coli and single inhibitor was designed having the potential to block all resistant proteins. Results showed that out of 573 clinical isolates, 14.1% MBLs producers have NDM-1 (27.2%) and VIM (13.6%) gene. The isolates were resistant to MEM, AMP, AMC, FEP, CTX, LEV and ATM, while effective antibiotics were TGC, CO, FOS and AK with MICs ranging from 4 to >32µg/ml. RECAP synthesis was used for de-novo discovery of 1000 inhibitors and protein crystal structures were retrieved from PDB. Active sites were identified in each protein and to improve ADMET properties, Lipinski's rules of five was applied. Placement of the ligand was done by London dG algorithm implemented in MOE. For final refinement, GBVI/WSA dG algorithm was used. Based on docking score, visual inspection of ligands interaction with key residues, binding affinity and binding energy of ligands with proteins, 10 compounds were selected for MBLs proteins which presented best ADMET properties, binding energy and affinity than the reported ones.

Alterations of elastin in female reproductive tissues arising from advancing parity.

Female reproductive tissues undergo significant alterations during pregnancy, which may compromise the structural integrity of extracellular matrix proteins. Here, we report on modifications of elastic fibers, which are primarily composed of elastin and believed to provide a scaffold to the reproductive tissues, due to parity and parturition. Elastic fibers from the upper vaginal wall of virgin Sprague Dawley rats were investigated and compared to rats having undergone one, three, or more than five pregnancies. Optical microscopy was used to study fiber level changes. Mass spectrometry, 13C and 2H NMR, was applied to study alterations of elastin from the uterine horns. Spectrophotometry was used to measure matrix metalloproteinases-2,9 and tissue inhibitor of metalloproteinase-1 concentration changes in the uterine horns. Elastic fibers were found to exhibit increase in tortuosity and fragmentation with increased pregnancies. Surprisingly, secondary structure, dynamics, and crosslinking of elastin from multiparous cohorts appear similar to healthy mammalian tissues, despite fragmentation observed at the fiber level. In contrast, elastic fibers from virgin and single pregnancy cohorts are less fragmented and comprised of elastin exhibiting structure and dynamics distinguishable from multiparous groups, with reduced crosslinking. These alterations were correlated to matrix metalloproteinases-2,9 and tissue inhibitor of metalloproteinase-1 concentrations. This work indicates that fiber level alterations resulting from pregnancy and/or parturition, such as fragmentation, rather than secondary structure (e.g. elastin crosslinking density), appear to govern scaffolding characteristics in the female reproductive tissues.

Antibacterial evaluation of silver nanoparticles synthesized from lychee peel: individual versus antibiotic conjugated effects.

This paper describes the extracellular synthesis of silver nanoparticles from waste part of lychee fruit (peel) and their conjugation with selected antibiotics (amoxicillin, cefixim, and streptomycin). FTIR studies revealed the reduction of metallic silver and stabilization of silver nanoparticles and their conjugates due to the presence of CO (carboxyl), OH (hydroxyl) and CH (alkanes) groups. The size of conjugated nanoparticles varied ranging from 3 to 10 nm as shown by XRD. TEM image revealed the spherical shape of biosynthesized silver nanoparticles. Conjugates of amoxicillin and cefixim showed highest antibacterial activity (147.43 and 107.95%, respectively) against Gram-negative bacteria i.e. Alcaligenes faecalis in comparison with their control counterparts. The highest reduction in MIC was noted against Gram-positive strains i.e. Enterococcus faecium (75%) and Microbacterium oxydans (75%) for amoxicillin conjugates. Anova two factor followed by two-tailed t test showed non-significant results both in case of cell leakage and protein estimation between nanoparticles and conjugates of amoxicillin, cefixime and streptomycin. In case of MDA release, non-significant difference among the test samples against the selected strains. Our study found green-synthesized silver nanoparticles as effective antibacterial bullet against both Gram positive and Gram negative bacteria, but they showed a more promising effect on conjugation with selected antibiotics against Gram negative type.

The ADRIFT study - Assessing Diabetes Distress and its associated factors in the Pakistani population.

