The covalent transfer of the AMP portion of ATP onto a target protein—termed adenylylation or AMPylation—by the human Fic protein HYPE/FICD has recently garnered attention as a key regulatory mechanism in endoplasmic reticulum homeostasis, neurodegeneration, and neurogenesis. As a central player in such critical cellular events, high-throughput screening (HTS) efforts targeting HYPE-mediated AMPylation warrant investigation. Herein, we present a dual HTS assay for the simultaneous identification of small-molecule activators and inhibitors of HYPE AMPylation. Employing the fluorescence polarization of an ATP analog fluorophore—Fl-ATP—we developed and optimized an efficient, robust assay that monitors HYPE autoAMPylation and is amenable to automated, high-throughput processing of diverse chemical libraries. Challenging our pilot screen with compounds from the LOPAC, Spectrum, MEGx, and NATx libraries yielded 0.3% and 1% hit rates for HYPE activators and inhibitors, respectively. Further, these hits were assessed for dose-dependency and validated via orthogonal biochemical AMPylation assays. We thus present a high-quality HTS assay suitable for tracking HYPE’s enzymatic activity, and the resultant first small-molecule manipulators of HYPE-promoted autoAMPylation.

Haemophilia is one of the most common causes of inherited bleeding disorder resulting from deficiency of coagulation factor VIII or factor IX. Ideally, replacement should be done with factor concentrate. Due to economic constraints associated with its procurement, bleeding episodes are regularly dealt with Fresh Frozen Plasma (FFP) or cryoprecipitate in low-resource countries. This study was carried out to compare the utilization profile and clinical characteristics of haemophilia patients receiving FFP and cryoprecipitate for replacing clotting factor deficiency. This cross-sectional comparative study was conducted in the day care unit of the Department of Transfusion Medicine of Bangabandhu Sheikh Mujib Medical University between 2 groups of haemophilia patients receiving either cryoprecipitate or FFP for treatment. Out of the total 100 haemophilia patients, 50 were treated with cryoprecipitate and 50 with FFP. In FFP group, the majority of patients (48% in cryoprecipitate group and 36% in FFP group) were in the age group of more than 5 to 10 years followed by 11 to 15 years age (24% versus 30%) with a mean SD of age in cryoprecipitate group and FFP group being 11.78±5.61 and 13.42±6.12 years, respectively. About 33 (66.0%) had a history of bleeding following trauma and 32 (64.0%) had a history of spontaneous bleeding among the patients in cryoprecipitate group as a cause of swelling/bleeding and in FFP group, 23 (46.0%) had history of spontaneous bleeding followed by 23 (34.0%) with history of bleeding following trauma. Regarding the type of bleeding, oral bleeding was most common, followed by soft tissue bleeding in both group (40.0% versus 38.0%). Presence of ecchymosis in both groups was statistically significant. The difference in type of haemophilia between the two groups was statistically significant (p<0.001) with a prevalence of haemophilia A of about 88%. The life expectancy of haemophilia patients is increasing dramatically day by day with successful and effective treatment with the appropriate plasma component. Cryoprecipitate is better than FFP as there is less chance of volume overload minimizing leucocyte induced non-haemolytic febrile transfusion reaction and rapid correction of coagulation factor.

Therapeutic plasma exchange (TPE) is an extracorporeal blood purification technique that removes circulating antibodies, toxin and mediators from the plasma of the patients. This study was conducted to assess the safety, type and frequency of complications with regard to the indications and technical aspects of the TPE procedure performed in patients referred with different indications. A total of 127 patients’ clinical data treated with 424 TPE cycles over a period of 6 years from June 2016 to July 2022 at the apheresis unit of the Department of Transfusion Medicine, Bangabandhu Sheikh Mujib Medical University in Bangladesh, were enrolled in this retrospective observational study. The median age was 35.26 years (range 19–63) with a male predominance. The indications were mostly American Society for Apheresis (ASFA) category I (81.895%) and the most common indications were neurological indications including Guillain–Barré Syndrome (GBS) (61.41%), Myasthenia Gravis (MG) (10.24%) followed by Acute Liver Failure (7.09%). Venous access was established with the femoral vein maintaining blood flow at 70 ml/min using acid citrate-dextrose solution A (ACD-A) anticoagulant. TPE was frontline therapy in 55.91% of the patients. Human albumin was used as a replacement fluid in 88% and fresh frozen plasma in 12% of the cycles. Incidence of procedural complication was 49% (n = 206) and majority of them were mild (n = 191; 45%) with only 3% and 0.47% being moderate and severe, respectively.             Mild complications were hypocalcemia (n = 127; 29.95%), anxiety (n = 109; 25.70%), restlessness (n = 68; 16.04%) and hypotension (n = 46; 10.85%) in albumin group and anxiety (n = 31; 7.31%), hypocalcemia (n = 23; 5.42%), muscle cramp (n = 23; 5.42%) in FFP group. Interestingly, complications were almost similar in albumin and FFP groups except for allergic reactions and mild reactions which were mostly manageable and preventable with continuous, cautious monitoring. Our findings suggest that TPE is a relatively safe procedure and severe complications such as hypotension and anaphylaxis is preventable through continuous and dedicated monitoring by expert personnel.