Morphological Changes in Blood Cells as Indicators for Disease Progression in COVID 19

A novel highly pathogenic human corona virus (COVID19) has been recently recognised in Wuhan, China as the cause of corona disease outbreak. It has rapidly spread from China to various countries across the world evolving as a pandemic. In our study we have categorized the covid positive patients into mild, moderate and severe based on the clinical criteria suggested by WHO. The coagulation parameters of the patients were analysed and documented. A peripheral smear was made for every patient and the morphological changes in blood cells were documented. The peripheral smear findings were then correlated with the disease stage and coagulation parameters. There were significant differences in the total WBC count and the differential WBC count between stages 1 &2 and stages 1 & 3 (p<0.005). Leucocytosis, neutrophilia and toxic changes in neutrophils were seen in severe stage of the disease and in covid coagulopathy suggesting these are important indicators of disease severity. Schistocytes an important finding in any other coagulopathy was not present in covid associated coagulopathy. Activated lymphocytes was found to be the most common morphological presentation seen in all covid patients irrespective of the disease stage whereas plasmacytoid lymphocytes was an important finding in severe stage disease. Monocyte cytoplasmic vacuoles, large/giant platelets were other morphological findings observed but these findings did not have any significant correlation with disease stage. Since follow up smears of the same patient were not analysed during disease progression and also post recovery, additional research in this field will provide further insights.

Partial HELLP syndrome: case report

HELLP syndrome is a complication in pregnancy clustered by haemolysis, elevated liver enzymes, and a low platelet count. It is seen as a serious complication of preeclampsia and eclampsia. Serious manifestations like haemorrhage, infarction, rupture and other hepatic manifestations are usually associated with it. In this case study, 29 years old primigravida is a booked case admitted in ward at 39 weeks 1 day with decreased fetal movement for 2 days. No history of pain abdomen, bleeding per vaginum, discharge per vaginum. Her blood pressure records at the time of admission was 110/72 mmHg and she was normotensive throughout pregnancy. Urine routine examination was negative for urinary protein. However, blood tests showed platelet count of 66,1000/cumm, with ALT of 174 U/L and AST of 123 U/L on peripheral blood film. RBC were predominantly normocytic, normochromic with few macrocytes. WBC has normal morphology. Platelets were reduced on smear. Giant platelets were seen. Ursodeoxycholic acid 300 mg 12 hourly were given to the patient and 3 doses of vitamin K I/M 24 hourly. She was delivered by cesarean section which was performed due to failure of progression of labor with a deflexed head. There was presence of retroplacental clot of 4×3 cm indicating placental abruption, a complication of HELLP syndrome. From this we conclude that we should be careful in suspecting complications of full blown diseases even when the patients are asymptomatic but have atypical laboratory findings.

Improving Residents' Knowledge of Peripheral Smears

Background: A peripheral smear is an important diagnostic tool, inexpensive, reliable and quick. It reflects the functional status of the bone marrow and can identify many blood disorders, some of which are life threatening and very time sensitive. Peripheral smears are useful to assess cytopenic states (eg, anemia, leukopenia, thrombocytopenia) or identify hematologic emergencies such as thrombocytopenic purpura/hemolytic uremic syndrome, acute myeloid leukemia, and disseminated intravascular coagulation etc. Sometimes it is sufficient to make a diagnosis solely based on them which can be life saving when time is of the essence. Based on our literature search, there are no studies to assess internal medicine residents' knowledge and understanding of peripheral smears. Methods: We created a multiple-choice questions survey of 13 questions for residents to fill out, involving common findings on peripheral smears and those found in hematologic emergencies that every resident should be aware of. Response rate in all three program years (PGY) was 100%. 16/16 PGY-1, 10/10 PGY-2 and 9/9 PGY-3 completed the survey. Results: Average pre-intervention score for PGY-1 was 42%, PGY-2 was 34% and PGY-3 was 30%. Questions most commonly wrong were regarding target cells, metamyelocytes, giant platelets and schistocytes. Most were correct about microcytosis and Auer rods. We then conducted the same survey weeks after our intervention, which was interactive, small group didactics sessions on peripheral smears. We compared results of the survey before and after didactics to check for improvement. Post-intervention score for PGY-1 was 98%, PGY-2 and PGY-3 were 96%. Conclusions: Most medical residents are not aware of common interpretations of peripheral smears and this does not seem to improve with each progressive year. Improving residents' knowledge of peripheral smears is a cost effective and quick measure that can improve patient care, in addition to potentially improving internal medicine board results. In the future, we aim to make a curriculum for peripheral smear review for residents. We can compare results of the survey with hematology oncology fellows. Disclosures No relevant conflicts of interest to declare.

