Transfer to the clinic: refining forward programming of hPSCs to megakaryocytes for platelet production in bioreactors.

The production of in vitro-derived platelets has great potential for transfusion medicine. Here, we build on our experience in the forward programming (FoP) of human pluripotent stem cells (hPSCs) to megakaryocytes (MKs) and address several aspects of the complex challenges to bring this technology to the bedside. We first identify clinical-grade hPSC lines that generate MKs efficiently. We design a bespoke media to maximize both production and maturity of MKs and improve platelet output. Crucially, we transition the lentiviral-based FoP of hPSCs to a nonviral inducible system. We also show how small molecules promote a definitive hematopoiesis phenotype during the differentiation process, thereby increasing the quality of the final product. Finally, we generate platelets using a bioreactor designed to reproduce the physical cues that promote platelet production in the bone marrow. We show that these platelets are able to contribute to both thrombus formation in vitro and have a hemostatic effect in thrombocytopenic mice in vivo.

Frequency and trend of vitamin D deficiency in correlation with demographic and selected biochemical parameters in Karachi, Pakistan.

OBJECTIVE: To evaluate the frequency and trend of vitamin D deficiency, and to find its correlation with demographic and selected biochemical parameters. METHODS: The retrospective study was conducted at The Indus Hospital, Karachi, and comprised clinical laboratory data of individuals tested for vitamin D from January 2013 to March 2018. The trend of vitamin D deficiency and frequency was assessed in relation to age, gender and serum levels of calcium, phosphorus, magnesium, alkaline phosphatase and parathyroid hormone. Data was analysed using Stata software. RESULTS: Of the 35,017 tests analysed, 23,522 (67.2%) related to females and 11,495 (32.8%) to males (p<0.05). Overall, 25,051 (71.5%) were vitamin D-deficient while 504 (1.4%) had toxic levels. Age had significant correlation with vitamin D deficiency (p<0.05). No significant correlation was observed with any of the biochemical parameters studied (p>0.05). CONCLUSIONS: Vitamin D deficiency was high among female gender and young population.

Frequency of isolation and susceptibility pattern of E. coli from in and outpatients over a period of five years at The Indus Hospital Network, Karachi Pakistan.

OBJECTIVE: To determine the frequency of isolation and susceptibility pattern of Escherichia coli at a urban health centre. METHODS: The retrospective study was conducted at the Indus Hospital Network, Karachi, and comprised Enterobacteriaceae isolates obtained from urine, blood and routine culture from patients presenting between 2013 and 2017. The samples were evaluated for resistance patterns against a range of antibiotics and frequency of isolation was determined. Data was analysed using SPSS version 21.0. RESULTS: Of the 10,667 isolates analysed, 6380(60%) were Escherichia coli. Outpatient and inpatient isolates constituted 4184(65.6%) and 2196(34.4%) of the total E. coli isolates respectively. Of the 1446(22.66%) isolates obtained from urine, 1007(59.64%) had E. coli which was the highest isolation rate among all types of samples. There was a significant difference in resistance rates between inpatient and outpatient isolates for first-line and second-line injectable and oral antibiotics (p<0.05). CONCLUSIONS: Escherichia coli isolates showed high resistance towards co-amoxiclav, ampicillin, ceftriaxone and ciprofloxacin, leaving little empirical options for treating outpatients.

8p11 associated myeloid/lymphoid neoplasms: An illustrative case report of this rare entity and review of literature.

This report describes a unique case of 8p11 myeloproliferative syndrome (EMS), also known as stem cell leukaemia-lymphoma syndrome. A 13 years old male was referred from a tertiary care hospital after cervical lymph node biopsy. The disease mechanism of this neoplasm is to either evolve into acute myeloid leukemia or mixed lineage leukaemia, and less frequently of T or B lymphoid lineage. However, here we show a case of a rare simultaneous presentation of T lymphoblastic lymphoma on cervical lymph node and B lymphoblastic leukaemia on bone marrow biopsy along with t (8; 13) on karyotype testing.

Minimal residual disease in childhood B Lymphoblastic Leukemia and its correlation with other risk factors.

OBJECTIVE: To determine frequency of post induction and post consolidation minimal residual disease (MRD) in pediatric B-lymphoblastic leukemia (B-ALL) patients and its association with clinical risk factors. METHODS: This is a retrospective, cross sectional study carried out at the Indus Hospital on paediatric patients (1-17 years) was performed from May 2015 to January 2018. On day 35, MRD testing was done on bone marrow aspirate using four color flow cytometer with 0.01% cut off. Positive cases were retested at post consolidation. Data was collected for demographics, total leukocyte count (TLC), central nervous system status (CNS), Cytogenetics for BCR-ABL, MLL, TEL-AML by FISH and prophase response then analyzed in association to MRD status. RESULTS: Out of 362 patients, 133 (37%) were post induction MRD positive, with no statistically significant association to age, gender, TLC, CNS status, prophase response, BCR-ABL and TEL-AML1. However, MLL showed closely significant association (p-value=0.05). Post consolidation, 49 (44%) were MRD positive; age, National cancer institute (NCI) risk groups and CNS status showed statistical significance (p-value <0.05). CONCLUSION: Despite high frequency of MRD positivity, significant association is not observed between post induction MRD and risk factors. However, post consolidation MRD has a significant association with NCI risk groups, age and CNS status.