Human Umbilical Cord Blood CD34-Positive Cells as Predictors of the Incidence and Short-Term Outcome of Neonatal Hypoxic-Ischemic Encephalopathy: A Pilot Study

BACKGROUND AND PURPOSE: Neonatal hypoxic-ischemic encephalopathy (HIE) is one of the leading causes of neurological handicap in developing countries. Human umbilical cord blood (hUCB) CD34-positive (CD34⁺) stem cells exhibit the potential for neural repair. We tested the hypothesis that hUCB CD34⁺ stem cells and other cell types [leukocytes and nucleated red blood cells (NRBCs)] that are up-regulated during the acute stage of perinatal asphyxia (PA) could play a role in the early prediction of the occurrence, severity, and mortality of HIE. METHODS: This case-control pilot study investigated consecutive neonates exposed to PA. The hUCB CD34⁺ cell count in mononuclear layers was assayed using a flow cytometer. Twenty full-term neonates with PA and 25 healthy neonates were enrolled in the study. RESULTS: The absolute CD34⁺ cell count (p=0.02) and the relative CD34⁺ cell count (CD34+%) (p<0.001) in hUCB were higher in the HIE patients (n=20) than the healthy controls. The hUCB absolute CD34⁺ cell count (p=0.04), CD34⁺% (p<0.01), and Hobel risk scores (p=0.04) were higher in patients with moderate-to-severe HIE (n=9) than in those with mild HIE (n=11). The absolute CD34⁺ cell count was strongly correlated with CD34⁺% (p<0.001), Hobel risk score (p=0.04), total leukocyte count (TLC) (p<0.001), and NRBC count (p=0.01). CD34+% was correlated with TLC (p=0.02). CONCLUSIONS: hUCB CD34⁺ cells can be used to predict the occurrence, severity, and mortality of neonatal HIE after PA.

Nuclear factor KAPPA-B, tumor necrosis factor-alpha and the transforming growth factor-beta as biomarkers of hepatocellular carcinoma on top of HCV infection

The current study was aimed to investigate the usefulness of nuclear factor kappa- B [NF- kB] expression, tumor necrosis factor alpha [TNF- alpha] and the transforming growth factor-beta [TGF-beta1] as markers in prediction and prognosis of hepatocellular carcinoma [HCC] on top of HCV. The current study was performed on 30 male Egyptian patients, their age ranged from 43 to 74. Twenty patients with hepatitis C virus'related hepatocellular carcinoma [HCV-related HCC] with no evidence of extrahepatic metastasis or hepatitis B surface antigen [HBsAg] and 10 Egyptian patients with chronic active hepatitis C virus, with matched age and sex. All patients were infected with HCV genotype-4a. Fasting blood samples were collected from all subjects [10 ml each]. 1 ml was added onto EDTA for immediate extraction of NF- kB RNA. The serum was used for the qualitative determination of HCV antibodies, quantitative determination of HCV-RNA by PCR, quantitative determination of the bio-markers [AFP-TNF-alpha and TGF-beta1], as well as quantitative determination alanine aminotransferase [ALT] and albumin. Results revealed a non significant change in HCV-RNA and ALT in HCV-related HCC patients compared to HCV infected patients. Whereas, TGF-beta, TNF-alpha, AFP and NF-kB were increased significantly in HCV-related HCC patients compared to HCV infected patients. Stepwise multi-regression analysis showed that NF- kB [more than 1.49 fold change] with TGF-beta [more than 8438 pg/ml] together are the most sensitive predictors for HCC. NF- kB, TGF- beta are the most sensitive predictors for HCC on top of HCV

Sequence analysis of the hepatitis C virus non-structural 5A region [NS5A] in patients with hepatocellular carcinoma