Pure Smooth Endoplasmic Reticulum (SER) Inclusions in Granulocytes of MYH9-Related Disorder Patients

MYH9-related disorders are a group of disorders (May-Hegglin anomaly, Fechtner syndrome, Sebastian syndrome) characterized by macrothrombocytopenia and ribosome inclusions in granulocytes. They are caused by mutations in the MYH9 gene and, depending on the location of the mutation, are associated with extrahematological pathologies (sensorineural hearing loss, progressive nephropathy, presenile cataracts) of variable intensity. The inclusions are formed by single and clustered ribosomes and are partially surrounded by segments of rough endoplasmic reticulum (RER). Typical inclusions in May-Hegglin anomaly are spindle-shaped, well-defined, and contain longitudinal filaments. Those found in Sebastian and Fechter syndromes are round or oval, have no filaments and, sometimes, show cross-striated arrangement of the ribosome aggregates. When reviewing the electron micrographs of the granulocyte inclusions of 10 patients with MYH9-related disorders previously published (Pujol-Moix et al. Haematologica 2004;89:330-337) we observed a special type of inclusion not described in the article. This inclusion has the typical appearance of smooth endoplasmic reticulum (SER) clusters, that is, a meshwork of branching tubules of SER without limiting membranes and no other structures such as ribosomes or RER. In 3 of the patients reviewed, in addition to the typical inclusions of MYH9-related disorder, some granulocytes with SER clusters were observed. These clusters were rounded or oval in shape and, when present, were 1 or 2 per cell (Fig. 1). The patients who presented granulocyte SER clusters are those identified as B4, C6 and D8 in the Haematologica article and are briefly described in Table 1. The patients presenting with SER clusters showed different characteristics such as age, bleeding, platelet counts, and proportion of giant platelets. Therefore, we cannot relate the presence of SER clusters to any clinical or biological characteristic of patients. It should be noticed that we did not find SER clusters in any patient with May-Hegglin anomaly. However, the small number of cases studied does not allow any conclusions to be drawn. SER clusters have been described in different types of cells such as hepatocytes or kidney tubular epitelial cells (Ghadially FN. Ultrastructural Pathology of the Cell and Matrix. Butterwords. London, 1988, pp 422-427). Normal granulocytes (neutrophils, eosinophils, and basophils) besides their own specific granules, also contain other general subcellular structures, including dispersed SER membranes. However, SER membranes forming clusters in these cells have not been previously described. In addition, SER clusters do not seem to correspond to evolved forms of typical MYH9 inclusions, for example by degranulation of the RER, based on the observations that typical inclusions show abundant ribosomes but few RER segments, no appreciable ribosomes are observed in the SER clusters, and the morphology of the tubules forming the SER and RER is different. We suggest that SER clusters could be considered a new type of inclusion in MYH9-related disorders. Figure 1 Figure 1. Disclosures No relevant conflicts of interest to declare.

Red Blood Cell Morphological Changes and Enlarged Platelets Found in Idiopathic Multicentric Castleman Disease