OBJECTIVE: To assess diabetes distress and its associated factors in Pakistani population.. METHODS: The cross-sectional study was conducted at Pakistan Institute of Medical Sciences, Islamabad, Pakistan, from July to December 2017, and comprised patients of diabetes type 2. After noting down demographic and clinical parameters, diabetes distress of the subjects was measured by applying the 17-item diabetes distress scale which also assesses sub domains like emotional burden, physicianrelated distress, regimen-related distress and interpersonal distress. SPSS 20 was used to analyse data. RESULTS: There were 349 subjects with a mean age of 53.14±11.77 years, mean diabetes duration of 8.36±6.64 years and a mean glycated haemoglobin value of 9.05±1.93%. Mean overall diabetes distress score was 2.55±0.75, signifying moderate distress. Overall, prevalence of diabetes distress was found among 266(76.2%) subjects;164(47%) moderate and 102(29.2%) high level distress. Emotional burden was most substantially elevated, with 296(84.8%) patients reporting moderate to high levels. Total diabetes distress was significantly related to demographic background (p<0.0001), education level (p=0.015), monthly income, frequency of administration of medication, adherence to medical treatment (p<0.05), number of complications (p<0.05) and overall glycaemic control (p<0.001). CONCLUSIONS: Modifiable factors, such as frequency of medication and compliance to treatment, should be addressed with the aim of decreasing diabetes distress and improve glycaemic control..

Standardized programming to reduce the burden of inappropriate therapies in implantable cardioverter defibrillators - Single centre follow up results.

BACKGROUND: Current algorithms and device morphology templates have been proposed in current Implantable Cardioverter-Defibrillators (ICDs) to minimize inappropriate therapies (ITS), but this has not been completely successful. AIM: Assess the impact of a deliberate strategy of using an atrial lead implant with standardized parameters; based on all current ICD discriminators and technologies, on the burden of ITS. METHOD: A retrospective single-centre analysis of 250 patients with either dual chamber (DR) ICDs or biventricular ICDs (CRTDs) over a (41.9 ± 27.3) month period was performed. The incidence of ITS on all ICD and CRTD patients was chronicled after the implementation of standardized programming. RESULTS: 39 events of anti-tachycardial pacing (ATP) and/or shocks were identified in 20 patients (8% incidence rate among patients). The total number of individual therapies was 120, of which 34% were inappropriate ATP, and 36% were inappropriate shocks. 11 patients of the 250 patients received ITS (4.4%). Of the 20 patients, four had ICDs for primary prevention and 16 for a secondary prevention. All the episodes in the primary indication group were inappropriate, while seven patients (43%) of the secondary indication group experienced inappropriate therapies. CONCLUSIONS: The burden of ITS in the population of patients receiving ICDs was 4.4% in the presence of atrial leads. The proposed rationalized programming criteria seems an effective strategy to minimize the burden of inappropriate therapies and will require further validation.

Impact of maternal respiratory infections on low birth weight - a community based longitudinal study in an urban setting in Pakistan.

BACKGROUND: The health of mothers and their newborns is intricately related. The weight of the infant at birth is a powerful predictor of infant growth and survival, and is considered to be partly dependent on maternal health and nutrition during pregnancy. We conducted a longitudinal study in an urban community within Karachi to determine maternal predictors of newborn birth weight. METHODS: Four hundred pregnant women were enrolled in the study during the period 2011-2013. Data related to symptoms of acute respiratory illness (fever, cough, difficulty breathing, runny nose, sore throat, headache, chills, and myalgia/lethargy) in the pregnant women were collected weekly until delivery. Birth weight of the newborn was recorded within 14 days of delivery and the weight of <2.5 kg was classified as low birth weight (LBW). RESULTS: A total of 9,853 symptom episodes were recorded of fever, cough, difficulty breathing, runny nose, sore throat, headache, chills, myalgias/lethargy in the enrolled pregnant women during the study. Out of 243 pregnant women whose newborns were weighed within 14 days of birth, LBW proportion was 21% (n = 53). On multivariate analysis, independent significant risk factors noted for delivering LBW babies were early pregnancy weight of < 57.5 kg [odds ratio adjusted (ORadj) = 5.1, 95% CI: (1.3, 19.9)] and gestational age [ORadj = 0.3, 95% CI (0.2, 0.7) for every one week increase in gestational age]. Among mothers with high socioeconomic status (SES), every 50-unit increase in the number of episodes of respiratory illness/100 weeks of pregnancy had a trend of association with an increased risk of delivering LBW infants [ORadj = 1.7, 95% CI: (1.0, 3.1)]. However, among mothers belonging to low SES, there was no association of the number of episodes of maternal respiratory illness during pregnancy with infants having LBW [ORadj = 0.9, 95% CI: (0.5, 3.5)]. CONCLUSIONS: While overall respiratory illnesses during pregnancy did not impact newborn weight in our study, we found this trend in the sub-group of mothers belonging to the higher SES. Whether this is because in mothers belonging to lower SES, the effects of respiratory illnesses were overshadowed by other risk factors associated with poverty need to be further studied.

