Hyaluronic acid/alginate-based biomimetic hydrogel membranes for accelerated diabetic wound repair.

The study aims to develop a new multifunctional biopolymer-based hydrogel membrane dressing by adopting a solvent casting method for the controlled release of cefotaxime sodium at the wound site. Sodium alginate enhances collagen production in the skin, which provides tensile strength to healing tissue. Moreover, the significance of extracellular molecules such as hyaluronic acid in the wound the healing cascade renders these biopolymers an essential ingredient for the fabrication of hydrogel membranes via physical crosslinking (hydrogen bonding). These membranes were further investigated in terms of their structure, and surface morphology, as well as cell viability analysis. A membrane with the most suitable characteristics was chosen as a candidate for cefotaxime sodium loading and in vivo analysis. Results show that the 3D porous nature of developed membranes allows optimum water vapor and oxygen transmission (>8.21 mg/mL) to divert excessive wound exudate away from the diabetic wound bed, MTT assay confirmed cell viability at more than 80%. In vivo results confirmed that the CTX-HA-Alg-PVA hydrogel group showed rapid wound healing with accelerated re-epithelization and a decreased inflammatory response. Conclusively, these findings indicate that CTX-HA-Alg-PVA hydrogel membranes exhibit a suitable niche for use as dressing membranes for healing of diabetic wounds.

Hermetic storage of okra seed maintains seed longevity under changing environment.

Okra seed is vulnerable to loss of germination and vigor in variable storage conditions. High seed moisture contents (SMC) accelerate seed deterioration during storage thus keeping low seed moisture contents by storing seed in hermetic bags may help to retain seed longevity. Okra seed was equilibrated to four initial moisture levels including 8,10, 12 and 14% SMC. Seed was then packed and stored in traditional storage bags (Paper, cloth, polypropylene and jute bag) and hermetic Super Bag for 12 months under ambient conditions. Seed stored in hermetic Super Bag at 8 and 10% moisture contents maintained higher germination due to low seed moisture contents. Moreover, activities of α-amylases and total soluble sugars were higher while electrical conductivity of seed leachates, malondialdehyde (MDA) and reducing sugar contents were less in the seeds stored in hermetic Super Bag at 8 and 10% SMC as compared to seed stored in traditional storage bags. Hermetic storage at 14% moisture negatively influenced the seed quality. Moisture adsorption isotherms of okra seeds were developed at constant temperature of 25°C and varying levels of relative humidity from 60 to 90%. Moisture isotherms indicated no significant increase in seed moisture contents at 60 and 70% relative humidity (RH) in hermetic bags whereas a minor increase in seed moisture at 80 and 90% RH has been observed for the seeds incubated in hermetic bags. SMC significantly increased in traditional storage bags particularly in jute bag at high RH. In conclusion, storage in hermetic bags, maintain low seed moisture and high seed quality. Okra seed storage in hermetic bags at 8 and 10% SMC maintains seed longevity under ambient storage conditions.

Silymarin nanocrystals-laden chondroitin sulphate-based thermoreversible hydrogels; A promising approach for bioavailability enhancement.

Hydrogels has gained tremendous interest as a controlled release drug delivery. However, currently it is a big challenge to attain high drug-loading as well as stable and sustained release of hydrophobic drugs. The poor aqueous solubility and low bioavailability of many drugs have driven the need for research in new formulations. This manuscript hypothesized that incorporation of nanocrystals of hydrophobic drug, such as silymarin into thermoreversible hydrogel could be a solution to these problems. Herein, we prepared nanocrystals of silymarin by antisolvent precipitation technique and characterized for morphology, particle size, polydispersity index (PDI) and zeta potential. Moreover, physical cross-linking of hydrogel formulations based on chondroitin sulphate (CS), kappa-Carrageenan (κ-Cr) and Pluronic® F127 was confirmed by Fourier transformed infrared spectroscopy (FT-IR). The hydrogel gelation time and temperature of optimized hydrogel was 14 ± 3.2 s and 34 ± 0.6 °C, respectively. The release data revealed controlled release of silymarin up to 48 h and in-vivo pharmacokinetic profiling was done in rabbits and further analyzed by high-performance liquid chromatography (HPLC). It is believed that the nanocrystals loaded thermoreversible injectable hydrogel system fabricated in this study provides high drug loading as well as controlled and stable release of hydrophobic drug for extended period.