BACKGROUND Idiopathic multicentric Castleman disease (iMCD) is a rare polyclonal lymphoproliferative disorder characterized by systemic inflammatory symptoms, cytopenias, generalized lymphadenopathy, and multiple organ system dysfunction. iMCD is subclassified into iMCD-TAFRO, with thrombocytopenia, anasarca, fever/elevated C-reactive protein, reticulin myelofibrosis/renal failure, and organomegaly, and iMCD-NOS (not otherwise specified), with high platelet counts and hypergammaglobulinemia. Despite recent diagnostic guidelines, iMCD diagnosis remains challenging because many of the histopathologic features and clinical/laboratory abnormalities are non-specific. Characterization of easily measurable findings from minimally invasive procedures, such as peripheral blood smear, could help to improve the accuracy of diagnosis as well as potentially providing insights into disease pathogenesis. Peripheral blood smears are often performed during routine clinical evaluation and enable morphologic evaluation of red blood cells (RBCs) and white blood cells (WBCs). In this study, we systematically characterized peripheral blood smear morphologic findings in iMCD for the first time. METHODS Peripheral blood smear morphologic findings in medical records from 68 iMCD patients, who were enrolled in an international registry for CD patients and had been confirmed to meet iMCD criteria by an expert panel, were reviewed and analyzed. Patients were also categorized into iMCD-TAFRO (N=39) or iMCD-NOS (N=29) based on clinical and laboratory data. Two-sample proportion tests were used to compare the frequency of features between iMCD-TAFRO and iMCD-NOS, when the abnormality was present in ≥ 5 subjects in both subtypes. P-value correction was not performed as findings are preliminary and hypothesis generating. RESULTS Among the 68 iMCD patients, average age was 35.8 years (14-61); 46% were female, 54% were male; racial distribution was 62% white; 10% Black; 15% Asian; 13% other. The most common peripheral blood smear findings across all iMCD patients included abnormalities in RBC size and color, with anisocytosis (58.8%), polychromasia (57.4%), hypochromia (55.9%), and microcytosis (50.0%) being the most prevalent findings in the overall cohort (Table 1). Other RBC abnormalities were observed but less frequently, including abnormalities in shape such as poikilocytosis (44.1%). Giant or large platelets were found in 47% of iMCD cases. Abnormalities in WBCs, including toxic granulation, atypical lymphocytes, and toxic vacuolization, were detected less frequently. When comparing the two clinical subtypes, patients with iMCD-TAFRO had more frequent anisocytosis (p=0.043), hypochromia (p=0.002), polychromasia (p=0.022), and giant/large platelets (p&lt;0.001) versus iMCD-NOS. DISCUSSION Here, we have characterized peripheral blood morphologic abnormalities in iMCD for the first time. The most common findings in this cohort included abnormalities in RBC size and color. These abnormalities can be found in a number of conditions, including myelofibrosis, autoimmune diseases, and other inflammatory conditions. In iMCD, they are likely related to cytokine-driven anemia of chronic inflammation. The presence of large or giant platelets in nearly half of iMCD patients, with significantly more in iMCD-TAFRO than iMCD-NOS, is particularly notable since this has not been described in iMCD. These enlarged platelets, which can be found in immune thrombocytopenic purpura, myeloproliferative disorders, pseudothrombocytopenia, and inherited platelet disorders, may reflect hyperactivity of megakaryocytes and/or early release from the bone marrow in response to peripheral platelet sequestration. Though less common in this cohort, there are additional changes in RBC shape and WBC features that may be more specific to iMCD and potentially informative in increasing the index of suspicion for iMCD. These findings suggest that the peripheral blood smear may be able to support the diagnosis of iMCD and result in faster treatment administration. Future work is needed to determine whether the constellation of findings in the peripheral blood identified herein is unique to iMCD or seen in various other infectious, malignant, and autoimmune diseases. Figure 1 Figure 1. Disclosures Fajgenbaum: Pfizer: Other: Study drug for clinical trial of sirolimus; EUSA Pharma: Research Funding; N/A: Other: Holds pending provisional patents for 'Methods of treating idiopathic multicentric Castleman disease with JAK1/2 inhibition' and 'Discovery and validation of a novel subgroup and therapeutic target in idiopathic multicentric Castleman disease'.

Platelet Phagocytosis by Neutrophils, Platelets Lack of Granules and Giant Platelets Result in Pseudothrombocytopenia

Pseudothrombocytopenia (PTCP) is a condition in which the decreased platelet count does not agree with the clinical status of the patient, can lead to misdiagnose, unnecessary tests, unnecessary treatment. The present case describes a 73-year-old man suffered with pulmonary tuberculosis, treated with anti-tuberculosis therapy (isoniazid, rifampicin, pyrazinamide, ethambutol, 2HRZE/4HR). One month later, the patient had a significant decrease in platelets (101 to 56 x109/L). Peripheral blood smear showed that 28% platelets were phagocytosis by neutrophils, 26% platelets were lack of granules and 6% platelets’ volume increased significantly. When the anticoagulant was changed from EDTA to sodium citrate, there was no change in the above phenomenon. By manual count, the value of platelets was 113 x 109/L. After the completion of anti-tuberculosis therapy, platelet morphology gradually returned to normal. HRZE treatment may cause platelet morphology abnormal, resulting in PTCP. In such cases, we should regularly review the peripheral blood smear to ensure the accuracy of the results and avoid unnecessary examination and treatment. The emergence of PTCP may does not mean the presence of specific disorders.

