Immunophenotypic Characterization and Ploidy Analysis of Neoplastic Plasma Cells by Multiparametric Flow Cytometry.

Flow cytometric (FCM) immunophenotyping is an important tool for generating diagnostic and prognostic information in plasma cell dyscrasias. This study aimed to evaluate the immunophenotype and ploidy status of plasma cells (PCs) in patients of myeloma and its correlation with other laboratory parameters. Bone marrow of 70 newly diagnosed cases of myeloma were subjected to FCM using a panel of antibodies; CD138, CD38, CD19, CD45, CD28, CD81, CD56, CD200, and CD229. FxCycle Violet (FCV) dye was used for the ploidy analysis of clonal PCs. Median age was 60 years with M:F ratio of 3.2:1. A positive correlation was noted between the morphological and FCM-based PC enumeration (r = 0.4, p = 0.001). Aberrant expression of CD56, CD200, CD28, CD117, CD81 and CD19 and was observed in 88.5%, 77%, 29%, 37%, 23% and 17% cases respectively. Two aberrant antigens were noted in all cases. CD81 + cases had a relatively higher quantity of monoclonal-protein (> 1 g/dl, p < 0.05) and renal insufficiency (Cr > 2 mg/dl, p < 0.05) as compared to the CD81- cases. CD229 was expressed in all the cases, with a median MFI in PCs significantly higher than other hematopoietic elements. Hyperdiploid PCs (median DI-1.59, range, 1.16-2.6) were noted in 80% cases (n = 48), diploid/ near-hyperdiploid PCs in 8% (n = 5) cases and hypodiploidy in 3% (n = 1) cases. Bright CD56/CD200 and CD45- can identify abnormal PC in the majority of the cases. CD81 appears to correlate with disease burden and might be useful as a prognostic marker. CD229 is a reliable gating marker for plasma cells. Ploidy analysis may be incorporated in routine workup to guide in the identification of patients with poor prognosis. Supplementary Information: The online version contains supplementary material available at 10.1007/s12288-021-01477-y.

Financial support to the poor for the detection of smear-negative pulmonary and extra-pulmonary TB in Bangladesh.

BACKGROUND: The study was conducted in BRAC-administered areas of the Bangladesh National Tuberculosis (TB) Programme (42 of 64 districts). According to the 2013-2017 financial support scheme, direct costs due to TB diagnosis were reimbursed among economically disadvantaged people with presumptive smear-negative pulmonary (PTB) and extrapulmonary TB (EPTB).OBJECTIVE: To describe the implementation of the scheme and associated changes in case notification.DESIGN: This was a descriptive study involving programme data.RESULTS: Between 2013 and 2017, persons reimbursed reduced from 125 680 to 88 763, and the case detection ratio increased from 18% to 24%. The number of patients with presumptive EPTB who were reimbursed decreased from 5024 to 3484. More than 95% were reimbursed for chest radiograph, fine-needle aspiration cytology and biopsy. However, large numbers of ancillary investigations were also reimbursed. During 2013-2017, the observed national quarterly new smear-negative PTB case notification rates (CNRs) were significantly higher than the forecasted CNRs (based on CNR trends during 2008-2012). New EPTB and all form TB CNRs increased but not significantly.CONCLUSION: Implementation of the financial support scheme was accompanied by a significant improvement in new, smear-negative PTB notification. The absence of a comparison arm was a key limitation, but comparison was not possible as the scheme was implemented in all districts.

The triple-negative (CD34-/HLA-DR-/CD11b-) profile rapidly and specifically identifies an acute promyelocytic leukemia.

INTRODUCTION: The genetic testing to confirm or rule out an acute promyelocytic leukemia (APL) typically takes a minimum of 24-72 hours. Flow cytometric immunophenotyping (FCI) on the other hand provides rapid and objective information to differentiate APL from non-APL. METHODS: FCI features, with single-tube 8-color combination using CD45, CD34, HAL-DR, CD11b, CD13, CD33, and CD117 and CD64, were compared for the 30 consecutive APL and 30 non-APL acute myeloid leukemia (AML) cases which morphologically mimicked an APL. The diagnosis was confirmed by cytogenetic or molecular genetic testing in the form of t (15:17) (q22; q21)/variant translocations or PML-RARA fusion transcript analysis. RESULTS: The APL cells lacked CD34, HLA-DR, and CD11b in 90%, 90%, and 93.3% cases, respectively. Myeloid antigens such as CD33, CD13, CD117, and CD64 were expressed in 96.7%, 96.7%, 76.7%, and 70% cases, respectively. The dual negative profiles, CD34-/HLA-DR- or HLA-DR-/CD11b-, were noted in 90% and 93.3% cases. The triple-negative (CD34-/HLA-DR-/CD11b-) profile was noted in 90% of the cases. The sensitivity, specificity, and positive predictive value (PPV) of CD34-/HLA-DR- and HLA-DR-/CD11b- profiles for the diagnosis of APL were found to be 90%, 80% & 81.1% and 93.3%, 86.7%& 87.5%, respectively. Combining the above two profiles resulted in a triple-negative profile (CD34-, HLA-DR- and CD11b-), which had a better specificity (93.3%) and positive predictive value (93.1%), with similar sensitivity. CONCLUSION: FCI is a rapid and reliable modality for the diagnosis of an APL. The triple-negative profile (CD34-/HLA-DR-/CD11b-) rapidly and specifically identifies an APL case.

