Clinical perspectives, immunohematologic insights, and transfusion management in IgA-associated autoimmune hemolytic anemia.

Autoimmune hemolytic anemia (AIHA) due to warm-reacting IgA autoantibodies is rare. Here, we explored the clinical and immunohematologic characteristics of patients suffering from IgA-associated warm AIHA (WAIHA) and their transfusion management. The 9-year study included 214 patients with WAIHA who were further classified into two groups: (1) IgA-associated WAIHA and (2) non-IgA-associated WAIHA. Clinical and laboratory details were obtained from patient files and the Hospital Information System. All immunohematologic investigations were performed following standard operating procedures and established protocols. Among the 214 patients with WAIHA, 17 (7.9%) belonged to the IgA-associated group; of these, two IgA-only WAIHA cases were found. The mean hemoglobin in this group was 5.58 g/dL, and 15 (88.2%) of these patients received a total of 32 units of packed red blood cell (RBC) transfusions. In vivo hemolytic markers were significantly abnormal in the IgA-associated WAIHA group when compared with the non-IgA group. Secondary WAIHA was found in 11 (64.7%) patients with IgA-associated WAIHA. Patients with IgA-associated WAIHA received more blood transfusions than individuals in the non-IgA group (p = 0.0004). A total of 17 (7.9%) patients with WAIHA experienced adverse events to blood transfusion. Detailed characterization of WAIHA with particular emphasis on IgA-associated and non-IgA-associated WAIHA is essential to evaluate the disease characteristics, access the degree of hemolysis, understand the immunohematologic behaviors of the antibodies, and manage blood transfusions.

Occurrence of Schistosoma nasale infection in bullocks of Puducherry.

Nasal schistosomiasis is caused by the blood fluke Schistosoma nasale (S. nasalis) adversely affects the health and production of domestic livestock in various parts of India. The present report describes the occurrence of S. nasale infection in two Hallikar breed bullocks of Union Territory of Puducherry. Eggs of S. nasale were noticed in nasal washings/scrapings of animals and identified as per the standard taxonomical keys.

Composition, in vitro antioxidant and antimicrobial activities of essential oil and oleoresins obtained from black cumin seeds (Nigella sativa L.).

Gas chromatography-mass spectrometry (GC-MS) analysis revealed the major components in black cumin essential oils which were thymoquinone (37.6%) followed by p-cymene (31.2%), α-thujene (5.6%), thymohydroquinone (3.4%), and longifolene (2.0%), whereas the oleoresins extracted in different solvents contain linoleic acid as a major component. The antioxidant activity of essential oil and oleoresins was evaluated against linseed oil system at 200 ppm concentration by peroxide value, thiobarbituric acid value, ferric thiocyanate, ferrous ion chelating activity, and 1,1-diphenyl-2-picrylhydrazyl radical scavenging methods. The essential oil and ethyl acetate oleoresin were found to be better than synthetic antioxidants. The total phenol contents (gallic acid equivalents, mg GAE per g) in black cumin essential oil, ethyl acetate, ethanol, and n-hexane oleoresins were calculated as 11.47 ± 0.05, 10.88 ± 0.9, 9.68 ± 0.06, and 8.33 ± 0.01, respectively, by Folin-Ciocalteau method. The essential oil showed up to 90% zone inhibition against Fusarium moniliforme in inverted petri plate method. Using agar well diffusion method for evaluating antibacterial activity, the essential oil was found to be highly effective against Gram-positive bacteria.

Determination of anthelmintic activity of the leaf and bark extract of tamarindus indica linn.

The aim of the present study was to evaluate the anthelmintic activity of ethanolic and aqueous extract of leaves and bark of Tamarindus indica Linn using Pheretima posthuma and Tubifex tubifex as test worms. The time of paralysis and time of death were studied and the activity was compared with piperazine citrate as reference standard. The alcohol and aqueous extract of bark of Tamarindus indica exhibited significant anthelmintic activity as evidenced by decreased paralyzing time and death time. The results thus support the use of Tamarindus indica as an anthelmintic agent.

Effects of Petroleum Ether Extract of Amorphophallus paeoniifolius Tuber on Central Nervous System in Mice.

The central nervous system activity of the petroleum ether extract of Amorphophallus paeoniifolius tuber was examined in mice, fed normal as well as healthy conditions. The petroleum ether extract of Amorphophallus paeoniifolius tuber at the doses of 100, 300 and 1000 mg/kg showed significant central nervous system activity in mice.

