A comparative study between light transmission aggregometry and flow cytometric platelet aggregation test for the identification of platelet function defects in patients with bleeding

Background@#Platelet aggregation studies using conventional light transmission aggregometry (LTA) have several disadvantages and require strict pre-analytical measures for reliable results.We aimed to examine the utility of flow cytometric platelet aggregation (FCA) assay in detecting platelet function defects (PFDs) in patients with a history of bleeding symptoms. @*Methods@#Sixty-four participants (24 patients and 40 healthy controls) were included in this study.LTA and FCA assay were performed simultaneously in patients and healthy controls. In the FCA assay, two portions of platelets from the same individual were labeled separately with CD31-FITC and CD31-PE. After mixing and stimulation with agonists, the double-colored platelet aggregates were visualized using a flow cytometer. The results generated using the two techniques were compared and correlated. @*Results@#The patients’ median age was 17 years (range, 3‒72 yr) with a male-to-female ratio of 1:1.7. There was substantial agreement between LTA and FCA assay in detecting a PFD (κ=0.792). Four patients showing a Glanzmann thrombasthenia-like pattern on LTA exhibited an abnormal FCA. A functional defect in collagen binding was detected on the FCA assay conducted in two immune thrombocytopenic patients with severe bleeding. @*Conclusion@#FCA assay can be used to identify functional defects in platelets, with potential applications in thrombocytopenic individuals. It also facilitates the diagnosis of inherited bleeding disorders with platelet defects.

A comparative study between light transmission aggregometry and flow cytometric platelet aggregation test for the identification of platelet function defects in patients with bleeding

Background@#Platelet aggregation studies using conventional light transmission aggregometry (LTA) have several disadvantages and require strict pre-analytical measures for reliable results.We aimed to examine the utility of flow cytometric platelet aggregation (FCA) assay in detecting platelet function defects (PFDs) in patients with a history of bleeding symptoms. @*Methods@#Sixty-four participants (24 patients and 40 healthy controls) were included in this study.LTA and FCA assay were performed simultaneously in patients and healthy controls. In the FCA assay, two portions of platelets from the same individual were labeled separately with CD31-FITC and CD31-PE. After mixing and stimulation with agonists, the double-colored platelet aggregates were visualized using a flow cytometer. The results generated using the two techniques were compared and correlated. @*Results@#The patients’ median age was 17 years (range, 3‒72 yr) with a male-to-female ratio of 1:1.7. There was substantial agreement between LTA and FCA assay in detecting a PFD (κ=0.792). Four patients showing a Glanzmann thrombasthenia-like pattern on LTA exhibited an abnormal FCA. A functional defect in collagen binding was detected on the FCA assay conducted in two immune thrombocytopenic patients with severe bleeding. @*Conclusion@#FCA assay can be used to identify functional defects in platelets, with potential applications in thrombocytopenic individuals. It also facilitates the diagnosis of inherited bleeding disorders with platelet defects.

Role of Genetic Variants and Gene Expression in the Susceptibility and Severity of COVID-19

Since its first report in December 2019, coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has rapidly emerged as a pandemic affecting nearly all countries worldwide. As the COVID-19 pandemic progresses, the need to identify genetic risk factors for susceptibility to this serious illness has emerged. Host genetic factors, along with other risk factors may help determine susceptibility to respiratory tract infections. It is hypothesized that the ACE2 gene, encoding angiotensin-converting enzyme 2 (ACE2), is a genetic risk factor for SARS-CoV-2 infection and is required by the virus to enter cells. Together with ACE2, transmembrane protease serine 2 (TMPRSS2) and dipeptidyl peptidase-4 (DPP4) also play an important role in disease severity. Evaluating the role of genetic variants in determining the direction of respiratory infections will help identify potential drug target candidates for further study in COVID-19 patients. We have summarized the latest reports demonstrating that ACE2 variants, their expression, and epigenetic factors may influence an individual’s susceptibility to SARSCoV-2 infection and disease outcome.

microRNAs in Mycobacterial Infection: Modulation of Host Immune Response and Apoptotic Pathways