Bioactive and multifunctional keratin-pullulan based hydrogel membranes facilitate re-epithelization in diabetic model.

Hydrogel membrane dressings with multifunctional tunable properties encompassing biocompatibility, anti-bacterial, oxygen permeability, and adequate mechanical strength are highly preferred for wound healing. The present study aimed to develop biopolymer-based hydrogel membranes for the controlled release of therapeutic agent at the wound site. Toward this end we developed Cefotaxime sodium (CTX) loaded keratin (KR)-pullulan (PL) based hydrogel membrane dressings. All membranes show optimized vapor transmission rate (≥1000 g/ m2/day), oxygen permeability >8.2 mg/mL, MTT confirmed good biocompatibility and sufficient tensile strength (17.53 ± 1.9) for being used as a wound dressing. Nonetheless, KR-PL-PVA membranes show controlled CTX release due to enriched hydrophilic moieties which protect the wound from getting infected. In vivo results depict that CTX-KR-PL-PVA membrane group shows a rapid wound closure rate (p < 0.05) with appreciable angiogenesis, accelerated re-epithelization, and excessive collagen deposition at the wound site. These results endorsed that CTX-KR-PL-PVA hydrogel membranes are potential candidates for being used as dressing material in the diabetic wound.

Self-crosslinked chitosan/κ-carrageenan-based biomimetic membranes to combat diabetic burn wound infections.

Diabetic wound infection often leads to compromised healing with frequent chances of sepsis, amputation and even death. Traditional patient care emphasized on early debridement and fluid resuscitation followed by intravenous antibiotics therapy. However, compromised vasculature often limit the systemic effect of antibiotics. Current study focused formulation of chitosan HCl, κ- carrageenan and PVA based physical cross-linked hydrogel membrane dressings loaded with cefotaxime sodium (CTX), for potential diabetic burn wound healing by adopting solvent casting method. Results of mechanical strength shows tensile strength and % elongation of 12.63 ± 0.25 and 48 ±3.05 respectively. Water vapor transmission rate (WVTR) depicts that despite of formulation KCP3 and KCP6, all hydrogel membranes have WVTR value in range of ideal dressing i.e., 2000-2500 g/m2/day. Whereas, all hydrogel membranes have oxygen permibility values more than 8.2 mg/ml. Bacterial penetration analysis confirms the barrier property of formulated membranes. Drug loaded hydrogel membrane showed control release up to 24 hr which provide protection against bacterial proliferation. Present study aims to constructs diabetic burn rat model which demonstrate that CTX loaded hydrogel membrane shown significantly rapid wound closure higher re-epithelization and numerous granulation tissue formation as compared to positive and negative control group. Conclusively, it is confirmed that formulated hydrogel membranes are beneficial and can be considered as a promising membrane dressing to treat diabetic burn wound.

Lipid A-Ara4N as an alternate pathway for (colistin) resistance in Klebsiella pneumonia isolates in Pakistan.

OBJECTIVES: This study aimed to explore mechanism of colistin resistance amongst Klebsiella pneumoniae isolates through plasmid mediated mcr-1 gene in Pakistan. Carbapenem and Colistin resistant K. pneumoniae isolates (n = 34) stored at - 80 °C as part of the Aga Khan University Clinical Laboratory strain bank were randomly selected and subjected to mcr-1 gene PCR. To investigate mechanisms of resistance, other than plasmid mediated mcr-1 gene, whole genome sequencing was performed on 8 clinical isolates, including 6 with colistin resistance (MIC > 4 µg/ml) and 2 with intermediate resistance to colistin (MIC > 2 µg/ml). RESULTS: RT-PCR conducted revealed absence of mcr-1 gene in all isolates tested. Whole genome sequencing results revealed modifications in Lipid A-Ara4N pathway. Modifications in Lipid A-Ara4N pathway were detected in ArnA_ DH/FT, UgdH, ArnC and ArnT genes. Mutation in ArnA_ DH/FT gene were detected in S3, S5, S6 and S7 isolates. UgdH gene modifications were found in all isolates except S3, mutations in ArnC were present in all except S1, S2 and S8 and ArnT were detected in all except S4 and S7. In the absence of known mutations linked with colistin resistance, lipid pathway modifications may possibly explain the phenotype resistance to colistin, but this needs further exploration.

