Intestinal protozoan infections among Egyptian neutropenic patients with acute leukemia.

Several enteric protozoan species are linked to diarrhea in humans, with some causing debilitating illnesses, essentially in immunocompromised and neutropenic patients as in acute leukemias. The aim of this study was to detect intestinal protozoa in Egyptian neutropenic patients with acute leukemia. The study comprised two groups; 40 newly diagnosed neutropenic acute leukemia patients and 30 controls. Stool samples were collected from all participants and subjected to routine microscopic examination, special staining and detection of copro-antigen using rapid diagnostic test (RDT) RIDA®QUICK Entamoeba/ Giardia/ Cryptosporidium Combi. Cases were tested post-chemotherapy at the nadir of neutropenia (absolute neutrophil count ANC<= 0.5x109/L) and 19 cases were also tested initially prior to chemotherapy. Of examined patients, 15/40 (37%) were positive for Blastocystis hominis by wet mount, 10/40 (25%) had microsporidia using modified trichrome stain and only 2 cases (5%) of Cryptosporidium spp. by Ziehl-Neelsen stain. By RDT, 8/40 cases (20%) were positive compared to entirely negative controls. The positive cases included 4 patients with G. intestinalis 2 with Entamoeba and 2 with Cryptosporidium.19/40 cases were tested both pre- and post-chemotherapy. microsporidian spp. was diagnosed in 6/19 cases at the nadir of neutropenia compared to none of the cases pre-chemotherapy and the difference was statistically significant (p= 0.031*). Intestinal protozoa in acute leukemia patients post-chemotherapy are common especially B. hominis. Furthermore, RDT might be helpful for diagnosing intestinal protozoa in acute leukemia. Attention is highly required as intestinal protozoa infection can emerge after chemotherapy such as microsporidia.

Intestinal protozoan infections among Egyptian neutropenic patients with acute leukemia

@#Several enteric protozoan species are linked to diarrhea in humans, with some causing debilitating illnesses, essentially in immunocompromised and neutropenic patients as in acute leukemias. The aim of this study was to detect intestinal protozoa in Egyptian neutropenic patients with acute leukemia. The study comprised two groups; 40 newly diagnosed neutropenic acute leukemia patients and 30 controls. Stool samples were collected from all participants and subjected to routine microscopic examination, special staining and detection of copro-antigen using rapid diagnostic test (RDT) RIDA®QUICK Entamoeba/ Giardia/ Cryptosporidium Combi. Cases were tested post-chemotherapy at the nadir of neutropenia (absolute neutrophil count ANC< 0.5x109/L) and 19 cases were also tested initially prior to chemotherapy. Of examined patients, 15/40 (37%) were positive for Blastocystis hominis by wet mount, 10/40 (25%) had microsporidia using modified trichrome stain and only 2 cases (5%) of Cryptosporidium spp. by Ziehl-Neelsen stain. By RDT, 8/40 cases (20%) were positive compared to entirely negative controls. The positive cases included 4 patients with G. intestinalis 2 with Entamoeba and 2 with Cryptosporidium.19/40 cases were tested both pre- and post-chemotherapy. microsporidian spp. was diagnosed in 6/19 cases at the nadir of neutropenia compared to none of the cases pre-chemotherapy and the difference was statistically significant (p= 0.031*). Intestinal protozoa in acute leukemia patients post-chemotherapy are common especially B. hominis. Furthermore, RDT might be helpful for diagnosing intestinal protozoa in acute leukemia. Attention is highly required as intestinal protozoa infection can emerge after chemotherapy such as microsporidia.

Interleukins 12 and 13 levels among beta-thalassaemia major patients.

The role of inflammatory cytokines in the pathophysiology of beta-thalassaemia is still unclear. In this study production levels of interleukins (IL)-12 and IL-13 were measured by commercial ELISA in culture supernatants of mitogen-stimulated peripheral blood mononuclear cells-from 30 non-splenectomized beta-thalassaemia cases with iron overload and 20 age- and sex-matched healthy individuals. IL-12 levels were significantly lower among cases compared with controls (91.4 pg/mL versus 154.6 pg/mL), while IL-13 levels were significantly higher (42.5 pg/mL versus 5.7 pg/mL). There was a significant negative correlation between IL-12 and lL-13 levels among beta-thalassaemia cases (r= -0.42). Patients with beta-thalassaemia alone had higher IL-12 levels than beta-thalassaemia patients who were seropositive for chronic hepatitis B or C virus infection (140 pg/mL versus 50 pg/mL); IL-13 levels were slightly lower (65 pg/mL versus 67 pg/mL). An imbalance in the IL-12/IL-13 axis may be relevant to the pathophysiology of beta-thalassaemia.