Application of gel technology in the serologic characterization of autoantibody in DAT-positive autoimmune diseases.

Gel tests are now available for the determination of immunoglobulin classes and subclasses and complement fractions coating RBCs. These tests simplified serologic characterization of autoantibodies in various autoimmune diseases. The aim of this study was to evaluate the use of gel cards in the serologic characterization of autoantibody with regard to the immunoglobulin classes, complement fractions, and IgG subclasses, and the influence of these characteristics on hemolysis. Gel cards were used to characterize the RBC-bound autoantibodies in 66 DAT-positive patients. Hematologic and biochemical parameters such as Hb, reticulocyte count, serum bilirubin, and serum LDH were obtained from the patient files. Of the 49 patients carrying IgG on their RBCs, 21 (42.8%) were suffering from connective tissue disorders and another 16 patients (32.7%) had autoimmune hemolytic anemia. A total of 19 of these 49 patients had evidence of hemolysis. Thirteen of the 17 patients (76.5%) whose RBCs were coated with more than one type of immunoglobulin and complement were experiencing hemolysis (p < 0.05). Seventy-five percent (21 of 28) of patients having IgG1, IgG3, or both on their RBCs showed hemolysis (p < 0.05). Thus, it is important to serologically characterize autoantibodies in autoimmune disorders to effectively predict the prognosis and disease outcome. This characterization can be performed effectively with the gel test, which can be introduced in blood centers as a replacement to the conventional tube technique.

Enterotoxigenicity of mature 45-kilodalton and processed 35-kilodalton forms of hemagglutinin protease purified from a cholera toxin gene-negative Vibrio cholerae non-O1, non-O139 strain.

Cholera toxin gene-negative Vibrio cholerae non-O1, non-O139 strain PL-21 is the etiologic agent of cholera-like syndrome. Hemagglutinin protease (HAP) is one of the major secretory proteins of PL-21. The mature 45-kDa and processed 35-kDa forms of HAP were purified in the presence and absence of EDTA from culture supernatants of PL-21. Enterotoxigenicities of both forms of HAP were tested in rabbit ileal loop (RIL), Ussing chamber, and tissue culture assays. The 35-kDa HAP showed hemorrhagic fluid response in a dose-dependent manner in the RIL assay. Histopathological examination of 20 microg of purified protease-treated rabbit ileum showed the presence of erythrocytes and neutrophils in the upper part of the villous lamina propria. Treatment with 40 microg of protease resulted in gross damage of the villous epithelium with inflammation, hemorrhage, and necrosis. The 35-kDa form of HAP, when added to the lumenal surface of rat ileum loaded in an Ussing chamber, showed a decrease in the intestinal short-circuit current and a cell rounding effect on HeLa cells. The mature 45-kDa form of HAP showed an increase in intestinal short-circuit current in an Ussing chamber and a cell distending effect on HeLa cells. These results show that HAP may play a role in the pathogenesis of PL-21.

Treatment of prosthetic valve infective endocarditis due to multi-resistant Gram-positive bacteria with linezolid.

OBJECTIVES: Clinical experience with linezolid in the treatment of infective endocarditis either alone or in combination with other agents is limited. We describe our experience in the treatment of two patients with IE due to multi-resistant Gram-positive bacteria. METHODS: One patient with MRSE and one with VRE endocarditis were treated with regimens containing linezolid. The killing kinetics of linezolid in combination with gentamicin or vancomycin against isolates of Staphylococcus epidermidis and Enterococcus faecalis were analysed in vitro. RESULTS: Clinical response and eradication of bacteraemia was achieved with linezolid therapy in both patients. Time-kill curve studies showed that linezolid was bacteriostatic against the MRSE and VRE isolates used. Combination with gentamicin or vancomycin did not produce synergy or antagonism but at best only marginal additive effect. CONCLUSIONS: Although bacteriostatic, linezolid provides an important therapeutic option in IE due to multi-resistant Gram-positive pathogens. It challenges the conventional wisdom that bactericidal synergy is required for the effective treatment of most cases of IE due to Gram-positive organisms.

Factors influencing yield of plateletpheresis using intermittent flow cell separator.