Our current knowledge of mycobacterial infections in humans has progressively increased over the past few decades. The infection of Mycobacterium tuberculosis causes tuberculosis (TB) disease, which has reasoned for excessive morbidity and mortality worldwide, and has become a foremost issue of health problem globally. Mycobacterium leprae, another member of the family Mycobacteriaceae, is responsible for causing a chronic disease known as leprosy that mainly affects mucosa of the upper respiratory tract, skin, peripheral nerves, and eyes. Ample amount of existing data suggests that pathogenic mycobacteria have skilled in utilizing different mechanisms to escape or offset the host immune responses. They hijack the machinery of immune cells through the modulation of microRNAs (miRs), which regulate gene expression and immune responses of the host. Evidence shows that miRs have now gained considerable attention in the research, owing to their involvement in a broad range of inflammatory processes that are further implicated in the pathogenesis of several diseases. However, the knowledge of functions of miRs during mycobacterial infections remains limited. This review summarises recent findings of differential expression of miRs, which are used to good advantage by mycobacteria in offsetting host immune responses generated against them.

Multifocal large aggregates of pseudo-Gaucher cells in chronic myeloid leukemia

No abstract available.

Role of liver transplantation in severe alcoholic hepatitis

Severe alcoholic hepatitis has very high short term mortality and corticosteroids have been the mainstay of treatment for decades. Patients with Lille score >0.45 are considered non-responders to steroids and have poor outcome. Recently Orthotopic Liver Transplantation (OLT) is being increasingly used as rescue treatment for these patients, without waiting for 6 months of abstinence. Liver transplant is the only rescue treatment which can potentially provide long term benefit for patients who are steroid non-responders. However, with scarcity of organs being a concern, all patients of severe alcoholic hepatitis cannot be chosen for transplantation in an arbitrary way. There is a need for development of predictive tools and objective protocols to select patients who can justify the use of precious liver grafts. With a stringent criteria for selection of patients receiving the graft, liver transplantation in severe alcoholic hepatitis can become a viable rescue therapeutic option conferring significant survival advantage of both short- and long-term basis. The optimal criteria for selection will also prevent misuse of the liver donor pool as well as to prevent mortality in salvageable patients. Further research needs to be done to identify subset of patients which are at low risk of recidivism and also cannot be managed with pharmacotherapy alone. We reviewed the current knowledge on role of OLT in patient with acute severe alcoholic hepatitis in the present review.

Auer rods in unusual sites: macrophage indigestion

No abstract available.

In vivo anti-inflammatory activity of Tabernaemontana divaricata leaf extract on male albino mice / 中国天然药物

AIM@#To study the in vivo anti-inflammatory activity of Tabernaemontana divaricata leaf extract on male albino mice.@*METHODS@#Aqueous decoction and methanol leaf extracts were tested for their ability to reduce croton oil-induced edema in the mouse ear after topical application. The methanol leaf extract dose-dependently inhibited the croton oil-induced ear edema in mice (ID50 <500 μg·cm(-2)). A bioassay-guided liquid-liquid fractionation of this methanol extract gave four active fractions: water insoluble (F1), hexane (F2), ethyl acetate (F3) and water (F4).@*RESULTS@#The hexane fraction showed a very high activity (42.1% inhibition at 0.7 μg·cm(-2)) as compared to the control. The other fractions were less active (F1: 56.1% at 506.2 μg·cm(-2); F3: 57.3% at 289.3 μg·cm(-2); and F4: 31.9% for 203.8 μg·cm(-2)) while indomethacin gave 48.8% of inhibition at 90 μg·cm(-2). The activity of F1 and F3 may be at least in part explained by the presence of anti-inflammatory flavonoids, while the activity was not correlated to the tannin contents. No compounds were detected in the most active F2 fraction.@*CONCLUSIONS@#The results give a rational support to the traditional use of T. divaricata in tropical India as anti-inflammatory agent.

