Current stage of preclinical and clinical development of guggulsterone in cancers: Challenges and promises.

Throughout human history, the utilization of medicinal herbs has been recognized as a crucial defense against various ailments, including cancer. Natural products with potential anticancer properties, capable of inducing apoptosis in cancer cells, have garnered substantial attention. One such agent under investigation is guggulsterone (GS), a phytosterol derived from the gum resin of the Commiphora mukul tree. This review aims to provide a comprehensive summary of recent studies elucidating the anticancer molecular mechanisms and molecular targets of GS, guiding future research and potential applications as an adjuvant drug in cancer therapy. Recent in vivo and in vitro studies have explored the biological activities of the active ingredients in Commiphora mukul. Specifically, GS emerges as a potential cancer chemopreventive and therapeutic agent. The investigations delve into the impact of GS on constitutively activated survival pathways, including Janus kinase/signal transducer and activator of transcription (JAK/STAT), nuclear factor-kappa B (NF-kB), and PI3-kinase/AKT signaling pathways. These pathways regulate antiapoptotic and proinflammatory genes, exerting control over growth and inflammatory responses. The findings highlight the potential of GS in disrupting survival pathways crucial for cancer cell viability. The inhibition of JAK/STAT, NF-kB, and PI3-kinase/AKT signaling pathways positions GS as a promising candidate for cancer therapy. The review synthesizes evidence from diverse studies, underscoring the multifaceted biological activities of GS in cancer prevention and treatment. To advance our understanding, future clinical and translational studies are imperative to determine effective doses in humans. Additionally, there is a need for the development of new pharmaceutical forms of GS to optimize therapeutic effects. This comprehensive review provides a foundation for ongoing research, offering insights into the potential of GS as a valuable addition to the armamentarium against cancer.

Morphological and Molecular Characterizations of Arbuscular Mycorrhizal Fungi and Their Influence on Soil Physicochemical Properties and Plant Nutrition.

Pulses are considered a remarkable and stable source of nutrients, which are being presently extensively cultivated and consumed in different parts of the world. Pulses belong to the family Leguminosae and are a rich source of nutrients such as phosphorus (P) and nitrogen (N) for best growth via symbiotic relationship with bacteria and AMF (arbuscular mycorrhizal fungi). The aim of the current study was evaluating the influence of AMF diversity associated with various pulses (French bean, mung bean, kidney bean, peas, soybean, peanuts, and grams). Furthermore, AMF characterization was done using morphological features of spores and sequencing of the rDNA gene, which confirmed the existence of 10 different AMF taxa. Among the different genera, the genus Glomus was observed to be the most dominant with 30% species followed by Gigaspora (22%), Sclerocystis (12%), Acaulospora (8%), Rhizophagus and Septoglomus (7%), Diversispora (5%), and Claroideoglomus, Archaeospora, and Ambispora (3%). Furthermore, soil physicochemical analysis and percentage of AMF colonization results revealed the fact that the phosphorus content (inversely proportional to the AMF diversity) was a determining factor of AMF diversity. The highest amount of available phosphorus (62.825 mg kg-1) in the district Swabi resulted in a low rate of AMF colonization (6.66 ± 11.54%) with a comparatively higher rate of AMF colonization (50.66 ± 1.15%) found in the soil of the district Chitral having a low phosphorus content (17.3 ± 7.6 mg kg-1). Nutrient uptake by pulses including nitrogen (2.4 ± 1.3%), phosphorus (13.5 ± 7.6 mg kg -1), potassium (99.5 ± 25.8 mg kg -1), zinc (1.4 ± 0.5 mg kg -1), moisture (2.3 ± 1.3%), crude fats (5.6 ± 2.8%), ash (4 ± 1.2%), and proteins (13.6 ± 9.01%) determined the fact that AMF species diversity is positively correlated to the plant mineral nutrition. From the current study, it is concluded that AMF inoculation to the soil fields is beneficial to ensure the sustainability and productivity of pulse crops in diverse environmental conditions without polluting the soil.

Rosmarinic acid and its derivatives: Current insights on anticancer potential and other biomedical applications.