A TOP6BL mutation abolishes meiotic DNA double-strand break formation and causes human infertility.

Meiosis is pivotal for sexual reproduction and fertility. Meiotic programmed DNA double-strand breaks (DSBs) initiate homologous recombination, ensuring faithful chromosome segregation and generation of gametes. However, few studies have focused on meiotic DSB formation in human reproduction. Here, we report four infertile siblings born to a consanguineous marriage, with three brothers suffering from non-obstructive azoospermia and one sister suffering from unexplained infertility with normal menstrual cycles and normal ovary sizes with follicular activity. An autosomal recessive mutation in TOP6BL was found co-segregating with infertility in this family. Investigation of one male patient revealed failure in programmed meiotic DSB formation and meiotic arrest prior to pachytene stage of prophase I. Mouse models carrying similar mutations to that in patients recapitulated the spermatogenic abnormalities of the patient. Pathogenicity of the mutation in the female patient was supported by observations in mice that meiotic programmed DSBs failed to form in mutant oocytes and oocyte maturation failure due to absence of meiotic recombination. Our study thus illustrates the phenotypical characteristics and the genotype-phenotype correlations of meiotic DSB formation failure in humans.

Gynaecological Cancer Diagnostics: 99mTc-Cisplatin Complex as a Future Approach for Early, Prompt and Efficient Diagnosis of Gynaecological Cancer.

BACKGROUND: Gynaecological cancers (GCCa) are common and have a significant mortality rate all over the world. Early diagnosis of cancer can play a key role in the treatment and survival of a patient. Identification, staging, treatment, and monitoring of gynaecological malignancies is being done successfully by nuclear medicines. DISCUSSION: Currently, single-photon emission computed tomography (SPECT) and positron emission tomography (PET) centered imaging techniques are being developed for use in patients with GCCa as a diagnostic tool. The present work elucidates several clinical studies on the use of radiopharmaceuticals, based on their effectiveness, in the early detection and management of GCCa. It also highlights the importance of reconsidering the biology for nuclear imaging as a future modality for early, rapid and efficient diagnosis of gynecological cancers. This comprehensive review is a part of our study designed to detect gynaecological cancers at an early stage using radionuclide complex, 99m Tc-Cisplatin. CONCLUSION: This article summarizes the significance of radioscintigraphy such as single-photon emission computed tomography (SPECT) and PET for identification of GCCa in the experimental humans and animals.

The Established Nuclear Medicine Modalities for Imaging of Bone Metastases.

BACKGROUND: The skeleton is one of the frequent site of metastases in advanced cancer. Prostate, breast and renal cancers mostly metastasize to bone. DISCUSSION: Malignant tumors lead to significant morbidity and mortality. Identification of bone lesions is a crucial step in diagnosis of disease at early stage, monitoring of disease progression and evaluation of therapy. Diagnosis of cancer metastases is based on uptake of bone-targeted radioactive tracer at different bone remodeling sites. CONCLUSION: This manuscript summarizes already established and evolving nuclear medicine modalities (e.g. bone scan, SPECT, SPECT/CT, PET, PET/CT) for imaging of bone metastases.

Whole exome sequencing identifies a novel dominant missense mutation underlying leukonychia in a Pakistani family.