Inhibitory control test, critical flicker frequency, and psychometric tests in the diagnosis of minimal hepatic encephalopathy in cirrhosis

Minimal hepatic encephalopathy [MHE] impairs health-related quality of life and driving ability of the patient. We assessed the utility of the inhibitory control test [ICT], critical flicker frequency [CFF], and psychometry in the diagnosis of MHE. Consecutive patients with cirrhosis underwent number connection tests A and B [NCT-A, B], digit symbol test [DST], line tracing test [LTT], serial dot test [SDT], CFF, and ICT at baseline and after four hours. Fifty healthy subjects served as controls for the ICT test. Fifty patients with cirrhosis [43.4 +/- 10.2 yrs, M: F 42:8] underwent psychometric tests [NCT-A [48.3 +/- 17.7 vs. 42.6 +/- 17.3 sec, P = 0.001], NCT-B [85.7 +/- 40.1 vs. 90.2 +/- 37.0 sec, P = 0.18], DST [23.5 +/- 9.3 vs. 23.0 +/- 8.7, P = 0.45], LTT [96.6 +/- 48.2 vs. 96.8 +/- 46.8 sec, P = 0.92], SDT [88.0 +/- 39.5 vs. 83.4 +/- 37.2 sec, P = 0.02]] at baseline and after four hours. Target accuracy of ICT was lower in patients with cirrhosis compared with controls [88.4 +/- 5.6 vs. 95.6 +/- 2.1, P = 0.01], whereas ICT lures were higher [18.3 +/- 4.2 vs. 10.2 +/- 2.8, P = 0.01]. Patients with cirrhosis showed a reduction in lures in the second evaluation compared with the first [18.3 +/- 4.2 vs. 17.1 +/- 4.3, P = 0.003] but no change in target accuracy [88.4 +/- 5.6 vs. 88.4 +/- 5.3, P = 0.97]. Control subjects did not show any change either in lures [10.2 +/- 2.8 vs. 10.3 +/- 2.1, P = 0.65] or target accuracy [95.6 +/- 2.1 vs. 95.5 +/- 2.2, P = 0.82]. The sensitivity and specificity of ICT test for the diagnosis of MHE at lure rate >16.5 was 88.5 and 56%, respectively. CFF in patients with MHE [38.4 +/- 1.8 vs. 38.6 +/- 1.5, P = 0.3] and non MHE [40.6 +/- 2.2 vs. 40.8 +/- 2.2, P = 0.6] did not show any difference after four hours as in controls [41.9 +/- 2.4 vs. 42.1 +/- 2.0, P = 0.3]. Thirty one [31%] patients preferred psychometric tests, 57 [57%] preferred CFF and only 12 [12%] preferred ICT [P = 0.001]. ICT, CFF, and psychometric tests are useful tools to assess MHE, and CFF was preferred by this study cohort

Pharmacognostic Screening, Phytochemical Evaluation and In- Vitro free radical Scavenging Activity of Acacia leucophloea Root

Objective: Pharmacognostic Screening and evaluate the in-vitro free radical scavenging activity of roots Acacia leucophloea. Methods: Pharmacognostic Standardization, Physico-chemical evaluation of the roots of Acacia leucophloea was carried out to determine its macro-and microscopical characters and also some of its quantitative standards. Microscopical studies were done by using trinocular microscope. Microscopically, root showed cork, cortex, stellar region and calcium oxalate crystals. Petroleum ether, ethanol, aqueous extracts of Acacia leucophloea were prepared, with successive extraction in soxhlet apparatus. Each extract was selected to study the free radical scavenging activity by superoxide scavenging assay method. Results: It was found that aqueous extract contained carbohydrates, glycosides amino acids flavonoids, tannins, alkaloids, steroids; ethanolic extract contained glycosides amino acids flavonoids, tannins, alkaloids, steroids. Ethanolic extract of Acacia leucophloea shows maximam inhibition in superoxide scavenging model. Aqueous extract also showed almost similar activity compared to ethanolic extract), while Petroleum ether extract showed poor inhibition of superoxide scavenging activity. Conclusion: The present study on pharmacognostic standardization, physico and phytochemical evaluation of Acacia leucophloea root might be useful to supplement information about its identification parameters assumed significantly in the way of acceptability of herbal drugs in present scenario lacking regulatory laws to control quality of herbal drugs.