The current study was aimed to analyze the sequence of hepatitis C virus non-structural 5A region [NS5A] from patients with hepatocellular carcinoma [HCC] on top of hepatitis C virus and compare these sequences with those of hepatitis C virus non-structural 5A region [NS5A] from patients with chronic active hepatitis C virus to characterize the similarity and/or differences between the two groups. This study included 20 male Egyptian patients, their age ranged from 43 to 74, with hepatitis C virus _ related hepatocellular carcinoma [HCV-related HCC] and no evidence of extrahepatic metastasis. All were negative for hepatitis B surface antigen [HBsAg]. All patients were HCV genotype-4a. Ten Egyptian patients with chronic active hepatitis C virus with matched age and sex were also included as reference group. Serum samples were collected for HCVRNA extraction, amplification and sequencing. Sequence analysis revealed that 11 patients out of 20 [55%] of the HCV-related HCC group harbored a wild-type strain sequence of NS5A region [Accession # NP

Risks and benefits of conservative management in severe pre-eclampsia

This prospective study was designed to detect whether conservative management in cases of severe pre-eclampsia is possible or not and to avoid prematurity and its related complications. This prospective case controlled study was conducted at Ain Shams University Maternity Hospital, over two years from April 2003 to April 2005, to detect whether conservative management in cases of severe pre-eclampsia is possible or not and to avoid prematurity and its related complications. Eighty five [85]: patients with severe pre-eclampsia [PET] were included in this prospective case controlled study. All patients included in this study were primigravida, not in labor, had a living fetus and had documented evidence of pre-eclampsia. The gestational age of the patients included in this study was more than 30 weeks gestation. Patients with maternal or fetal indications necessitating immediate delivery on admission were excluded from this study. All patients were admitted to the MICU of Ain Shams University Maternity Hospital. Before starting conservative management, patients and their relatives were informed about the management plan and possible maternal or fetal risks and benefits. If the patient agreed to continue conservative management, she signed a consent form. After counseling, forty five [45] patients refused the conservative management were included in the first group of this study [severe pre-eclampsia with immediate TOP], and conservative management was accepted by forty patients [40], included in the second of this study [severe pre-eclampsia managed conservatively]. The goal of the conservative management was to prolong pregnancy until 36 completed weeks [to avoid prematurity and to decrease the chance of neonatal incubation] or until the onset of either maternal or fetal complications [impending eclampsia or fetal distress]. The management was started immediately following admission with: bed rest, magnesium sulfate and an antihypertensive [methyldopa or nifedipine +/- hydralazine]. After control of blood pressure and prevention of convulsions, either TOP was started in the first group or conservative management in the second group. All patients in the second group received dexamethasone 12 mg IM every 12 hours for 48 hours and dexamethasone therapy was repeated weekly till delivery. 24 hours urinary total protein [UTP], Blood urea, creatinine, uric acid, SCOT, SGPT and platelet count were determined in all cases on admission. These laboratory investigations were repeated weekly for patients in the second group. Patients of the second group were monitored by senior obstetrician and senior anaesthiologist managing the MICU every day. Monitoring included inquiry about any complaints, blood pressure measurement 4 times daily, fetal heart sound auscultation 4 times daily, fetal growth monitoring, assessment of fetal wellbeing twice weekly and repetition of other tests whenever necessary. Patients were instructed to report the development of features such as persistent headaches, visual disturbances, epigastric pain, uterine contractions, vaginal bleeding, ruptured membranes or decreased fetal movement. TOP was based on both maternal and fetal indications. All patients were discharged after delivery with stable blood pressure levels and absent or + I urinary albumin. There was a high significant difference between the two groups of this study as regards the gestational age at TOP which was 32.4 weeks +/- 2.5 in the first group with immediate TOP compared with 35.4 weeks +/- 6.2 in the second group managed conservatively. Although, the first group with immediate TOP had shorter hospital stay before