Collage of cases and brief review of the laboratory diagnosis and molecular testing in autoimmune haemolytic anaemia.

Autoimmune haemolytic anaemia (AIHA) is an acquired heterogenous clinical entity with variable presentations like acute haemolysis or mild, chronic haemolysis compounded with acute exacerbation in winters or fatal uncompensated haemolysis. A step-wise approach to the diagnosis and characterisation of AIHA should be undertaken, firstly the diagnosis of haemolysis followed by the establishment of immune nature with the aid of direct agglutination tests (DAT). Simultaneously the other causes of immune haemolysis need to be excluded too. In light of advancements in diagnostics, a wide array of investigations can be used like absolute reticulocyte count, bone marrow responsiveness index to establish the evidence of haemolysis, sensitive gel technology, enhanced DAT assays, e.g., modified DAT with low ionic strength saline solution (LISS) at 4°C, DAT assays utilizing reagents such as anti-IgA and anti-IgM and DAT by flowcytometry, to detect RBC bound autoantibodies (Abs) and monospecific DAT to establish immune causes of haemolysis and characterisation of the autoantibodies. The compensatory role of bone marrow and synchronous pathologies like clonal lymphoproliferation, dyserythropoiesis, fibrosis are important factors in the evolution of the disease and aid in the customisation of treatment modalities. The laboratory work up should aim to diagnose underlying diseases like chronic lymphoproliferative disorders, autoimmune disorders and infectious diseases. Also, tests like autoimmune lymphoproliferative syndromes (ALPS) screening panel and Next-generation sequencing (NGS) panel for RBC membrane disorders, RBC enzymopathies, and congenital dyserythropoietic aneamia have found their place. It is incumbent upon the clinicians to use the all-available diagnostic modalities for the accurate diagnosis, prognostication and customisation of the therapy.

Assessment of pre and post donation changes in hematological parameters and serum calcium and magnesium levels in plateletpheresis donors.

Background: Single donor platelet products are preferred over random donor platelet products due to several advantages. However, safety issues with regard to post procedure hematological decrements and serum calcium and magnesium levels in donors undergoing plateletpheresis have been explored minimally. Aims: This prospective study was done to analyze the effects of plateletpheresis on donor's hematological parameters and serum calcium and magnesium levels. Settings and Design: It is a prospective, cross-sectional study conducted in the department of immunohematology and blood transfusion. Material and Methods: This study was undertaken on 150 healthy plateletpheresis donors over a period of 1 year. Blood samples were collected from each donor before and after the procedure, one in ethylenediaminetetraacetic acid (EDTA) vial for estimation of hematological parameters and another in plain vial for serum calcium and magnesium levels. Statistical Analysis Used: Paired t-test was used to analyze the data. Results: This study included donors in the age group of 18 to 60 years who underwent plateletpheresis on Haemonetics model of a machine (MCS) + intermittent flow cell separator. A statistically significant increase was observed in mean post procedural Hb (0.95%), Hct (0.7%), and red blood cell (RBC) count (1.3%). There was a decrease in mean post procedural platelet count (27.5%), white blood cell (WBC) count (4.02%), mean serum calcium (1.5%), and serum magnesium (5.1%), which was statistically highly significant (P < 0.001). No significant change was observed in post procedural mean platelet volume (MPV) and platelet distribution width (PDW). Conclusion: Amid the need of increased demand for plateletpheresis, donor safety must be ensured. Failing to do so can be detrimental to blood supply chain, hence stringent programs for post donation screening of plateletpheresis donors need to be established.

Acquired-B phenomenon in a neonate presenting with necrotizing enterocolitis.