Quantitative estimation of interferon-gamma levels among Egyptian polytransfused haematology cases.

This study was designed to estimate interferon-gamma (INF-gamma) levels among polytransfused haematology cases. Cases were selected from the haematology unit of Alexandria main university hospital, Egypt. Complete blood counts, estimation of INF-gamma and hepatitis B and C virus (HBV and HCV) status were conducted on 20 unsplenectomized patients with gamma-thalassaemia major and 20 patients with acute myeloid leukaemia (AML) in the maintenance phase and 20 healthy subjects. Mean haemoglobin levels and red blood cell counts were significantly higher in the control group than the AML and thalassaemia groups, while white blood cell counts were significantly lower in the control group than the case groups. Two AML patients (10%) and 1 thalassaemia patient (5%) were HBV-positive, while 5% of both case groups were HCV-positive. Mean values of INF-gamma were significantly different between AML, thalassaemia major and control groups: 5517 (SD 1142) pg/mL, 1024 (SD 249) pg/mL and 2980 (SD 604) pg/mL respectively.

Study of host immunity in patients with Helicobacter pylori-related idiopathic thrombocytopenia.

To test the role of T helper cell Th1 immunity we recruited 24 patients with idiopathic thrombocytopenia associated with H. pylori seropositivity. They were divided into 2 groups: 12 with immune-mediated thrombocytopenia (Group 1) and 12 with non-immune mediated thrombocytopenia (Group 2). We also recruited 10 individuals seronegative for H. pylori (Group 3) as controls. Initial platelet count was significantly lower in Group 1 than Group 2. H. pylori was eradicated in 10 of 12 patients in Group 1 and in all patients in Group 2. Transient improvement (< 3 months) in the platelet count occurred in only 2 patients in Group 1 while improvement for > 6 months was observed in all patients in Group 2. There was a statistically significant direct correlation between platelet count and levels of TNF- and IFN-gamma in both study groups, while a non-significant correlation was seen in Group 3. Thus, H. pylori infection should be considered in the differential diagnosis of all cases of thrombocytopenia, and should be eradicated in all H. pylori-positive patients with thrombocytopenia.

Interleukins 12 and 13 levels among beta-thalassaemia major patients

The role of inflammatory cytokines in the pathophysiology of beta-thalassaemia is still unclear. In this study production levels of interleukins [IL]-12 and IL-13 were measured by commercial ELISA in cultureI supernatants of mitogen-stimulated peripheral blood mononuclear cells from 30 non-splenectomized beta-thalassaemia cases with iron overload and 20 age- and sex-matched healthy individuals. IL-12 levels were significantly lower among cases compared with controls [91.4 pg/mL versus 154.6 pg/mL] while IL-13 levels were significantly higher [42.5 pg/mLversus 5.7 pg/mL]. There was a significant negative correlation between IL-12 and IL-13 levels among beta-thalassaemia cases [r= -0.42]. Patients with beta-thalassaemia alone had higher IL-12 levels than beta-thalassaemia patients who were seropositive for chronic hepatitis B or C virus Infection [140 pg/mL versus 50 pg/mL]; IL-13 levels were slightly lower [65 pg/mL versus 67 pg/mL]. An imbalance In the IL-12/IL-13 axis may be relevant to the pathophysiology of beta-thalassaemia

Quantitative estimation of interferon-gamma levels among Egyptian polytransfused haematology cases

This study was designed to estimate interferon-gamma [INF-gamma] levels among polytransfused haematology cases. Cases were selected from the haematology unit of Alexandria main university hospital, Egypt. Complete blood counts, estimation of INF-gamma and hepatitis B and C virus [HBV and HCV] status were conducted on 20 unsplenectomized patients with thalassaemia major and 20 patients with acute myeloid leukaemia [AML] in the maintenance phase and 20 healthy subjects. Mean haemoglobin levels and red blood cell counts were significantly higher in the control group than the AML and thalassaemia groups, while white blood cell counts were significantly lower in the control group than the case groups. Two AML patients [10%] and 1 thalassaemia patient [5%] were HBV-positive, while 5% of both case groups were HCV-positive. Mean values of INF-? were significantly different between AML, thalassaemia major and control groups: 5517 [SD 1142] pg/mL, 1024 [SD 249] pg/mL and 2980 [SD 604] pg/mL respectively

Study of host immunity in patients with Helicobacter pylori-related idiopathic thrombocytopenia