TOP [0.25 days +/- 0.5] than the second group managed conservatively [20.5 days +/- 4.5], the same group with immediate TOP had longer hospital stay after TOP [7.5 days +/- 3.5] titan the second group managed conservatively [5.0 days +/- 1.5]. The two groups of this study were matched with no significant difference as regards the mean systolic and diastolic blood pressure on admission, but the mean systolic blood pressure on TOP was significantly lower in the second group managed conservatively 140.8 mmHg +/- 12.5 compared with 155.5 mmHg +/- 22.5 in the first group with immediate TOP, also, the mean diastolic blood pressure on TOP was significantly lower in the second group managed conservatively 101.5 mmHg +/- 10.5 compared with 112.5 mmHg +/- 15.5 in the first group with immediate TOP. In the first group with immediate TOP, the serum uric acid was significantly decreased from 6.8 mg/dL +/- 1.73 on admission to 5.5 mg/dL +/- 0.35 on TOP, also, the SCOT was significantly decreased from 38.0 IU/L +/- 1.5 on admission to 31.2 IU/L +/- 3.58 on TOP and the SGPT was significantly decreased from 42.0 IU/L +/- 7.21 on admission to 20.0 IU/L +/- 5.7 on TOP. In the second group managed consewatively, the blood urea level was significantly decreased from 37.25 mg/dL +/- 7.32 on admission to 34.30 mg/dL +/- 5.67 on TOP, and the serum creatinine was significantly decreased from 1.35 mg/dL +/- 0.56 on admission to 1.24 mg/dL +/- 1.1 on TOP. Also, in the second group, the SCOT was significantly decreased from 40.50 IU/L +/- 3.5 on admission to 29.85 IU/L +/- 8.36 on TOP, and the SGPT was significantly decreased from 41.62 IU/L +/- 10.2 on admission to 19.8 IU/L +/- 7.35 on TOP, but there was a significant increase in the level of uric acid from 7.2 mg/dL +/- 1.12 on admission to 7.5 mg/dL +/- 0.87 on TOP and a significant decrease in the Platelet count from 192.907 x 1000/mm3 +/- 93.25 on admission to 156.484 x 1000/mm3 +/- 77.387 on TOP. The incidence of accidental hemorrhage was significantly higher in the first group of this study with immediate TOP [two patients = 0.44%], compared with the second group managed conservatively [one patient = 0.25%] and the incidence of antepartum eclampsia was signifantly higher in the second group managed consematively [3 patients = 0.75%] compared with the first group with immediate TOP [two patients - 0.44%]. In the second group of this study, The consewative management was successful and was continued till 36 weeks gestation in 22 patients out of 40 [55.0%]. 23 patients [57.5%] delivered vaginally in the second group managed conservatively compared with 21 patients [46.67%] in the first group with immediate TOP [P < 0.05 S.] and the number of the patients delivered by cesarean section was 24 out of 45 [53.33%] in the first group with immediate TOP and was 17 out of 40 [42.5%] in the second group managed conse-rvatively [P < 0.001, H.S.]. Also, there was a significant difference between the two groups of this study as regards fetal birth weight, which was 1.890 Kg +/- 0.850 in the first group with immediate TOP versus 2.425 Kg +/- 1.55 in the second group managed conservatively [P < 0.001, H.S.]. The number of the infants admitted to the NNICU was 29 out of 45 [64.4%] in the first group with immediate TOP compared with 12 out of 40 [30.0%] in the second group managed conservatively [P < 0.001, H.S.]. The neonatal deaths were significantly higher in the first group with immediate TOP [12 neonates - 26.7%] compared with the second group managed conservatively [3 neonates = 0.75%] [P < 0.001]. The duration of admission to NNICU was significantly longer in the first group with immediate TOP [37.5 days +/- 9.0] compared with the second group managed conservatively [19.0 days +/- 11.5] [P < 0.001]. The postpartum complications were significantly higher in the first group with immediate TOP compared with the second group managed conservatively. Two patients [0.44%] in the first group developed post-partum hemorrhage, while no cases of post-partum hemorrhage recorded in the second group [P < 0.001], also, one patient [0.22%] developed post-partum renal failure in the first group, while no cases of post-partum renal failure recorded in the second group [P < 0.05]. Six patients [13.4%] developed post-partum eclampsia in the first group compared with 3 patients [0.75%] in the second group [P < 0.001] and two patients [0.44%] developed HELLP syndrome in the first group compared with one patient [0.25%] in the second group [P < 0.05]. Consewative treatment with continuous monitoring and supervision is possible in selected cases of severe pre-eclampsia, in order to improve the fetal outcome in developing countries where NNICU is not available, without increasing the maternal risk