Hereditary leukonychia (also known as porcelain nails or white nails) is a genetic disorder. It may exist as an isolated feature or associated with other cutaneous or systemic disorders. Although a number of genes have been described to cause leukonychia, still the underlying genetic etiologies of many cases remain unknown. Here, we report a Pakistani family presenting leukonychia and koilonychia nails in mother and five of her kids. All the affected individuals had white to pale nails in appearance exhibiting complete and partial leukonychia, respectively. Similarly, nails of finger and toe appeared brittle and concave, showing the characteristics features of koilonychia. Whole exome sequencing and subsequent Sanger sequencing identified a pathogenic novel missense mutation (c.1390G>A, p.Glu464Lys) in PLCD1, co-segregating with the disorder in an autosomal dominant pattern. In silico prediction tools supported the pathogenicity of the identified mutation. Literature review determined that mutations in PLCD1 only cause leukonychia. Therefore, our findings add another pathogenic variant to the PLCD1 mutation pool causing leukonychia that would help to understand the underlying molecular mechanism.

The evolutionarily conserved genes: Tex37, Ccdc73, Prss55 and Nxt2 are dispensable for fertility in mice.

There are more than 2300 genes that are predominantly expressed in mouse testes. The role of hundreds of these genes has been studied in mouse spermatogenesis but still there are many genes whose function is unknown. Gene knockout (KO) strategy in mice is widely used for in vivo study of gene function. The present study was designed to explore the function of the four genes: Tex37, Ccdc73, Prss55 and Nxt2, which were evolutionarily conserved in eutherians. We found that these genes had a testis-enriched expression pattern in mice except Nxt2. We knocked out these genes by CRISPR/Cas9 individually and found that all the KO mice had normal fertility with no detectable difference in testis/body weight ratios, epididymal sperm counts, as well as testicular and epididymal histology from wild type mice. Although these genes are evolutionarily conserved in eutherians including human and mouse, they are not individually essential for spermatogenesis, testis development and male fertility in mice in laboratory conditions. Our report of these fertile KO data could avoid the repetition and duplication of efforts which will help in prioritizing efforts to focus on genes that are indispensable for male reproduction.

Histone acetyltransferase KAT8 is essential for mouse oocyte development by regulating reactive oxygen species levels.

Proper oocyte development is crucial for female fertility and requires timely and accurate control of gene expression. K (lysine) acetyltransferase 8 (KAT8), an important component of the X chromosome dosage compensation system in Drosophila, regulates gene activity by acetylating histone H4 preferentially at lysine 16. To explore the function of KAT8 during mouse oocyte development, we crossed Kat8flox/flox mice with Gdf9-Cre mice to specifically delete Kat8 in oocytes. Oocyte Kat8 deletion resulted in female infertility, with follicle development failure in the secondary and preantral follicle stages. RNA-seq analysis revealed that Kat8 deficiency in oocytes results in significant downregulation of antioxidant genes, with a consequent increase in reactive oxygen species. Intraperitoneal injection of the antioxidant N-acetylcysteine rescued defective follicle and oocyte development resulting from Kat8 deficiency. Chromatin immunoprecipitation assays indicated that KAT8 regulates antioxidant gene expression by direct binding to promoter regions. Taken together, our findings demonstrate that KAT8 is essential for female fertility by regulating antioxidant gene expression and identify KAT8 as the first histone acetyltransferase with an essential function in oogenesis.

Prevalence and pattern of traumatic limb amputations in female population of Bhimber District, Azad Jammu and Kashmir, Pakistan.

OBJECTIVE: Traumatic limb amputations (TLA) are structural defects that cause mild-to-severe disabilities and have significant impact on the quality of life of subjects. A recent epidemiological study in Bhimber District, AJK, Pakistan, revealed that TLA had a very high incidence among the young/adult females. The present study aimed at determining the prevalence, pattern, causes and risk factors associated with TLA in that female sample. METHODS: In a prospective door-to-door survey, 1731 females were randomly enrolled, and clinico-etiological investigations on 19 subjects with TLA were carried out in a follow-up study. Affected subjects were physically examined and phenotypic detail and restriction to normal function were documented. RESULTS: There were 19 subjects with TLA, hence the prevalence was estimated to be 10.98/1,000 (0.011%; CI=0.0061-0.0159). TLA mostly involved the upper limbs and right hand. Transphalangeal amputations were most frequent, followed by involvements of middle/upper arm and leg segments. These analyses further revealed that agriculture tools were the leading cause of trauma. TLA were witnessed to be higher in subjects originating from Samahni tehsil (OR:2.71), rural areas (OR:3.33), those who were illiterate (OR:2.65), married, spoke Pahari language (OR:1.73), those who fall in higher age category (OR:16.74), and had certain professions. CONCLUSION: Limb amputations had heavy bearings on the lives of subjects. Curiously, majority of these traumas could be avoided by adopting certain safety measures. Prevalence and risk estimate of TLA across various socio-demographic variables of Bhimber population would be useful in guiding further studies and the public health policy to mitigate the impact of these anomalies.