Platelet recovery in the recipient is influenced by the transfused dose of platelets, which in turn is dependent on the quality of single donor platelets (SDPs) in terms of platelet yield. Various donor factors such as predonation platelet count and Hemoglobin (Hb) concentration affect the platelet yield. A total of 61 plateletpheresis procedures performed on intermittent flow cell separator (MCS3p, Hemonetics) were evaluated for platelet yield. A relationship between predonation platelet count and Hb concentration with yield of platelets was studied using Pearson Correlation. The mean platelet yield was 2.9 +/- 0.64 x 10(11). While a direct relationship was observed between predonation platelet count and yield (r = 0.51, P < 0.001), no such correlation was noticed with donor Hb concentration (r = -0.05, P > 0.005). The yield was > or =3 x 10(11) in >80% of procedures when the predonation platelet count was > or =250 x 10(3)/mm. Optimization of platelet yield, which is influenced by predonation platelet count, is an emerging issue in blood transfusion services. However, further studies in this regard are needed using more advanced cell separators.

Calcium and magnesium levels during automated plateletpheresis in normal donors.

It is well known that citrate induces ionized hypocalcaemia by the chelating effect during plateletpheresis. However, the kinetics of serum magnesium (Mg) ions has not been well documented. We, therefore, evaluated biochemical changes in healthy donors during plateletpheresis procedure. Ten healthy donors underwent plateletpheresis on continuous cell separator (CS3000, Baxter, Round Lake, IL, USA) and 10 on intermittent flow cell separator (MCS 3p, Hemonetics, Braintree, MA, USA). Serum levels of total and ionized calcium (tCa and iCa, respectively) and Mg (tMg and iMg, respectively) were measured before, during and after the procedures. Although, the fall in tCa (from 2.62 +/- 0.12 to 2.36 +/- 0.12 mmol L(-1)) and tMg (from 0.89 +/- 0.01 to 0.79 +/- 0.01 mmol L(-1)) was modest and not significant; drop in iCa (from 1.33 +/- 0.1 to 0.84 +/- 0.1 mmol L(-1)) and iMg (from 0.53 +/- 0.01 to 0.35 +/- 0.1 mmol L(-1)) was statistically significant (P < 0.001). There were no significant differences observed between the CS3000 and MCS 3p cell separators regarding the fall in Ca and Mg. None of the donors experienced any adverse reactions during the procedures. In the study, an acute ionized hypocalcaemia and hypomagnesaemia have been observed after the plateletpheresis; therefore, measurement of both the ions may be monitored. However, there is no justification for prophylactic supplementation of either of these elements.

Dietary yeast RNA supplementation reduces mortality by Aeromonas hydrophila in rohu (Labeo rohita L.) juveniles.

A feeding trial was conducted for 60 days to delineate the effect of dietary ribonucleic acid or chitin on haematological parameters, phagocyte respiratory burst and resistance to Aeromonas hydrophila of Labeo rohita juveniles. One hundred and twenty-six (avg. wt. 13.40 +/- 0.17 g) juveniles were randomly distributed in six treatment groups, each one in three replicates. Six isonitrogenous (crude protein: 34.34-35.37%) and isocaloric (414-425 kcal 100 g(-1)) purified diets were prepared with different concentrations of either ribonucleic acid or chitin except the control group, viz., control, T1 (0.1% ribonucleic acid), T2 (0.2% ribonucleic acid), T3 (0.4% ribonucleic acid), T4 (25 mg chitin kg (-1)) and T5 (50 mg chitin kg (-1)). Weight gain %, specific growth rate, feed efficiency ratio, protein efficiency ratio did not vary significantly (P > 0.05) among the experimental groups. Haemoglobin content and total erythrocyte count were observed within the normal range and were not influenced by the dietary immunostimulants. Highest total leukocyte count was found in the T(3) group. The immunomodulatory effects of dietary immunostimulants were studied by using nitroblue tetrazolium (NBT) assay and serum parameters, namely total protein, albumin, globulin and A/G ratio. The respiratory burst activity (NBT) of blood phagocytes was highest in the T3 group followed by the T2 group, which varied significantly (P < 0.05) from other groups. Significantly (P < 0.05) higher total protein, globulin and lower A/G ratio was observed in the T(3) group. The relative percent survival after challenging with Aeromonas hydrophila was highest in the T3 group, compared to the control group, followed by the T2 group. The results indicate that dietary ribonucleic acid at 0.4% enhances phagocyte respiratory burst and protection of Labeo rohita juveniles to challenge by A. hydrophila.