Cancer is still the leading cause of death worldwide, burdening the global medical system. Rosmarinic acid (RA) is among the first secondary metabolites discovered and it is a bioactive compound identified in plants such as Boraginaceae and Nepetoideae subfamilies of the Lamiaceae family, including Thymus masticmasti chinaythia koreana, Ocimum sanctum, and Hyptis pectinate. This updated review is to highlight the chemopreventive and chemotherapeutic effects of RA and its derivatives, thus providing valuable clues for the potential development of some complementary drugs in the treatment of cancers. Relevant information about RA's chemopreventive and chemotherapeutic effects and its derivatives were collected from electronic scientific databases, such as PubMed/Medline, Scopus, TRIP database, Web of Science, and Science Direct. The results of the studies showed numerous significant biological effects such as antiviral, antibacterial, anti-inflammatory, anti-tumour, antioxidant and antiangiogenic effects. Most of the studies on the anticancer potential with the corresponding mechanisms are still in the experimental preclinical stage and are missing evidence from clinical trials to support the research. To open new anticancer therapeutic perspectives of RA and its derivatives, future clinical studies must elucidate the molecular mechanisms and targets of action in more detail, the human toxic potential and adverse effects.

A comprehensive review on traditional uses, phytochemistry and pharmacological properties of Paeonia emodi Wall. ex Royle: current landscape and future perspectives.

Paeonia emodi Wall. ex Royle is commonly known as Himalayan paeony has great importance as a food and medicine. The practice of Paeonia emodi Wall. ex Royle is very ancient and it is conventionally used for a wide range of illnesses in the folk system of medicine because of its wide beneficial phytochemical profile. The main purpose of the current review was the synthesis of recent data on botany, ethnopharmacology, phytochemistry and potential pharmacological mechanisms of action of Paeonia emodi Wall. ex Royle, thus offering new prospects for the development of new adjuvant natural therapies. Using scientific databases such as PubMed/MedLine, Scopus, Web of Science, ScienceDirect, Google Scholar, Springer, and Wiley, a comprehensive literature search was performed for Paeonia emodi Wall. ex Royle. For searching, we used the next MeSH terms: "Biological Product/isolation and purification", "Biological Products/pharmacology", "Drug Discovery/methods", "Ethnopharmacology, Medicine", "Traditional/methods", "Paeonia/chemistry", "Plant Extracts/pharmacology", "Phytochemicals/chemistry", "Phytochemicals/pharmacology", "Plants, Medicinal". The results of the most recent studies were analyzed and the most important data were summarized in tables and figures. Phytochemical research of Paeonia emodi Wall. ex Royle has led to the isolation of triterpenes, monoterpenes, phenolic acids, fatty acids, organic compounds, steroids, free radicals and some other classes of primary metabolites. In addition, diverse pharmacological activities like antibacterial, antifungal, anticoagulant, airway relaxant lipoxygenase and beta-glucuronidase inhibiting activity, radical scavenging activity, phytotoxic and insecticidal activities have been reported for Paeonia emodi Wall. ex Royle. Different bioactive compounds of Paeonia emodi Wall. ex Royle has proven their therapeutic potential in modern pharmacological and biomedical research to cure numerous gastrointestinal and nervous disorders. In future, further in vitro and in vivo therapeutic studies are required to identify new mechanisms of action, pharmacokinetics studies, and new pharmaceutical formulations for target transport and possible interaction with allopathic drugs. Also, new research regarding quality evaluation, toxicity and safety data in humans is needed.

Exploring Physical Characterization and Different Bio-Applications of Elaeagnus angustifolia Orchestrated Nickel Oxide Nanoparticles.

