The accuracy and cost-effectiveness of hepatitis C core antigen assay in the monitoring of anti-viral therapy in patients with chronic hepatitis C genotype 4.

BACKGROUND: Quantitative hepatitis C (HCV) polymerase chain reaction (qtHCV-PCR), the gold standard for monitoring HCV therapy, is an expensive, time-consuming procedure that requires equipped laboratories and trained personnel. AIMS: To assess the accuracy and cost-effectiveness of the automated Abbott ARCHITECT HCV Ag assay (Wiesbaden, Germany) in monitoring response to pagylated interferon (PEG-IFN) and ribavirin therapy for chronic HCV genotype 4 (G4). METHODS: This longitudinal, non-inferiority study compared the efficacy and cost benefit of an All-HCV core antigen assay protocol and a hybrid qtHCV RNA PCR and HCV core Ag assay protocol to the standard All-qtHCV-PCR protocol in chronic HCV G4 patients treated with pegylated interferon (PEG IFN) and ribavirin. RESULTS: Four hundred and ten patients with chronic hepatitis C genotype 4 met inclusion criteria and were enrolled in the study. The sustained virological response rate (SVR) was 66.34%. The All-HCV core antigen and hybrid monitoring assays resulted in the significant cost savings without compromising performance. A good correlation existed between HCV viral load and HCV core antigen levels (r = 0.944; P < 0.0001). Baseline HCV RNA values <600 IU/mL, baseline HCV core Ag levels <2000 fmol/L, rapid virological response, rapid decline in HCV core antigen were strong predictors of SVR. CONCLUSION: Hepatitis C core Ag assay is a point-of-care, reproducible, reliable, cost-effective monitoring tool with rapid turnaround time that, which can effectively replace or adjunct to qRT-PCR in monitoring interferon based or interferon-free anti-viral therapy in chronic hepatitis genotype 4 patients in resource-limited countries with a high burden of hepatitis C.

Near-infrared spectroscopic assessment of mitochondrial oxygenation status--comparison during normothermic extracorporeal liver perfusion by buffer only or buffer fortified with washed red blood cells: an experimental study.

BACKGROUND: Use of marginal and non-heart-beating donors leads to an increased incidence of complications after clinical liver transplantation. Normothermic extracorporeal liver perfusion (NELP) may allow resuscitation and evaluation of such organs. Despite recent success in long-term liver preservation by NELP, no methods of organ evaluation have been defined. Mitochondrial cytochrome oxidase (Cyt Ox) levels reflect oxygen and substrate delivery, and hence ATP production at the cellular level. This study used near-infrared spectroscopy (NIRS) to measure Cyt Ox levels during NELP. METHODS: Livers retrieved from New Zealand white rabbits were immediately perfused in an extracorporeal circuit with oxygenated buffer (group A, n = 4) or red blood cell (RBC)-fortified buffer (group B, n = 4). Perfusion was continued for 3 hours at 37 degrees C pH 7.4, and perfusate was gassed with 95%O2/5%CO2 at 1 liter per minute. Cyt Ox levels were monitored continuously by NIRS and bile output was measured. RESULTS: Cyt Ox was reduced at the start of perfusion in both groups, but even more rapidly in the buffer-perfused group. After initial deterioration, Cyt Ox levels improved significantly (P < .05) with perfusion in the RBG-perfused group, but remained impaired in the buffer group 5.74 +/- 1.51 Deltamicromol/L and -25.77 +/- 21.94 Deltamicromol/L for groups B and A, respectively, at 180 minutes. Differences in bile output were not significant (19.33 +/- 9.50 and 25.00 +/- 16.81 micromol/min/100 g for groups B and A respectively). CONCLUSIONS: Cyt Ox levels may offer better viability markers than bile output. NIRS is a practical method to measure tissue oxygenation, and RBC-based perfusion provided better oxygenation during NELP.

A comparison of bile composition from heart-beating and non-heart-beating rabbit organ donors during normothermic extracorporeal liver perfusion: experimental evaluation using proton magnetic resonance spectroscopy.