Consanguinity and its sociodemographic differentials in Bhimber District, Azad Jammu and Kashmir, Pakistan.

Kashmiri population in the northeast of Pakistan has strong historical, cultural and linguistic affinities with the neighbouring populations of upper Punjab and Potohar region of Pakistan. However, the study of consanguineous unions, which are customarily practised in many populations of Pakistan, revealed marked differences between the Kashmiris and other populations of northern Pakistan with respect to the distribution of marriage types and inbreeding coefficient (F). The current descriptive epidemiological study carried out in Bhimber district of Mirpur division, Azad Jammu and Kashmir, Pakistan, demonstrated that consanguineous marriages were 62% of the total marriages (F=0.0348). First-cousin unions were the predominant type of marriages and constituted 50.13% of total marital unions. The estimates of inbreeding coefficient were higher in the literate subjects, and consanguinity was witnessed to be rising with increasing literacy level. Additionally, consanguinity was observed to be associated with ethnicity, family structure, language, and marriage arrangements. Based upon these data, a distinct sociobiological structure, with increased stratification and higher genomic homozygosity, is expected for this Kashmiri population. In this communication, we present detailed distribution of the types of marital unions and the incidences of consanguinity and inbreeding coefficient (F) across various sociodemographic strata of Bhimber/Mirpuri population. The results of this study would have implication not only for other endogamous populations of Pakistan but also for the sizeable Kashmiri community immigrated to Europe.

Prevalence of Congenital Anomalies and Non-Communicable Diseases in Women of Age 12-75 Years in District Bhimber, Azad Jammu and Kashmir, Pakistan.

BACKGROUND: The advancement in the healthcare systems, stringent interventions for infectious diseases and improved diet has significantly shifted the patterns of morbidities, and consequently hereditary and congenital anomalies (CA) and non-communicable diseases (NCDs) have emerged as the most common causes of morbidity and mortality. In Pakistan, there is no systematic health surveillance system to assess the impact of such diseases particularly on the young and adult populations. METHODS: In order to glean into the health and morbidity profile of Azad Jammu and Kashmir we have carried out an epidemiological study in Bhimber District in the north-east of Pakistan. A total of 1,731 female subjects of age 12-75 yr originating from Bhimber were recruited through a cross-sectional study. RESULTS: There were 74 cases (and 15 types) of CA with a prevalence estimate of 42.75/1,000. CA was significantly higher in subjects who were illiterate and married, speaking Pahari language and belonged to rural areas and nuclear families. Additionally, there were 104 cases (and 21 types) of NCDs (prevalence 60.08/1,000). NCDs were observed to have higher prevalence in subjects who were illiterate and married, speaking Punjabi language, and belonged to higher age groups and nuclear families. CONCLUSION: This study explores the types and dynamics of morbidity across the major socio-demographic parameters of adult females of Bhimber and would be helpful in estimating the impact of morbidity in this population. A comprehensive country-wide study is the need of the time to identify specific risk factors associated with certain morbidity types and help prioritize areas for interventions.

Zygodactyly with thumb aplasia: an unusual variant in a male subject.

This is a presentation of a male subject with unusual combination of limb malformations. The subject had unilateral zygodactyly of the left foot with thumb aplasia in the right hand. Further, the webbing between second and third toes was complete culminating in osseous fusion of the terminal phalanges and valgus deviation of the affected digits of the foot. The nails were also involved but had separate origins. In the right hand, first digital ray was observed to be completely omitted. There was aplasia of certain carpals while the radius showed minimal clinical symptoms. The subject was the product of first cousins union. To the best of our knowledge, this combination of limb phenotype has not been described before.