Elaeagnus angustifolia (EA) mediated green chemistry route was used for the biofabrication of NiONPs without the provision of additional surfactants and capping agents. The formation of NiONPs was confirmed using advanced different characterization techniques such as Scanning electron microscopy, UV, Fourier transmission-infrared, RAMAN, and energy dispersal spectroscopic and dynamic light scattering techniques. Further, different biological activities of EA-NiONPs were studied. Antibacterial activities were performed using five different bacterial strains using disc-diffusion assays and have shown significant results as compared to standard Oxytetracycline discs. Further, NiONPs exhibited excellent antifungal performance against different pathogenic fungal strains. The biocompatibility test was performed using human RBCs, which further confirmed that NiONPs are more biocompatible at the concentration of 7.51-31.25 µg/mL. The antioxidant activities of NiONPs were investigated using DPPH free radical scavenging assay. The NiONPs were demonstrated to have much better antioxidant potentials in terms of % DPPH scavenging (93.5%) and total antioxidant capacity (81%). Anticancer activity was also performed using HUH7 and HEP-G2 cancer cell lines and has shown significant potential with IC50 values of 18.45 µg/mL and 14.84 µg/mL, respectively. Further, the NiONPs were evaluated against Lesihmania tropica parasites and have shown strong antileishmanial potentials. The EA-NiONPs also showed excellent enzyme inhibition activities; protein kinase (19.4 mm) and alpha-amylase (51%). In conclusion, NiONPs have shown significant results against different biological assays. In the future, we suggest various in vivo activities for EA-NiONPs using different animal models to further unveil the biological and biomedical potentials.

Rhamnella gilgitica functionalized green synthesis of ZnONPs and their multiple therapeutic properties.

In the recent years, green synthesis of zinc oxide nanoparticles (ZnONPs) using plant extracts and phytochemicals has gained significant attention. In present research study, facile, green, and tunable ZnONPs were biosynthesized from Rhamnella gilgitica leaf aqueous extract as a strong reducing and stabilizing agents. The prepared ZnONPs@Rhamnella were characterized and validated using common nanotechnology techniques (UV-Vis, XRD, EDX, FT-IR, SEM, TEM, DLS, and Raman) and revealed spherical morphology with particle size ~21 nm. The asynthesized ZnONPs were further evaluated for different biological applications. Strong antimicrobial efficacies were reported for ZnONPs using disc-diffusion method and were capable of rendering significant antimicrobial potential. ZnONPs were evaluated against HepG2 (IC50 : 18.40 µg/ml) and HUH7 (IC50 : 20.59 µg/ml) cancer cell lines and revealed strong anticancer properties. Dose-dependent MTT cytotoxicity assay was confirmed using Leishmania tropica "KWH23 strain" (promastigote: IC50 : 26.78 µg/ml and amastigote: IC50 : 29.57 µg/ml). Antioxidant activities (DPPH: 93.36%, TAC: 72.43%) were performed to evaluate their antioxidant potentials. Further, protein kinase and α-amylase inhibition assays were determined. Biocompatibility assays were done using human RBCs and macrophages thus revealed biosafe and non-toxic nature of ZnONPs@Rhamnella. In current experiment, we concluded that greenly orchestrated ZnONPs is an attractive, non-toxic and ecofriendly candidate and showed potential biological activities. In future, different clinical trials and in vivo studies are necessary for the confirmation of these remedial properties of ZnONPs using different animal models. RESEARCH HIGHLIGHTS: Greenly orchestrated ZnONPs were synthesized using Rhamnella gilgitica leaves broth. Analytical techniques such as UV, SEM, TEM, XRD, FTIR, DLS, and Raman confirmed synthesis of ZnONPs. Green ZnONPs determined strong antimicrobial, cytotoxic, and antioxidant potentials. Significant enzyme inhibition and biocompatibility assays were investigated.

Chemical characterization and evaluation of the nephroprotective potential of Parrotiopsis jacquemontiana (Decne) Rehder and Periploca hydaspidis Falc crude extract in CCl4-induced Male Sprague-Dawley Rats.