Prevalence of thyroid autoantibodies in Down syndrome

The prevalence of antithyroperoxidase antibodies in 40 infants and children with Down syndrome was determined and compared with their prevalence in 40 healthy children. The association of antithyroperoxidase antibodies and thyroid dysfunction in this group of Down syndrome patients was also studied. Twenty-six Down syndrome patients were positive for antithyroperoxidase antibodies; while in the control group, only 6 were positive. Thirteen Down syndrome patients had subclinical hypothyroidism, i.e. normal T4 and raised TSH [>5 ml U/ml]. Eight of them were positive for antithyroperoxidase antibodies. In the control group, five patients had sub-clinical hypothyroidism and two patients were positive for antithyroperoxidase antibodies. This supported the observation that the most frequently observed thyroid abnormality appears to be subclinical hypothyroidism in patients with Down syndrome

Plasma leptin, tumor necrosis factor-alpha and tissue inhibitor of metalloproteinase-1 in chronic hepatitis C patients and their relation to hepatic fibrosis

Hepatitis C virus [HCV] infection is a major cause of chronic hepatitis which frequently leads to progressive hepatic fibrosis and ultimately liver cirrhosis. Many humoral factors and cytokines are known to have profibrogenic and proinflammatory role in chronic hepatitis C patients [CHC] including leptin and TNF-alpha that act in an orchestrated manner with other factors to augment hepatic fibragenesis and. High circulating levels of leptin have been observed in CHC patients, and activated hepatic stellate cells [HSCS] are the effector cells producing leptin in addition to the adipose tissue cells. These HSCS secrete also tissue inhibitor of metalloproteinase-l [TIMP-1] which is one of TIMPS that interact with matrix metalloproteinases [MMPS] to regulate the turnover of extracellular matrix. The role of TIMP-1 in hepatic fibrogenesis and its possible value as a serum marker predicting liver fibrosis was reported in many studies. The aim of this study was to evaluate the possible role of leptin and TN F-alpha in hepatic. fibrogenesis and the possible role of TIMP-1 in determining the progression of liver fibrosis in a cohort of CHC patients. We aimed to evaluate the potential use of these serum markers to predict hepatic fibrosis. We measured leptin and TNF-alpha in the plasma of 55 CHC patients and 25 age and sex matched control persons by radioimmunoassay. Serum TIMP-1 was measured in both CHC patients and control persons by ELISA method. Liver disease in CHC patients was evaluated by the stage of hepatic fibrosis, grading of necroinflammation and grading of hepatic steatosis in liver biopsy. The mean age of CHC patients was 42.2 years [27-69 years] and the mean age of normal control persons was 4l.5[25-67 years]. Plasma - leptin, TNF-alpha and TIMP-1 were significantly higher in CHC patients compared to the normal controls [P<0.001 for leptin, P<0.05 for TNF-alpha, P<0.05 for TIMP-1]. Leptin correlated with BMI [body mass index] in CHC patients [males: r=0.51 P<0.01, [females: r=0.75 P<0.001], while both TNF-alpha and TIMP-1 did not correlate with any of clinical or biochemical parameters in CHC patients. Leptin levels increased as hepatic fibrosis stage progressed both in male and female CHC patients [males P<0.01, females P<0.001]. TNF-alpha levels and TIMP-1 levels were significantly higher in advanced stages of hepatic fibrosis [P<0.01 for TNF-alpha, P<0.001 forTIMP-1]. Plasma leptin and TNF-alpha levels showed no correlation with hepatic steatosis or histological necroinflammatory activity in CHC patients- Serum TIMP-1 levels did not correlate with hepatic steatosis but correlated with histological activity in CHC patients [Knodells score: r=0.91 P<0.001; Scheuer's Score: r=0.84 P<0.001]. Serum leptin levels did not correlate with TNF-a levels [r=0.18 P>0.05] or TIMP-1 levels [r=0.21 P>0.05]. Our data showed that serum leptin increased in CHC patients according to the severity of hepatic fibrosis. It seems to play a role in regulating hepatic fibrosis in addition to TNF-alpha and other profibrogenic cytokines. TIMP-1 levels increased similarly in advanced stage of hepatic fibrosis and it seems to be involved in determining the progression of hepatic fibrosis. These serum markers are potentially good tools to assess hepatic fibrosis in CHC patients and they may be useful to assess the response to antiviral therapy. They may represent therapeutic targets in the future to modulate or stop hepatic fibrosis