BACKGROUND: Organs retrieved from marginal and non-heart-beating donors (NHBDs) have sustained variable degrees of preretrieval damage that result in an increased incidence of complications. Normothermic extracorporeal liver perfusion (NELP) provides an opportunity to evaluate and resuscitate such organs. The aim of this study was to identify markers of ischemic injury in bile during perfusion of livers from heart-beating donors (HBDs) and NHBDs. METHODS: Livers were retrieved from New Zealand white rabbits. The HBD group (n=4) had no in situ warm ischemia before retrieval and the NHBD group (n=4), 45 minutes of in situ warm ischemia before liver retrieval. After 40 minutes of postretrieval cold ischemia, all livers were dual vessel reperfused, normothermically with oxygenated buffer solution supplemented with rabbit red blood cells, for 6 hours. Bile was collected and examined with 1HMRS. RESULTS: Perfusion bile from HBD group showed an increased concentration of bile acids, lactate, glucose, and phosphatidylcholine, but a decreased concentration of acetate as compared to retrieval bile. This trend was further enhanced in NHBD group. The mean +/- SD (in micromol/L) were bile acids (10.48 +/- 2.8 vs 26.05 +/- 12.1 vs 44.5 +/- 44.5), lactate (10.66 +/- 4.5 vs 14.66 +/- 5.2 vs 13.22 +/- 1.8), glucose (5.37 +/- 2 vs 21.2 +/- 5.0 vs 29.09 +/- 15.3), phosphatidylcholine (0.21 +/- 0.02 vs 5.57 +/- 1.7 vs 6.42 +/- 0.3), and acetate (1.8 +/- 0.5 vs 0.39 +/- 0.1 vs 0.38 +/- 0.09) for retrieval bile, HBD perfusion bile, and NHBD perfusion bile, respectively. One animal from each group did not produce any bile during perfusion. CONCLUSIONS: 1HMRS of biliary constituents revealed differences with the type of ischemia. These indices may be potential markers of the extent of warm ischemic injury and the functional activity of an extracorporeally perfused liver.

The possible meaning of transferrin and its soluble receptors in seminal plasma as markers of the seminiferous epithelium.

Transferrin (Tf) and soluble transferrin receptors (S-Tf-R) were measured by enzyme immunoassay in seminal plasma of 130 semen samples. The mean concentration of S-Tf-R in cases with normozoospermia was 10.4 IU/ml (95% confidence interval: 9.5-11.3) and it was significantly lower in patients with oligozoospermia (6.6, 95% CI: 5.8-7.5, P < 0.001), asthenozoospermia (8.5, 95% CI: 5.5-10.7, P < 0.05), azoospermia of primary testicular origin (7.9, 95% CI: 6.1-9.6, P<0.05) and post-vasectomy samples (5.9, 95% CI: 5.4-6.9, P < 0.001). The concentration of S-Tf-R in post-vasectomy samples was lower than that in patients with azoospermia of primary testicular origin (P < 0.05; positive likelihood ration = 7 at value of 8.3 IU/ ml). S-Tf-R was positively correlated with motile sperm concentration (r = 0.50, P < 0.0001), percentage motility (r = 0.38, P < 0.001), percentage of normal forms (r = 0.43, P < 0.001), sperm linear velocity (r = 0.42, P < 0.001), and ATP concentration (r = 0.67, P < 0.0001). Follicle stimulating hormone (FSH) was found to be negatively correlated with the concentrations of both Tf (r = -0.31, P < 0.05) and of S-Tf-R (r = -0.45, P < 0.01). The mean concentration of Tf in seminal plasma was 50.4 micrograms/ml (35.9-67.2) in samples with normozoospermia (n = 22), and the concentration was significantly lower in patients with oligozoospermia (P < 0.05), azoospermia of testicular origin (P < 0.001), and post-vasectomy samples (P < 0.001). Seminal Tf was correlated with motile sperm concentration (r = 0.36, P < 0.001), percentage of motile spermatozoa (r = 0.25, P < 0.05), linear velocity (r = 0.24, P < 0.05) and ATP concentration (r = p.44, P < 0.001). The concentration of Tf was positively correlated with that of S-Tf-R both in cases with spermatozoa present (r = 0.66, P < 0.001), and in cases with azoospermia of testicular origin (r = 0.51, P < 0.05) but not in vasectomy cases. It is concluded that S-Tf-R in seminal plasma is a marker of spermatogenesis and may give information on the presence or absence of spermatogenetic cells in cases with azoospermia. Further investigations are needed to assess its usefulness for clinical practice.

Evaluation of beta-endorphin and interleukin-6 in seminal plasma of patients with certain andrological diseases.