Minimal hepatic encephalopathy in patients with cirrhosis by measuring liver stiffness and hepatic venous pressure gradient

Transient elastography [TE] of liver and hepatic venous pressure gradient [HVPG] allows accurate prediction of cirrhosis and its complications in patients with chronic liver disease. There is no study on prediction of minimal hepatic encephalopathy [MHE] using TE and HVPG in patients with cirrhosis. Consecutive cirrhotic patients who never had an episode of hepatic encephalopathy [HE] were enrolled. All patients were assessed by psychometry [number connection test [NCT-A and B], digit symbol test [DST], serial dot test [SDT], line tracing test [LTT]], critical flicker frequency test [CFF], TE by FibroScan and HVPG. MHE was diagnosed if there were two or more abnormal psychometry tests [ +/- 2 SD controls]. 150 patients with cirrhosis who underwent HVPG were screened; 91 patients [61%, age 44.0 +/- 11.4 years, M:F:75:16, Child's A:B:C 18:54:19] met the inclusion criteria. Fifty three [58%] patients had MHE [Child A [7/18, 39%], Child B [32/54, 59%] and Child C [14/19, 74%]]. There was no significant difference between alanine aminotranferease [ALT], aspartate aminotransferase [AST] and total bilirubin level in patients with MHE versus non MHE. Patients with MHE had significantly lower CFF than non MHE patients [38.4 +/- 3.0 vs. 40.2 +/- 2.2 Hz, P = 0.002]. TE and HVPG in patients with MHE did not significantly differ from patients with no MHE [30.9 +/- 17.2 vs. 29.8 +/- 18.2 KPas, P = 0.78; and 13.6 +/- 2.7 vs. 13.6 +/- 3.2 mmHg, P = 0.90, respectively].There was significant correlation of TE with Child's score [0.25, P = 0.01], MELD [0.40, P = 0.001] and HVPG [0.72, P = 0.001] while no correlation with psychometric tests, CFF and MHE. TE by FibroScan and HVPG cannot predict minimal hepatic encephalopathy in patients with cirrhosis.

Lactulose for minimal hepatic encephalopathy in patients with extrahepatic portal vein obstruction

Minimal hepatic encephalopathy [MHE] is common in patients with extrahepatic portal vein obstruction [EHPVO]. There is no study on the treatment of MHE using lactulose in patients with EHPVO. Consecutive EHPVO patients were assessed by psychometric [number connection test [NCT-A and B], digit symbol test [DST], serial dot test [SDT], line tracing test [LTT]], and critical flicker frequency [CFF] at inclusion. Patients diagnosed as MHE were treated with lactulose and psychometric tests, CFF, and were reassessed after 3 months. Of the 70 patients screened, the prevalence of abnormal psychometric test was as follows: NCT-A [41%], NCT-B [53%], DST [38%], SDT [40%], and LTT [44%]. Thirty patients [43%] had two or more than two abnormal [>2 SD] psychometry tests. Lactulose improved MHE in 16/30 [53%] of patients after 3 months of treatment. Arterial ammonia decreased after lactulose treatment compared to baseline [83.7 +/- 19.1 vs. 65.1 +/- 19.3 micro mol/l, P=0.001]. A total of 9 [75%] of 12 patients with large spontaneous shunt and 7 [39%] of 18 patients without spontaneous shunt improved with lactulose [P=0.07]. CFF in patients with MHE [n=30] was significantly lower than those without MHE [n=40] [38.1 +/- 2.4 vs. 41.5 +/- 3.1 Hz, P=0.01]. CFF was less than 38 Hz in 21 [70%] of 30 patients before treatment and in 10 [33%] patients after lactulose therapy in MHE patients. All patients could tolerate lactulose without any significant side effects. Four patients [13%] developed transient diarrhea in whom dose needed reduction, 3 [10%] did not like its taste but have continued, and 2 [6%] developed abdominal bloating sensation. Lactulose is effective in the treatment of MHE in patients with EHPVO

Profile of hepatic encephalopathy in children with cirrhosis and response to lactulose