Inhibition of carrageenin-induced pedal oedema in rats by immobilisation stress.

The effect of immobilisation stress on acute pedal inflammation induced by carrageenin, and the mechanism of stress-induced anti-inflammatory effect, were investigated in male Wistar strain albino rats. Carrageenin-induced pedal inflammation oedema was attenuated by immobilisation stress in a time-dependent manner, when the rats were restrained for 30 min, 1 h, and 2 h immediately after the induction of the inflammation. Pentobarbitone exhibited significant anti-inflammatory effect of its own in an anaesthetic dose and also inhibited stress (1 h)-induced attenuation of the inflammation. Likewise, lignocaine, injected behind the knee joint of the inflamed limb, attenuated the inflammation and also inhibited the stress-induced anti-inflammatory effect. These findings indicate the importance of the central nervous system (CNS) and the afferent/efferent neural pathways from and to the inflammatory site, in inflammation and in stress-induced anti-inflammatory effect. Earlier studies from this laboratory have shown that the central noradrenergic, histaminergic, serotonergic and GABA-ergic neurotransmitter systems have a modulatory anti-inflammatory effect on carrageenin-induced pedal oedema. Since all these neurotransmitter systems have been reported to be activated by stress, their role was assessed in the inflammation-attenuation effect of immobilisation stress. The present studies indicate that, of these neurotransmitters, only the central noradrenergic system is involved in the anti-oedema effect of stress. Endogenous opioid peptides may also be involved in the stress-inflammation interaction, since naloxone inhibited the stress effect. Bilateral adrenalectomy and peripheral chemical sympathectomy, induced by i.p. administration of 6-hydroxydopamine, augmented carrageenin oedema and antagonised the stress-induced anti-inflammatory effect. However, metyrapone, an inhibitor of endogenous corticoid synthesis, failed to inhibit the stress effect. These findings indicate that the sympatho-medullary system, which is known to be activated during stress, is responsible for the observed anti-inflammatory effect of immobilisation stress, rather than augmented release of adrenal corticoids. It is suggested that the observed inflammation reducing effect of immobilisation stress is a consequence of increased central noradrenergic and peripheral sympatho-medullary activity.

Effect of some centrally administered putative amino acid neurotransmitters on carrageenan-induced paw oedema in rats.

The possible modulatory effect of central amino acid neurotransmitters on carrageenan-induced paw inflammation has been investigated in rats. Eight putative amino acid neurotransmitters were administered intracerebroventricularly and their effect on the peripheral inflammation was noted. The inhibitory amino acid transmitters, GABA, glycine and taurine attenuated the peripheral oedema, while the excitatory amino acid transmitters, glutamic acid and aspartic acid had a pro-inflammatory effect. However, the other putative amino acid neurotransmitters, proline and alanine (inhibitory) and cysteic acid (excitatory) did not affect carrageenan-induced oedema.

Effect of lycoriside, an acylglucosyloxy alkaloid, on mast cells.

The effect of lycoriside, an acylglucosyloxy alkaloid from Crinum asiaticum Linn, (family Amaryllidaceae), with or without sitosterol-3-O-ß-D-glucoside, was studied on the rate of degranulation of peritoneal mast cells of albino rats. Lycoriside, at lower concentrations (1-20 µg/ml), in vitro, produced statistically significant protection against Tween 80-induced degranulation, as also to sensitized mast cells challenged with an antigen (horse serum). It also provided protection against compound 48/80-induced degranulation of mast cells when administered in vivo (1-5 mg/kg, po). At higher concentrations (100 µg/ml and above), in vitro, however, it had a mast-cell degranulation effect per se. The addition of sitosterol-3-O-ß-D-glucoside to lycoriside did not modify the effect of the latter compound. The mechanism of the dual response elicited by lycoriside is appraised in view of a concentration-dependent anti- or prerelease effect on mast-cell mediators.