Biochemical, antioxidant, serum, and urine profiles together with physical examination can deliver important information regarding animal health status, and are vital in the diagnosis and treatment of patients. CCl4, a potent nephrotoxin, was used for causing toxicity in rat kidneys. The present study aimed at exploring the nephroprotective potential of P. jacquemontiana leaves methanol extract (PJM) and P. hydaspidis whole-plant methanol extract (PHM) on kidney cells of male rats after oxidative stress and DNA damage was instigated by CCl4. Various parameters including enzymatic levels, serum profiles, urine profiles, genotoxicity, and histological studies were conducted. In renal samples of rats treated with CCl4, the antioxidant enzymes (POD, SOD, CAT), PH level, protein level, and glutathione contents were significantly (p < 0.05) declined whereas renal biochemicals (H2O2, TBARS, and nitrite), specific gravity, level of urea, urobilinogen, serum BUN and creatinine were markedly (p < 0.05) increased relative to control group. Co-administration of PJM and PHM with CCl4 displayed protective ability against CCl4 intoxication by restoring activities of antioxidant enzymes, urine profile, biochemical parameters, and serum profile in rats. CCl4 also induced prominent DNA damages and glomerular atrophy with abnormal appearance of glomerulus and Bowman's capsule. These damages results in impaired corticular sections, edema in Bowman's capsule, accumulation of necrotic cells, dilation of convoluted tubules, and narrowing of space between Bowman's capsule, which were successfully ameliorated after co-administration of PJM and PHM fractions in a dose-dependent manner (200 and 400 mg/kg b.w.). The results obtained suggest the therapeutic role of PJM and PHM in oxidative-stress related disorders of kidney and may be helpful in kidney trauma.

Unfolding molecular switches in plant heat stress resistance: A comprehensive review.

KEY MESSAGE: Plant heat stress response is a multi-factorial trait that is precisely regulated by the complex web of transcription factors from various families that modulate heat stress responsive gene expression. Global warming due to climate change affects plant growth and development throughout its life cycle. Adds to this, the frequent occurrence of heat waves is drastically reducing the global crop yield. Molecular plant scientists can help crop breeders by providing genetic markers associated with stress resistance. Plant heat stress response (HSR), however, is a multi-factorial trait and using a single stress resistance trait might not be ideal to develop thermotolerant crops. Transcription factors participate in regulation of plant biological processes and environmental stress responses. Recent studies have revealed that plant HSR is precisely regulated by the complex web of transcription factors from various families. These transcription factors enhance plant heat stress tolerance by regulating the expression level of several stress-responsive genes independently or in cross talk with different other transcription factors. This review explores how signaling pathways triggered by heat stress are regulated by multiple transcription factor families. To our knowledge, we for the first time analyze the role of major transcription factor families in plant HSR along with their regulatory mechanisms. In the end, we will also discuss the potential of emerging technologies to improve thermotolerance in plants.

Green synthesis of zinc oxide nanoparticles using Elaeagnus angustifolia L. leaf extracts and their multiple in vitro biological applications.

Due to their versatile applications, ZnONPs have been formulated by several approaches, including green chemistry methods. In the current study, convenient and economically viable ZnONPs were produced using Elaeagnus angustifolia (EA) leaf extracts. The phytochemicals from E. angustifolia L. are believed to serve as a non-toxic source of reducing and stabilizing agents. The physical and chemical properties of ZnONPs were investigated employing varying analytical techniques (UV, XRD, FT-IR, EDX, SEM, TEM, DLS and Raman). Strong UV-Vis absorption at 399 nm was observed for green ZnONPs. TEM, SEM and XRD analyses determined the nanoscale size, morphology and crystalline structure of ZnONPs, respectively. The ZnONPs were substantiated by evaluation using HepG2 (IC50: 21.7 µg mL-1) and HUH7 (IC50: 29.8 µg mL-1) cancer cell lines and displayed potential anticancer activities. The MTT cytotoxicity assay was conducted using Leishmania tropica "KWH23" (promastigotes: IC50, 24.9 µg mL-1; and amastigotes: IC50, 32.83 µg mL-1). ZnONPs exhibited excellent antimicrobial potencies against five different bacterial and fungal species via the disc-diffusion method, and their MIC values were calculated. ZnONPs were found to be biocompatible using human erythrocytes and macrophages. Free radical scavenging tests revealed excellent antioxidant activities. Enzyme inhibition assays were performed and revealed excellent potential. These findings suggested that EA@ZnONPs have potential applications and could be used as a promising candidate for clinical development.