OBJECTIVE: We are reporting a case of acquired B phenomenon in a 14 days old pre-term baby presenting with necrotizing enterocolitis (NEC) where it was detected as an ABO discrepancy. METHOD: The forward and reverse grouping was done using tube technique as well as column agglutination technique (ABD card and LISS Coombs AHG gel cards, Biorad, Switzerland). The direct antiglobulin test (DAT) was also done on gel card. Further work up was done using acidified (0.1 N HCl) anti-B (pH 6-6.5) polyclonal anti-B (from group A donors) for resolution of ABO discrepancy. RESULTS: The forward grouping was AB RhD positive with a 'mixed-field' agglutination with anti-B. The reverse grouping using pooled A cells, B cells and O cells gave 'negative' result with both the techniques. Anti-A1 gave 2+ agglutination, anti-H gave 1+ agglutination and autologous control was negative. The DAT was also negative. The patient's previous group on day 1 of life was A RhD positive and had received 1 PRBC (pedibag) transfusion. As the patient's blood culture was positive for Klebsiella pneumoniae, it could have contributed to 'acquired-B' phenomenon. Acidified (0.1 N HCl) anti-B (pH 6-6.5) and polyclonal anti-B yielded no agglutination with patient red cells. The patient was kept on antibiotics subsequently and the blood group after 3 weeks was found to be A RhD positive. CONCLUSION: This case of 'acquired-B' phenomenon in a neonate with NEC emphasizes the relevance of clinical findings as a guide to resolve blood group discrepancy and deciding further transfusion strategy.

Regenerative efficacy of therapeutic quality platelet-rich plasma injections versus phonophoresis with kinesiotaping for the treatment of chronic plantar fasciitis: A prospective randomized pilot study.

BACKGROUND: Plantar fasciitis (PF) a common chronic musculoskeletal pain routinely diagnosed and treated in rehabilitation practices. When conservative management fails in this degenerative disease, local injections of corticosteroids, platelet rich plasma (PRP), botulinum toxin, extracorporeal shockwave therapy, surgical release are used. In our prospective randomized pilot study we compared the regenerative efficacy of Platelet Rich Plasma vs Kinesiotaping with phonophoresis who were resistant to conservative management of PF. MATERIALS AND METHODS: Sixty-four chronic plantar fasciitis patients nonresponding to conservative management were evaluated for two interventions.36 patients received ultrasound guided 2.5 ml autologous platelet rich plasma (PRP) injection and 28 participant received phonophoresis and total10 Kinesiotaping on alternate days. 54 participants 33 in PRP intervention group and 21 in KT group were analyzed, by Numerical Rating Scale (NRS), plantar fascia thickness (ultrasound guided) and disability and activity limitation measured by foot function index in every two weeks up to 6 months. RESULTS: Post intervention assessment at 2 weeks revealed improvement in pain relief was better in Kinesiotaping group (NRS-4.619) as compared to PRP group (NRS- 6.061). But evaluation at 12 and 24 weeks showed statistically significant improvement in NRS and Foot function index in PRP group than in Kinesiotaping. Similarly, at the end of 24-week improvement in foot function index (FFI) was more in PRP group (P<0.0001). At end of 12 and 24 weeks there was significant reduction in plantar fascia thickness in PRP group (p<0.0001) as compared to KT group (p<0.05). CONCLUSION: Our study concluded that therapeutic quality autologous PRP injection (1x106 platelets/µl) has regenerative effect with long and better efficacy in pain management of chronic recalcitrant plantar fasciitis than Phonophoresis and Kinesiotaping.

Daily plasma exchange in severe Guillain-Barré syndrome helps in early weaning from ventilator: A lesson from a case.

Intravenous immunoglobulin (IVIG) and therapeutic plasma exchange (TPE) are well-known therapeutic modalities in Guillain-Barré syndrome (GBS). In developing countries like India, where plasma-derived products (IVIG) are not easily available, and affordable TPE is preferred. Here, we reported a case of severe GBS, who was treated with daily plasma exchange (PLEX) rather than recommended alternate day schedule. A 16-year-old male adolescent of severe GBS, i.e., on mechanical ventilator was treated with the plasmapheresis regimen consisted of removal of 1.3 plasma volumes in each cycle for total of five cycles, on daily basis. The patient's condition started improving after three cycles of TPE with power in the upper limbs 4/5 and lower limbs 3/5 and completely weaned off from ventilator after the 4th TPE, i.e. the 4th day of admission. This case emphasizes the need of daily PLEX regimen particularly in severe GBS patients because early weaning from ventilator reduces the ventilator-associated complications, hospital stay as wells as less morbidity, and mortality in severe GBS.