Human semen contains large amounts of opioid peptides and cytokines. We have measured the concentrations of interleukin (IL)-6 in 140 semen samples and of beta-endorphin in 77 semen samples. The median concentration of beta-endorphin in seminal plasma from normozoospermic men (n = 23) was 154.7 pg/ml (10th-90th percentiles, 42.0-774.6), and there was no significant difference in the beta-endorphin concentration among normozoospermic, oligozoospermic (n = 28), asthenozoospermic (n = 15), azoospermic (n = 4) and post-vasectomy (n = 7) samples. There was no correlation between beta-endorphin concentration and sperm characteristics, nor with blood hormones. beta-Endorphin concentration was lower in cases with immunological infertility, as revealed by a positive direct mixed antiglobulin reaction test (n = 12) (P < 0.01), than in matched controls. The median concentration of IL-6 in samples with normal sperm concentration, motility and morphology with or without white blood cells (n = 39) was 26.1 pg/ml (10th-90th percentiles, 7.3-172.3), and there was no significant difference in the IL-6 concentration among normozoospermic, oligozoospermic (n = 46), asthenozoospermic (n = 32), azoospermic (n = 13) and post-vasectomy (n = 10) samples. The IL-6 concentration was significantly higher in cases of varicocele (n = 22) without white blood cells in semen (P < 0.001) than in matched controls without varicocele (n = 23). In addition, the IL-6 concentration was elevated (P < 0.0001) in cases with accessory sex gland inflammation (n = 40). IL-6 concentration was positively correlated with white blood cells in semen (n = 60, r = 0.59, P < 0.0001), but there was no correlation with beta-endorphin concentration. The IL-6 concentration chosen to differentiate between cases with and without accessory gland inflammation was 45.3 pg/ml, with a specificity of 80.6% and a sensitivity of 92.5%. It is concluded that beta-endorphin in seminal plasma plays an immune suppressive role, and that increased IL-6 concentration may be related to testicular dysfunction in cases with varicocele. Furthermore, IL-6 is an accurate marker of accessory sex gland inflammation.

Androgens in seminal plasma: markers of the surface epithelium of the male reproductive tract.

Androgens in seminal plasma have been investigated in relation to other markers of the male reproductive system. Testosterone and dihydrotestosterone (DHT) were measured by radioimmunoassay after paper chromatography in 104 semen samples. Concentrations of testosterone and DHT were significantly lower in samples from patients with abnormal sperm characteristics than in men with normozoospermia. Both testosterone and DHT were correlated significantly with sperm concentration (r = 0.40, p < 0.001; r = 0.41, p < 0.001, respectively), motile sperm concentration (r = 0.35, p < 0.01; r = 0.35, p < 0.01) and ATP concentration (r = 0.59, p < 0.001; r = 0.45, p < 0.001). In addition, the total amounts of testosterone and DHT were correlated with total activity of alpha-glucosidase (r = 0.49, p < 0.001; r = 0.58, p < 0.01), and gamma-glutamyltransferase (r = 0.49, p < 0.001; r = 0.48, p < 0.001) in seminal plasma. Transferrin (Tf) concentration in seminal plasma was significantly lower in samples without spermatozoa, and total Tf content was lower in oligozoospermic samples (p < 0.05). Testosterone and DHT were correlated positively with Tf levels in seminal plasma (r = 0.48, p < 0.0001; r = 0.78, p < 0.0001 respectively). Testosterone, DHT and Tf in seminal plasma were higher in the first than in the second fraction of split ejaculates, and DHT, but not testosterone, increased significantly in each of six patients treated with tamoxifen. Testosterone, DHT, and Tf in seminal plasma were not correlated with serum concentrations of LH, FSH, testosterone or prolactin, but the DHT : T ratio in seminal plasma was correlated with serum LH (r = 0.36, p < 0.05). It is concluded that the concentration of androgens in seminal plasma is related predominantly to the activity of the seminiferous epithelium and, to a lesser extent, to the function of the accessory sex glands.

Evaluation of the role of reactive oxygen species in male infertility.

Reactive oxygen species (ROS) production from spermatozoa has been measured by chemiluminescence in the two fractions of a Percoll gradient column (47 and 90%). Chemiluminescent signals were recorded in each fraction after the addition of luminol and horse-radish peroxidase (basal state), and after stimulation with formyl-methionyl-leucyl-phenylalanine and phorbol ester (PMA). Oligozoospermic samples show a higher rate of ROS production than the normozoospermic samples in both fractions of Percoll. Also, ROS were generated at a higher rate by asthenozoospermic samples in the 90% Percoll fraction than by normal samples after stimulation with PMA. Our data confirm that fact that white blood cells play a major role in the production of ROS, even after purification on a Percoll gradient. Immunological cases were also found to be associated with an increased production of ROS, which may be caused by the same underlying pathological condition responsible for the production of the antibodies. Repeated centrifugation of the samples triggers a burst of ROS in excess of that produced after Percoll preparation. In addition, superoxide dismutase activity was found to be significantly increased in cases with an elevated production of ROS. It is concluded that measuring the ROS generation by semen may yield useful information on the functional capacity of spermatozoa, which may be used to improve the success of male infertility management.