Characterization and phytochemical constituents of Periploca hydaspidis Falc crude extract and its anticancer activities.

The current study aims to investigate the anticancer potential of Periploca hydaspidis extracts against HCCLM3 and MDA-MB 231 cell lines with invasive properties and to identify molecular targets underlying its action mechanism. Cytotoxic screening of plant extracts was done via MTT assay against liver and breast cancer cell lines and GC/MS of the best cytotoxic fraction was performed to identify its chemical composition. Flow cytometry detected apoptosis and cell-cycle changes after drug treatment. The specified cells were studied for migration and invasion potential along with performing western blot analysis of proteins involved in apoptosis, cell-cycle, metastasis, and MAPK (Mitogen-activated protein kinase) cell-signaling pathway. The results revealed the crude methanol (PHM) fraction of P. hydaspidis shown dose and time dependent cell-proliferative inhibition response. GC/MS analysis detected 54 compounds of which fatty acids (29.8%), benzenoids (15.7%), and esters (14.3%) constituted the bulk. The inhibitory effect against cancer cells was linked with cell-cycle arrest at G0/G1 phase, induction of apoptosis, reduced migration and invasion capabilities post treatment. PHM induced apoptosis via downregulation of anti-apoptotic (survivin, B-cell lymphoma Extra-large; BCL-XL, X-linked inhibitor of apoptosis protein; XIAP, Myelocytomatosis; C-myc), metastatic (Matrix metallopeptidases 9/2; MMP9/2), and cell-cycle regulatory (cyclin D1 and E) proteins, whereas upregulation of pro-apoptotic proteins (Bcl-2 homologous antagonist/killer; BAK, Bcl-2-Associate X protein; BAX, cleaved caspases; 3,7,8,9, and PARP) and activation of MAPK (Jun amino-terminal kinase; JNK and P38) pathway. P38 was needed for PHM-induced apoptosis, where the inhibition of P38 by pharmacological inhibitor (SB239063) diminished the apoptotic effects. Overall, our results conclude that PHM can inhibit cell-proliferation and induce apoptotic effects by activation of P38 MAPK cell-signaling pathway. This suggests the methanol fraction of P. hydaspidis (PHM) to have anticancer compounds, potentially useful for treating liver and breast cancer. In future, one-step advance studies of PHM regarding its role in metastatic inhibition, immune response modulation for reducing tumor, and inducing apoptosis in suitable animal models would be an interesting and promising research area.

Chemical composition and pharmacological bio-efficacy of Parrotiopsis jacquemontiana (Decne) Rehder for anticancer activity.

Consistent STAT3 (Single transducer and activator of transcription 3) activation is observed in many tumors and promotes malignant cell transformation. In the present investigation, we evaluated the anticancer effects of Parrotiopsis jacquemontiana methanol fraction (PJM) on STAT3 inhibition in HCCLM3 and MDA-MB 231 cells. PJM suppressed the activation of upstream kinases i.e. JAK-1/2 (Janus kinase-1/2), and c-Src (Proto-oncogene tyrosine-protein kinase c-Src), and upregulated the expression levels of PIAS-1/3 (Protein Inhibitor of Activated STATs-1/3), SHP-1/2 (Src-homology region 2 domain-containing phosphatase-1/2), and PTP-1ß (Protein tyrosine phosphatase 1 ß) which negatively regulate STAT3 signaling pathway. PJM also decreased the levels of protein products conferring to various oncogenes, which in turn repressed the proliferation, migration, invasion, and induced apoptosis in cancer cell lines. The growth inhibitory effects of PJM on cell-cycle and metastasis were correlated with decreased expression levels of CyclinD1, CyclinE, MMP-2 (Matrix metalloproteinases-2), and MMP-9 (Matrix metalloproteinases-9). Induction of apoptosis was indicated by the cleavage and subsequent activation of Caspases (Cysteine-dependent Aspartate-directed Proteases) i.e. caspase-3, 7, 8, 9, and PARP (Poly (ADP-ribose) polymerase) as well as through the down-regulation of anti-apoptotic proteins. These apoptotic effects of PJM were preceded by inhibition of STAT3 cell-signaling pathway. STAT3 was needed for PJM-induced apoptosis, and inhibition of STAT3 via pharmacological inhibitor (Stattic; SC-203282) abolished the apoptotic effects. Conclusively, our results demonstrate the capability of PJM to inhibit cancer cell-proliferation and induce apoptosis by suppressing STAT3 via upregulation of STAT3 inhibitors and pro-apoptotic proteins whereas the down-regulation of upstream kinases and anti-apoptotic protein expression. In future, one-step advance studies of PHM regarding its role in metastatic inhibition, immune response modulation for reducing tumor, and inducing apoptosis in suitable animal models would be an interesting and promising research area.