To test the role of T helper cell Th1 immunity we recruited 24 patients with idiopathic thrombocytopenia associated with H. pylori seropositivity. They were divided into 2 groups: 12 with immune-mediated thrombocytopenia [Group 1] and 12 with non-immune mediated thrombocytopenia [Group 2]. We also recruited 10 individuals seronegative for H. pylori [Group 3] as controls. Initial platelet count was significantly lower in Group 1 than Group 2. H. pyloriwas eradicated in 10 of 12 patients in Group 1 and in all patients in Group 2. Transient improvement [< 3 months] in the platelet count occurred in only 2 patients in Group 1 while improvement for > 6 months was observed in all patients in Group 2. There was a statistically significant direct correlation between platelet count and levels of TNF- and IFN-gamma in both study groups, while a non-significant correlation was seen in Group 3. Thus, H. pyloriinfection should be considered in the differential diagnosis of all cases of thrombocytopenia, and should be eradicated in all H. pylori-positive patients with thrombocytopenia

Study of anticardiolipin antibody in hepatitis C virus-positive patients

Several antibodies, including anticardiolipin antibodies (ACA), have been detected among chronically infected hepatitis C virus (HCV) patients. The present work aimed at ascertaining the clinical significance of ACA levels among HCV infection associated with two commonly encountered diseases, thrombocytopenia and arteriovenous-shunt malfunction. Six groups were studied, 11 HCV-positive thrombocytopenic patients (group I), 14 HCV-positive non-thrombocytopenic patients (group II) and 15 healthy controls (group III), 11 anti-HCV-positive hemodialysis patients with non-functioning shunt (group IV), 14 anti-HCV-positive hemodialysis patients with patent shunt (group V) (Bain Medical Equipment Co., China) and 15 healthy controls (group VI). Anticardiolipin antibody (ACA) assay was performed on all patients and controls whereas tumor necrosis factor α (TNF-α) assay was carried out on thrombocytopenic patients and controls. Thrombocytopenic groups presented an inverse correlation between IgG ACA levels and both thrombocytopenia and TNF-α levels. During the follow-up period, no other clinical manifestations related to ACA were developed. Hemodialysis groups showed a significant elevation in IgG ACA levels in groups IV and V compared to the controls, with statistically higher levels in group IV than group V. Three group IV patients were hypercholesterolemic. We can conclude that induction of proinflammatory cytokines such as TNF-α by persistent HCV infection may promote the generation of ACA. Complications of HCV, including thrombocytopenia and thrombosis in arteriovenous shunt, are more strongly correlated with IgG ACA than with IgM ACA.(AU)

Levels of interleukins 12 (IL-12) and 13 (IL-13), hepatitis B and C serology, and blood cultures among acute myeloid leukemia (AML) patients in Egypt

There is an interest in the use of IL-12 as a possible anti-cancer drug to induce immune responses and anti-IL-13 formulations to treat the undesirable effects of IL-13. Thus, the present study aimed at analyzing IL-12 and IL-13 profiles, viral hepatitis serology and blood cultures in acute myeloid leukemia (AML) patients. Forty individuals (20 without septicemia - Group A, and 20 with septicemia - Group B) and 20 healthy controls were evaluated. Hepatitis B virus antigens (HBsAg) and hepatitis C virus antibodies (HCV Ab) were quantified using commercial ELISA kits. IL-12 and IL-13 levels were estimated in culture supernatant of mitogen-stimulated peripheral blood mononuclear cells by ELISA. Significantly low IL-12 values were observed among AML patients compared to controls whereas the opposite was observed regarding IL-13. IL-12 levels were found to be elevated in the follow-up cases. M4 and M5 subtypes of AML presented higher IL-12 levels than M1 and M2 subtypes. The isolated organisms from AML with septicemia were Staphylococcus aureus (35 percent), Esherichia coli (25 percent), coagulase-negative staphylococci (25 percent), and Candida (15 percent). Fungemia cases showed higher IL-12 values than bacteremia cases. In conclusion, IL-12 and IL-13 should be further tested in large-scale studies to provide future immunotherapy against AML.(AU)

Correlation between serum levels of interleukins 10 and 12 and thrombocytopenia in hepatitis C cirrhotic (class A) patients