Relationship between resazurin reduction test, reactive oxygen species generation, and gamma-glutamyltransferase.

The production of reactive oxygen species (ROS) from spermatozoa has been measured by chemiluminescence in the two fractions of a Percoll gradient column (47 and 90%). Chemiluminescent signals were recorded in each fraction after addition of luminol and horse-radish peroxidase (basal state), and after stimulation with formyl-methionyl-leucyl-phenyl-alanine (FMLP) and phorbol ester (PMA). We found an inverse correlation between the reducing capacity of the semen as estimated by the resazurin test, and the production of ROS by the spermatozoa, especially after stimulation with PMA (r = 0.51, P < 0.001). gamma-glutamyltransferase activity of seminal plasma was inversely correlated with ROS (r = -0.47, P < 0.01) and the resazurin test result (r = -0.43, P < 0.001) suggesting a possible role of prostatic secretions in the protection of spermatozoa against oxygen radicals. The resazurin test has a positive predictive value of 92.5% at a criterion value of colour scale 4 to discriminate between normal and excessive ROS production by spermatozoa, with sensitivity 79% and specificity 74%. In addition, at criterion value of colour scale 3, the resazurin test could distinguish between samples with normal or low activity of gamma-glutamyltransferase, with sensitivity 57% and specificity 93%. It is concluded that the result of the resazurin test can be influenced by the oxidative stress of spermatozoa and by prostatic function.

Serum mineral changes in alloxan diabetes before and after treatment with some hypoglycemic drugs.

In alloxan diabetes, serum zinc, copper, iron and magnesium were significantly higher than in normal rats, while the level of serum calcium, sodium, and potassium was lower than normal. Treatment with daonil or insulin led to a normalization, as expected of the level of serum glucose and most of the other elements, except for iron and potassium. When lycanol was used for treatment, the level of all elements returned to the normal except for blood glucose, zinc and potassium.

Clinical, biochemical and histochemical studies on infants with Acrodermatitis enteropathica chronica.

7 infants diseased with Acrodermatitis enteropathica and 10 normal controls were included in this study. The values of anthranilic acid glucuronide, 6- aminohippuric, anthranilic acid, N-acetyl Kneurine, Kneurine and 30 H Kneurenine, were estimated in mg/24 hours urine, both basal and after tryptophane load. In addition, histopathological and histochemical studies for lactase, succinic dehydrogenase, alkaline phosphatase, acid phosphatase, and alpha-non-specific esterases activities were done for the intestinal mucosal biopsies. All the previous investigations were then repeated after two months treatment with 500 mg/day diiodohydroxyquinoline. The tryptophan metabolites were significantly low in the diseased infants, both basal and after tryptophan load. Moreover, the intestinal enzymes activities were altered. After 2 months treatment with diiodohydroxyquinoline the diseased infants became clinically improved, tryphtophan metabolites became normal, but the activities of the intestinal enzymes were not altered. The biochemical and histochemical findings were discussed, giving the possibility of competitive inhibition of the diiodohydroxyquinolines and the by-product 8 OH Quinololic acid resulting in more degradation of Kneurine and 3 OH Kneurenine to nicotinamide adenine dinucleotide.

Calcium and phosphorus metabolic changes in children with hepatic bilharziasis.

Investigations were done to study the calcium and phosphorus metabolic changes in 35 children with hepatic bilharziasis, of whom 12 were ascitic. Thirteen normal children served as controls. Despite a normal serum level of both calcium and inorganic phosphate in the Bilharzial group, yet both were increased in urine. The mean values obtained from the ascitic fluid were 70% and 90.2% of their serum levels, respectively. The corrected creatinine clearance was significantly diminished, while PEI was significantly increased, and the % TRP significantly decreased, with still a more marked decrease in the ascitic cases. Following a phosphate lead the serum inorganic phosphate level showed a significantly more rapid lowering in both bilharzial groups, as compared to the control group. On the other hand bilharzial cases responded to the intravenous calcium load by a significant retention of calcium in the serum.