Self-evaluation of ethical review committees functioning at Foundation University Medical College (FUMC) through structured constitution-practice-outcome (CPO) assessment model.

OBJECTIVE: To assess the operational efficiency of the ethical review committee of a medical college. METHODS: This study was conducted at the Foundation University Medical College, Islamabad, Pakistan, from 2012 to 2014. On the basis of literature review, methods for assessment of various features of ethical review committee were studied. A constitution-practice-outcome measurement model for evaluation of ethical review committee assessment process was developed. Data submitted to ethical review committee since its constitution was extracted and quantitatively analysed. RESULTS: The ethical review committee comprised 14 members, including 4(28.6%) permanent and 10(71.4%) rotating clinical, basic sciences and non-medical members. As many as 45 research protocols were submitted, with submission frequency of 8(17.8%), 12(26.7%) and 25(55.5%) per year respectively, and issued ethical approval certificates within a mean duration of 7.2±3.2 days from the time of first submission to final notification. Issues looked into were according to World Health Organisation guidelines. Standard review was done on 29(64.4%) studies and expedited on 16(35.5%). In addition, 24(53.3%) protocols needed resubmission. Only 2(4.4%) protocols were not approved. The number of issues raised for resubmission was 71. Main reasons for resubmission were found to be incomplete documents 26(36.6%), invalid informed consent forms 12(16.9%) and negligence in maintaining confidentiality of study participants 9(12.7%). CONCLUSIONS: Ethical review committee with its limited resources was fulfilling its founding objectives as depicted by constitution-practice-outcome model.

Pregnancy-Induced Dynamical and Structural Changes of Reproductive Tract Collagen.

The tissues and organs of the female reproductive tract and pelvic floor undergo significant remodeling and alterations to allow for fetal growth and birth. In this work, we report on a study of the alterations of murine reproductive tract collagen resulting from pregnancy and parturition by spectrophotometry, histology, and (13)C, (2)H nuclear magnetic resonance (NMR). Four different cohorts of rats were investigated that included virgin, multiparous, two- and fourteen-day postpartum primiparous rats. (13)C CPMAS NMR revealed small chemical shift differences across the cohorts. The measured H-C internuclear correlation times indicated differences in dynamics of some motifs. However, the dynamics of the major amino acids, e.g., Gly, remained unaltered with respect to parity. (2)H NMR relaxation measurements revealed an additional water reservoir in the postpartum and multiparous cohorts pointing to redistribution of water due to pregnancy and/or parturition. Spectrophotometric measurements indicated that the collagen content in virgin rats was highest. Histological analysis of the upper vaginal wall indicated a signature of collagen fiber dissociation with smooth muscle and a change in the density of collagen fibers in multiparous rats.

Dynamic patient grouping and prioritization: a new approach to emergency department flow improvement.