Hepatitis C virus (HCV) patients commonly have low platelet counts; however, the exact role of HCV infection in thrombocytopenia is unknown. This work aimed to study the serum levels of interleukins (IL) 10 and 12 in patients with mild and moderate thrombocytopenia associated with chronic hepatitis C infection. Our study included 15 patients with chronic HCV infection and newly diagnosed isolated autoimmune thrombocytopenia (Group I) and 15 patients with chronic HCV infection and normal platelet count as controls (Group II). All patients were examined for personal history and clinical aspects, complete blood count, bone marrow aspiration, liver function tests, HCV antibody assay by ELISA and polymerase chain reaction (PCR), abdominal ultrasound, Helicobacter pylori stool antigen test, evaluation of serum levels of IL-10, IL-12 and platelet specific antibodies. Our results revealed that eight patients from Group l had mild thrombocytopenia and seven patients had moderate thrombocytopenia. Serum IL-10 level was significantly elevated (t = 9.301, p < 0.001) while serum IL-12 showed a significant decrease (t = 6.502, p < 0.001) in Group I compared to the control group. No correlation was detected between platelet counts and the serum levels of either IL-10 [r = 0.454, p = 0.089 (Group I), r = 0.038, p = 0.89 (Group II)] or IL-12 [r = 0.497, p = 0.06 (Group I), r = 0.499, p = 0.058 (Group II)]. However, in Group I, a significant correlation was present only between moderate thrombocytopenia and serum levels of either IL-10 (r = 0.794, p = 0.033) or IL-12 (r = 0.967, p = 0.001), while no correlation was detected between these interleukin parameters and mild thrombocytopenia (r = 0.311 and p = 0.453 for IL-10 and r = -0.08 and p = 0.851 for IL-12). Based on our data, we may conclude that interleukins 10 and 12 are involved in low platelet levels.(AU)

Role of Helicobacter pylori in refractory iron deficiency anaemia.

The role of Helicobacter pylori infection in the development of iron deficiency anaemia has been the focus of attention over the past decade. However, confirmation of a relationship has not confirmed the pathophysiological mechanisms involved in the phenomenon. The aim of the present work is to study the levels of fasting gastric acidity (free and total) as well as the level of tumour necrosis factor-alpha (TNF alpha) in male refractory iron deficiency anaemia patients seropositive for H. pylori infection versus those who are seronegative. Thirty adult patients with iron deficiency anaemia and gastroduodenitis were subdivided into two groups of matched age and haemoglobin value. Group 1 was H. pylori-seropositive for infection and these patients did not receive prior treatment for eradication of H. pylori infection. Group 2 comprised patients seronegative for H. pylori infection (control group). Patients with active bleeding or previous medical problems were excluded from the study. All patients and controls were subjected to the following at presentation: history taking and thorough clinical examination, complete blood picture, reticulocytes (%), assessment of serum iron, total iron binding capacity, serum ferritin, IgG anti-Helicobacter antibody and TNF alpha, stool for occult blood and measurement of gastric acidity (total and free). Upper endoscopy was performed and multiple biopsies were taken and tested for expression of cytotoxin-associated gene A (cagA) by the polymerase chain reaction (PCR). Results showed significantly higher values of free and total gastric acidity as well as TNF alpha levels in Group 1 compared to controls (Group 2). Among those in Group 1, higher TNF alpha levels were seen in seven H. pylori cagA-positive patients than in eight cagA-negative patients. Haemoglobin values were inversely correlated with TNF alpha levels. Thus, elevated serum TNF alpha in the H. pylori-seropositive group may be one of the underlying pathophysiological mechanism for iron deficiency anaemia observed in these patients.

Bioconversion of poultry wastes I-factors influencing the assay and productivity of crude uricase by three uricolytic filamentous fungi.

The optimum temperature for biomass yield and uricase production by uricolytic fungi, Aspergillus terreus. A. flavus and Trichoderma sp. was at 30 degrees C. The time required for maximum production of uricase and biomass yield was 4 days for two Aspergillus species and 6 days for Trichoderma sp. The optimum pH was at 6.4 for A. terreus and pH 6.6 for both A. flavus and Trichoderma sp. The maximum fungal biomass yield was achieved in medium supplemented with 4% poultry waste. The best carbon sources for the production of uricase and mycelia yield were glycerol, sucrose and maltose by A. terreus, A. flavus and Trichoderma sp., respectively. Uric acid was found to be the best nitrogen source for production and activity of uricase by the three tested fungi. The addition of some vitamins to the culture media increased the maximum biomass yield of all the isolates, but did not significantly increase uricase production.

Bioconversion of poultry wastes. I--Factors influencing the assay and productivity of crude uricase by three uricolytic filamentous fungi.

The optimum temperature for biomass yield and uricase production by uricolytic fungi, Aspergillus terreus, A. flavus and Trichoderma sp. was at 30 degrees C. The time required for maximum production of uricase and biomass yield was 4 days for two Aspergillus species and 6 days for Trichoderma sp. The optimum pH was at 6.4 for A. terreus and pH 6.6 for A. flavus and Trichoderma sp. The maximum fungal biomass yield was achieved in medium supplemented with 4% poultry waste. The best carbon sources for the production of uricase and mycelia yield were glycerol, sucrose and maltose by A. terreus, A. flavus and Trichoderma sp., respectively. Uric acid was found to be the best nitrogen source for production and activity of uricase by the three tested fungi. The addition of some vitamins to the culture media increased the maximum biomass yield of all the isolates, although no significantly increased uricase production was found.