Relevance of recombinant immunoblot assay III to HCV-RNA polymerase chain reaction positivity and IGM antibody to HCV core antigen for HCV infection confirmation

Currently the second and third generation enzyme immunoassays [EIA2-EIA3] for hepatitis Virus antibody [anti-HCV] are the most practical screening tests for the diagnosis of HCV infection. The need for confirmatory test depends on the clinical setting and the likelihood of a true - positive EIA result. The most appropriate approach is to retest for anti-HCV using recombinant immuno-blot assay [RIBA] and then test for HCV-RNA using PCR assay in those who are RIBA positive or indeterminate. A third generation supplemental test [RIBA III] has been introduced. It appears to be more specific test based on a better correlation with RNA PCR results with a reduced number of indeterminate results. To study the clinical Value of the recombinant immunoblot assay [RIBA] in routine diagnostics of patients with HCV infection using HCV PCR as the most definitive test We further studied the correlation between HCV core IgM positivity and HCV RNA detection by PCR. Patients: 47 patients with chronic hepatitis after more than 6 months disease duration. Outcome measures: Liver function tests [ALT, AST, GGT], Hepatitis C virus antibody IgG by Enzyme immunoassay version III; HCV antibody IgM by ELISA technique, HCV antibody using Recombinant Immunobloting Assay [RIBA]; quantitative HCV RNA estimation using amplisensor assay after RNA extraction. high viremia was considered in samples with more than 106 copies/ml and low viremia with samples less than 10[6] copies/ml. Negative reaction was considered when no reaction was obtained. Significantly greater number of patients with HbcIgG+ve/HCV IgM +ve had +ve NS5 b and when compared to those with HBc IgG -ve/HCV IgM+ve. Those with HBc IgG +ve/HCV IgM-ve showed highly significantly fewer number of positive C3+C4 b and cases when compared to the HBcIgG ve/HCV IgM+ve and HbcIgG+ve/HCV IgM +ve patients. Likewise, highly significant fewer number of positive C1+C2 cases with HBc IgG +ve/HCV IgM-ve were obtained when compared to cases with HBcIgG -ve/HCV IgM+ve and HbcIgG+ve/HCV IgM +ve patients. our findings suggest that ELISA-3 was at par with RIBA III and it can be recommended for routine screening for anti-HCV. However, despite the cost, the combination of both ELISA-3 and qualitative or quantitative PCR for HCV RNA allows a definitive classification of HCV diagnosis

Urine neopterin: a new parameter for monitoring disease activity in patients with systemic lupus erythematosus

Urine neopterin/creatinine [N/C] ratio was determined in early morning samples of 16 children with active systemic lupus erythematosus [SLE] and 10 children with inactive SLE disease, using radioimmunoassay [RIA] for measuring urine neopterin Concentration. Compared with apparently healthy 20 control children the mean urine N/C ratio was significantly increased in patients with active [P <0.001] and inactive [P <0.01] SLE. The mean urine N/C ratio was also significantly higher in children with active than in. those with inactive SLE [P <0.001]. The urine N/C ratio did not distinguish between patients with renal activity and those with other types of clinical activity and it was not influenced by different drug regimens. The correlation between urine N/C ratio and erythrocyte sedimentation rate [ESR] was a positive linear one while there was no significant correlation between urine N/C ratio and serum C4. When testing urine N/C ratio at a cut-off value of >/= 300 micro mol neopterin/mol creatinine and ESR at a cut-off value of >/= 50 mm/hr and serum C4 at a cut-off value of