The demand on emergency departments (ED) is variable and ever increasing, often leaving them overcrowded. Many hospitals are utilizing triage algorithms to rapidly sort and classify patients based on the severity of their injury or illness, however, most current triage methods are prone to over- or under-triage. In this paper, the group technology (GT) concept is applied to the triage process to develop a dynamic grouping and prioritization (DGP) algorithm. This algorithm identifies most appropriate patient groups and prioritizes them according to patient- and system-related information. Discrete event simulation (DES) has been implemented to investigate the impact of the DGP algorithm on the performance measures of the ED system. The impact was studied in comparison with the currently used triage algorithm, i.e., emergency severity index (ESI). The DGP algorithm outperforms the ESI algorithm by shortening patients' average length of stay (LOS), average time to bed (TTB), time in emergency room, and lowering the percentage of tardy patients and their associated risk in the system.

Pakistani children's experiences of growing up with beta-thalassemia major.

In this study, we explored the lived experiences of children with beta-thalassemia major (ß-TM). We considered children as experts on their experiences in contrast to the prevalent approach of asking parents or other adults about children's perspectives. The sample consisted of 12 children aged 8 to12 years. There were two stages to data collection. In Stage 1 we employed two focus group discussions and two role plays and analyzed the data thematically. This directly informed Stage 2, consisting of 12 in-depth interviews subjected to interpretative phenomenological analysis. From our findings we show that living with ß-TM involves a continuous struggle between feelings of being different and strategies to minimize these differences to strive for normalcy. We suggest that understanding the experiences of living with ß-TM from children's perspectives can provide unique insights into their experiences, which can fill the gap in the existing, predominantly adult-oriented research on chronic illness.

Hispidulin: a promising anticancer agent and mechanistic breakthrough for targeted cancer therapy.

Cancer is a complex disease characterized by dysregulated cell growth and division, posing significant challenges for effective treatment. Hispidulin, a flavonoid compound, has shown promising biological effects, particularly in the field of anticancer research. The main objective of this study is to investigate the anticancer properties of hispidulin and gain insight into its mechanistic targets in cancer cells. A comprehensive literature review was conducted to collect data on the anticancer effects of hispidulin. In vitro and in vivo studies were analyzed to identify the molecular targets and underlying mechanisms through which hispidulin exerts its anticancer activities. Hispidulin has shown significant effects on various aspects of cancer, including cell growth, proliferation, cell cycle regulation, angiogenesis, metastasis, and apoptosis. It has been observed to target both extrinsic and intrinsic apoptotic pathways, regulate cell cycle arrest, and modulate cancer progression pathways. The existing literature highlights the potential of hispidulin as a potent anticancer agent. Hispidulin exhibits promising potential as a therapeutic agent for cancer treatment. Its ability to induce apoptosis and modulate key molecular targets involved in cancer progression makes it a valuable candidate for further investigation. Additional pharmacological studies are needed to fully understand the specific targets and signaling pathways influenced by hispidulin in different types of cancer. Further research will contribute to the successful translation of hispidulin into clinical settings, allowing its utilization in conventional and advanced cancer therapies with improved therapeutic outcomes and reduced side effects.

A single-step high-throughput bioassay for quantifying Fc-containing recombinant proteins based on non-classical calculation of fluorescence polarization.

The titer of recombinant proteins is one of the key parameters in biopharmaceutical manufacturing processes. The fluorescence polarization (FP)-based assay, a homogeneous, high-throughput and real-time analytical method, had emerged as a powerful tool for biochemical analysis and environmental monitoring. In this study, an FP-based bioassay was utilized to quantify antibody fragment crystallizable (Fc)-containing proteins, such as recombinant monoclonal antibodies (mAbs) and mAb derivatives, in the cell culture supernatant, and the impacts of tracer molecular weight and FITC-coupling conditions on fluorescence polarization were methodically examined. Distinct from the fluorescence polarization potency calculated by classical formula, we for the first time proposed a new concept and calculation of fluorescence polarization intensity, based on which an analytical method with broader detection range and analysis window was established for quantifying Fc-containing proteins. This provided new ideas for the practical application of fluorescence polarization theory. The established method could detect 96 samples within 30 minutes, with dynamic titer range of 2.5-400 mg L-1, and a linear fitting R2 between the measured and actual concentration reaching 0.99. The method had great application prospects in determining the titer of recombinant proteins with Fc fragments, especially when applied to large-scale screening of high-yield and stable expression CHO cell lines commonly used in biopharmaceutical industry.