Evaluation of platelet parameters in cerebral stroke in patients with HIV infection

The article reflects the results of studying platelet parameters in HIV-positive patients with different types of stroke.Aim. To identify changes in laboratory parameters of a complete blood count which characterize the morphofunctional features of platelets in stroke among HIV-positive patients.Materials and methods. 110 HIV-positive patients who received treatment for stroke in hospitals of the Tyumen region were examined. The study of blood parameters was carried out at the analyzer Sysmex XE2100 (Japan). Blood sampling was carried out on the day of patients admission.The number of platelets and platelet indices were analyzed: MPV – mean platelet volume, PDW – platelet distribution width, PCT – plateletcrit and P-LCR – platelet large cell ratio. The control group consisted of 117 patients. The signifcance of the differences was determined at the twotailed signifcance level of p < 0.05.Results. There was a signifcant decrease in the number of platelets (p < 0.05), in average, on 34.3% among patients with hemorrhages and HIV infection. Among patients with ischemic stroke this decrease was less pronounced (p = 0.05). A signifcant decrease in plateletcrit was established among patients with intracranial hemorrhages, while it did not change signifcantly among patients with cerebral infarction. During evaluation of other platelet parameters, no signifcant differences were found between patients in experimental and control groups. The coeffcient of giant platelets prevailed by one and a half times in patients with hemorrhages associated with HIV infection, which turned out to be beyond the statistical signifcance.Conclusion. The presence of HIV infection leads to a more pronounced, reliable decrease in the number of platelets and plateletcrit among patients that have acute phase of the development of hemorrhagic stroke than in patients with cerebral infarction. The development of intracranial hemorrhage among HIV-positive patients is characterized by an increase of blood platelets with a high volume, the level of which increased by one and a half times being beyond the statistical signifcance and having as a leading mechanism the intensifcation of platelet formation in the bone marrow.

A Mathematical Model of Platelet Antiparasitic Action in Erythrocytes and Merozoites During Malaria Infection

Platelets have been seen traditionally as fragments of blood mediating coagulation. However, evidence during malaria infection suggests that platelets also act against merozoites, an infectious form of malaria in the bloodstream, and megakaryocytes can release giant platelets with a larger volume than normal platelets. We propose a mathematical model to study the interaction between red blood cells, merozoites, and platelets during malaria infection. We analyzed two cases of the interaction of platelets with malaria infection. In the first one, we considered the isolated action of normal platelets and, in the second one, the joint antiparasitic action of both normal and giant platelets. Numerical simulations were performed to evaluate the stability of the equilibrium points of the system of equations. The model showed that the isolated antiparasitic action of normal platelets corroborates malaria infection control. However, the system can converge to a presence-merozoite equilibrium point, or an oscillatory behavior may appear. The joint antiparasitic action of both normal and giant platelets eliminated the oscillatory behavior and drove the dynamics to converge to lower parasitic concentration than the case of isolated action of normal platelets. Moreover, the joint antiparasitic action of platelets proved more easily capable of eliminating the infection.

The curious incident of a cavum velum interpositum cyst in twins of a mother carrying May-Hegglin anomaly: a case report and short literature review

Background May-Hegglin anomaly is an autosomal dominant inherited condition, characterized by thrombocytopenia, giant platelets and Dohle-like bodies. Incidence is unknown and affected individuals can show from mild to moderate-severe haemorrhagic symptoms. The cyst of cavum veli interpositi (a virtual space filled with fluid within the third ventricle) is rarely reported in the foetal period. Furthermore, it is unclear whether isolated cavum veli interpositi cysts are a normal variant or developmental malformations. The simultaneous presence of these two anomalies was never described. Case presentation We describe a very rare case of a twin monochorionic pregnancy in a woman with the May-Hegglin anomaly, whose foetuses carried cavum veli interpositi cysts. Since childhood, our patient had shown macro-thrombocytopenia, deafness and bleeding (epistaxis and menorrhagia), but she was misdiagnosed until the age of 30 years when our Centre identified a de novo allelic variant in the gene MYH9 coding for the non-muscle myosin heavy chain IIa. Our patient bled neither during the pregnancy, nor in the peripartum period. Children are now eight-months-old and have never bled, although both inherited the MYH9 variant and have thrombocytopenia with giant platelets. Furthermore, none of them developed psychomotor disorders. Conclusions To the best of our knowledge, this is the sixth case of twin pregnancy in a woman carrying May-Hegglin anomaly and the first one with cavum veli interpositi cysts in the neonates. We speculate that MYH9 could have, at least in part, played a role in the development of both conditions, as this gene has a pleiotropic effect.