Molecular mechanisms of plant tolerance to heat stress: current landscape and future perspectives.

KEY MESSAGE: We summarize recent studies focusing on the molecular basis of plant heat stress response (HSR), how HSR leads to thermotolerance, and promote plant adaptation to recurring heat stress events. The global crop productivity is facing unprecedented threats due to climate change as high temperature negatively influences plant growth and metabolism. Owing to their sessile nature, plants have developed complex signaling networks which enable them to perceive changes in ambient temperature. This in turn activates a suite of molecular changes that promote plant survival and reproduction under adverse conditions. Deciphering these mechanisms is an important task, as this could facilitate development of molecular markers, which could be ultimately used to breed thermotolerant crop cultivars. In current article, we summarize mechanisms involve in plant heat stress acclimation with special emphasis on advances related to heat stress perception, heat-induced signaling, heat stress-responsive gene expression and thermomemory that promote plant adaptation to short- and long-term-recurring heat-stress events. In the end, we will discuss impact of emerging technologies that could facilitate the development of heat stress-tolerant crop cultivars.

Green synthesis of nickel oxide nanoparticles using leaf extract of Berberis balochistanica: Characterization, and diverse biological applications.

In current report, nickel oxide nanoparticles (NiONPs) were synthesized using leaf extract of Berberis balochistanica (BB) an endemic medicinal plant. The BB leaves extract act as a strong reducing, stabilizing, and capping agent in the synthesis of BB@NiONPs. Further, BB@NiONPs were characterized using Uv-visible spectroscopy (UV-vis), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), Energy dispersive spectroscopy (EDS), scanning electron microscopy (SEM), and average size was calculated ~21.7 nm). Multiple in vitro biological activities were performed to determine their therapeutic potentials. The BB@NiONPs showed strong antioxidant activities in term of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and total antioxidant capacity (TAC) with scavenging potential of 69.98 and 59.59% at 200 µg/ml, respectively. The antibacterial and antifungal testes were examined using different bacterial and fungal strains and dose-dependent inhibition response was reported. Laterally, cytotoxic and phytotoxic activities were studied using brine shrimp and radish seeds. The result determined potential cytotoxic activity with LD50 value (49.10 µg/ml) and outstanding stimulatory effect of BB@NiONPs on seed germination at lower concentrations as compared to control. Overall, result concluded that biosynthesis of NiONPs using leaf extracts of Berberis balochistanica is cheap, easy, and safe method and could be used in biomedical and agriculture field as nanomedicine and nano fertilizer.

Scanning electron microscopy of Sophora alopecuroides L. seeds and their cytotoxic, antimicrobial, antioxidant, and enzyme inhibition potentials.