Imaging bacterial infection with (99m)Tc-ciprofloxacin (Infecton).

AIMS: The diagnosis of deep seated bacterial infections, such as intra-abdominal abscesses, endocarditis, and osteomyelitis, can be difficult and delayed, thereby compromising effective treatment. This study assessed the efficacy of a new radioimaging agent, Tc-99m ciprofloxacin (Infecton), in accurately detecting sites of bacterial infection. METHODS: Eight hundred and seventy nine patients with suspected bacterial infection underwent Infecton imaging and microbiological evaluation. The sensitivity and specificity of Infecton in detecting sites of bacterial infection were determined with respect to Centres of Disease Control, World Health Organisation, and Dukes's criteria. RESULTS: Five hundred and seventy four positive and 295 negative images were produced. These included 528 true positives, 46 false positives, 205 true negatives and 90 false negatives, giving an overall sensitivity of 85.4% and a specificity of 81.7% for detecting infective foci. Sensitivity was higher (87.6%) in microbiologically confirmed infections. CONCLUSIONS: Infecton is a sensitive technique, which aids in the earlier detection and treatment of a wide variety of deep seated bacterial infections. The ability to localise infective foci accurately is also important for surgical intervention, such as drainage of abscesses. In addition, serial imaging with Infecton might be useful in monitoring clinical response and optimising the duration of antimicrobial treatment.

Not everything acid fast is Mycobacterium tuberculosis--a case report.

The Ziehl-Neelsen (ZN) stain is important in identifying organisms that are acid fast, principally Mycobacterium tuberculosis. However, decolorisation with a weaker acid concentration (for example 1% hydrochloric acid), often used in ZN staining in histology, can result in a wider variety of organisms appearing acid fast and can be a cause of misidentification. To illustrate this point, a patient is described with pulmonary nocardiosis who was misdiagnosed as having tuberculous empyema on pleural biopsy.

Diagnosis and treatment of cervical intraepithelial neoplasia in a single visit.

We evaluated the diagnostic and therapeutic efficacy of large loop excision of the transformation zone (LLETZ) performed in the first visit (see and treat policy) as compared to LLETZ treatment performed as an interval procedure. Data of 248 patients were analyzed of which 206 patients had LLETZ. Two thirds of the procedures were performed at the first visit. Of all the women who had the 'see and treat policy' 94.9% were diagnosed to have cervical intraepithelial neoplasia (CIN) on histology compared to 90.8% in the interval treatment group. A total of 3 (1.4%) patients were detected to have microinvasive carcinoma. There was no difference in the immediate postoperative complication rate, overtreatment rate and need of repeat treatment in both the groups. Single visit colposcopy and loop treatment is a safe and effective option for treatment of cervical epithelial abnormalities. The experience of the colposcopist is very important.

Evaluation of the efficacy of 99mTc-Infecton, a novel agent for detecting sites of infection.

AIMS: To determine the sensitivity and specificity of 99mTc-Infecton (Infecton), a novel ciprofloxacin based imaging agent, in detecting sites of infection. METHODS: Ninety patients thought to be suffering from a variety of infections were administered 300-400 MBq of Infecton intravenously. Whole body images were taken one and four hours later. Appropriate specimens were taken for microbiological investigations. Statistical analysis was performed using a computer statistical package. RESULTS: Ninety eight Infecton images were produced. Forty one of these were positive, including three false positives, where the patients had non-infective conditions. Fifty seven negative images were obtained, of which 41 were true negatives and 16 were false negatives, having definite evidence of infection. Thus, Infecton imaging has a sensitivity of 70.3% and a specificity of 93.1% for detecting infective foci. The positive and negative predictive values were 92.6% and 71.9%, respectively. CONCLUSION: Infecton imaging is a new diagnostic tool that is specific for detecting sites of bacterial infection in the body. The high positive predictive value displayed by the technique is clinically important because a positive image strongly supports a diagnosis of bacterial infection. A negative result does not rule out an infection, and may be a result of previous or current antibiotic treatment and/or infection with organisms that do not take up Infecton. Infecton imaging has major advantages over well established imaging techniques, including radiolabelled leucocytes, and may prove to be a superior method for localising bacterial infections.