Hepatic encephalopathy [HE] is associated with a poor prognosis. There is paucity of data on the treatment of HE with lactulose in children with cirrhosis. Patients and Retrospective analysis of consecutive cirrhotic patients [<18 years] with HE was done. HE was defined according to West-Haven criteria. Response was defined as complete if patients recovered completely from HE, partial response was defined as improvement of encephalopathy by one or more grades from admission but not complete recovery, and defined as non response if patient did not show any improvement or deteriorated further even after 10 days of lactulose therapy. A total of 300 patients were admitted with cirrhosis and HE [278 adults and 22 children]. Of 22 patients, 16 [73%] patients had complete response to lactulose and six [27%] patients did not [three [13.5%] patients worsened [non response] and three [13.5%] did not recover fully even after 10 days of treatment [partial response]. Comparing baseline characteristics of patients who had complete response [n=16] versus partial [n=3] and non response [w=3], there was significant difference in mean arterial pressure [78.1 +/- 10.7 vs 62.6 +/- 5.0 mmHg, P=0.003], serum sodium [131.3 +/- 3.2 vs 126.515.2, P=0.01] and serum creatinine [0.78 +/- 0.3 vs 1.1 +/- 0.3 mg/dl, P=0.02]. We did not find any difference in baseline characteristics of these patients regarding CTP score [9.6 +/- 1.2 vs 10.6 +/- 1.2], MELD score [17.612.9 vs 17.1 +/- 3.4] severity of HE [2.5 +/- 0.6 vs 2.6 +/- 0.5] and etiology of precipitating factors [P=0.78]. Lactulose therapy causes complete recovery from hepatic encephalopathy in 73% of pediatrics patients with cirrhosis

Predictors of minimal hepatic encephalopathy in patients with cirrhosis

Minimal hepatic encephalopathy [MHE] impairs patient's daily functioning of life. Predictors of MHE in cirrhotic patients have not been evaluated. A total of 200 cirrhotic patients [Child A, 74[37%]; Child B, 72[36%]; Child C, 54[27%]] were evaluated by psychometry, P300 auditory event-related potential [P300ERP] and critical flicker frequency [CFF]. MHE was diagnosed by abnormal psychometry [>2S.D.] and P300ERP [>2.5 S.D.]. Univariate and multivariate logistic regression analyses were performed to determine the predictors of MHE. Eighty-two [41%] patients were diagnosed to have MHE - 26/74 [35%] in Child a, 26/72 [36%] in Child B and 30/54 [56%] in Child C. Ninety-seven [48.5%] patients had abnormal psychometric tests, and 96 [48%] had prolonged P300ERP [>385 ms]. Sixteen [16.5%] patients with abnormal psychometry had P300ERP > 358 ms. One hundred and three patients had CFF value <39 Hz with specificity of 86.6% and sensitivity of 72.9% for MHE. Model for end-stage liver disease [MELD] [17.9 +/- 5.7 vs. 13.4 +/- 4.2, P=0.005], Child-Turcotte-Pugh [CTP] score [8.4 +/- 2.5 vs. 7.7 +/- 2.2, P=0.02], ammonia [104.8 +/- 37.9 vs. 72.5 +/- 45.2 micro mol/L, P=0.001] and CFF [37.0 +/- 2.8 vs. 41.0 +/- 3.4 Hz, P=0.001] were significantly higher in MHE as compared to non-MHE patients. Ninety-one [45.5%] patients had MELD>15.5, 115 [57.5%] had CTP score> 7.5, while 93 [46.5%] had venous ammonia >84.5 micro mol/L. On univariate analysis, MELD [8.52[95% CI, 4.11-16.26; P=0.001]], CFF [17.34[95% CI, 8.16-36.85; P=0.001]] and venous ammonia [7.80 [95% CI, 4.11-14.81; P= 0.003]] were associated with MHE; while CTP score [1.51[95% CI, 0.85-2.69; P=0.30]] was not significant. On multivariate analysis, MELD, CFF and venous ammonia were predictive of MHE. Prevalence of MHE in this study was 41%; and MELD>15.5, CFF< 39 Hz and venous ammonia >84.5 micro mol/L were predictive of MHE