Sophora alopecuroides L. is a highly medicinal plant. The aim of the current study was to determine the phytochemical screening, pharmacological potentials and application of scanning electron microscope (SEM) of S. alopecuroides (SA) seeds. To achieve this purpose, six different solvents were used to prepare SA seed extracts. Phytochemical and antioxidant activities were determined calorimetrically. To investigate the antidiabetic activity, α-amylase inhibition assay was determined. Brine shrimp assay was used to determine cytotoxicity potential. Anti-leishmanial potential was confirmed using MTT assay. Disc-diffusion method was used to detect protein kinase inhibitory, antibacterial and antifungal activities and showed significant results. SEM analysis was used as an identification tool. Considerable amount of phenolic and flavonoid contents were identified in methanol extract (SASM) (93.76 ± 2.71 GAE/mg) and (77 ± 3.60 QE/mg). Highest DPPH scavenging potential (82%) was reported for SASM. Significant total antioxidant capacity (90.60 ± 1.55 alpha amylase enzyme [AAE]/mg) and total reducing power (94.44 ± 1.38 AAE/mg) were determined for LOSM. Highest α-amylase inhibition was reported in SASM (78.20 ± 1.58%). Highest LD50 of brine shrimp was found for n-hexane extract (SASH) 13.03 µg/ml. All extracts showed strong anti-leishmanial activity except SASH. The seeds of SA were seen to be oblong to obovate, projections, wavy slightly straight, anticlinal wall was raised with apex acuminate. In conclusion, our experimental findings highly support the ethnomedicinal and biological potentials of the SA seeds. Moreover, SA seeds need to be explored for identification and isolation of bioactive compounds. In future, we recommend further in vivo toxicity assays and clinical efficacies to further evaluate its different biomedical properties.

Molecular Characterization of ß-Thalassemia Mutations Via the Amplification Refractory Mutation System-Polymerase Chain Reaction Method at the North Waziristan Agency, Pakistan.

ß-Thalassemia (ß-thal) is a monogenic disease characterized by mutations on the HBB gene, affecting the production of globin that results in hypochromic and microcytic anemia. The aim of this study was to determine the prevalence of six common ß-thal mutations, and their frequency and inheritance pattern in affected populations of North Waziristan Agency, Pakistan. In this study, 130 blood samples from 37 unrelated ß-thalassemic families having a minimum of one transfusion-dependent child with ß-thal major (ß-TM), were retrieved either from the Thalassaemia Centre for Women and Children Hospital Bannu or their home towns situated in Noth Waziristan Agency. All samples were analyzed by the amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) using six allele-specific primers for the presence of the six ß-thal mutations common in the Pakistani population. Of the six common mutations, our study demonstrated five HBB mutations comprising HBB: c.27_28insG, HBB: c.92+5G>C, HBB: c.126_129delCTTT, HBB: c.92+1G>T and HBB: c.17_18delCT from the families studied, while mutation HBB: c.47G>A [codon 15 (G>A)] was not detected in any of the studied families. Furthermore, the HBB: c.27_28insG and HBB: c.92+5G>C were noted to be the most common with frequencies of 42.85 and 31.42%, respectively. The findings of the present study may be useful in launching carrier screening and prenatal diagnosis (PND) programs by screening analyzed and other unanalyzed affected families for the possible presence of common mutations through the ARMS-PCR technique that will help to control the disease.

Investigation of molecular heterogeneity of ß-thalassemia disorder in District Charsadda of Pakistan.

OBJECTIVE: Thalassemia is blood related disease which arises from the reduced level of hemoglobin in red blood cells (RBC), a protein responsible for carrying oxygen inside the body. Considering its widespread occurrence in developing countries like Pakistan, this study aims to investigate the common molecular anomalies of the beta thalassemia disease in district Charsadda, Khyber Pakhtunkhwa. METHODS: This work was done at Abdul Wali Khan University (AWKU) Mardan, Khyber Pakhtunkhwa, Pakistan. The work was performed on the blood samples collected from the patients and their families with beta thalassemia major (n = 13 families) belonged to District Charsadda. The collected blood samples were analyzed for presence of six known mutations with the help of polymerase cha in reaction technique i.e. amplification of refractory mutation system. RESULTS: Our Study reports six known mutations (IVS-1-5, FSC 8/9, CD 41/42, IVS-1-1, CD 15 and FSC-5) accounting for about 90% of total beta thalassemia genes in this country. Among the reported mutations, IVS 1-5 was the most prevalent beta thalassemia gene in patients belonging to District Charsadda. CONCLUSION: The results and findings of the current study may help in accessing the frequency of these common mutations and in initiating pre-natal diagnosis programme in Pakistan.