Further observations on hydrogen peroxide antisepsis and COVID-19 cases among healthcare workers and inpatients.

BACKGROUND: The use of prophylactic antisepsis to protect against coronavirus disease 2019 (COVID-19) has been suggested. This study investigated hydrogen peroxide antisepsis (HPA) at two hospitals in Ghana. METHODS: Cases of COVID-19 among healthcare workers (HCWs) using hydrogen peroxide (HP-HCWs) or not using hydrogen peroxide (NHP-HCWs), vaccinated or unvaccinated, were recorded at Shai-Osudoku Hospital (SODH), Dodowa, and Mount Olives Hospital (MOH), Techiman, between May 2020 and December 2021. The effect of HPA in all inpatients at MOH was also observed. Permutation tests were used to determine P values. FINDINGS: At SODH, there were 62 (13.5%) cases of COVID-19 among 458 NHP-HCWs but no cases among eight HP-HCWs (P=0.622) from May to December 2020. Between January and March 2021, 10 (2.7%) of 372 NHP-HCWs had COVID-19, but there were no cases among 94 HP-HCWs (P=0.206). At MOH, prior to HPA, 17 (20.2%) of 84 HCWs and five (1.4%) of 370 inpatients had COVID-19 in July 2020. From August 2020 to March 2021, two of 54 (3.7%) HCWs who stopped HPA had COVID-19; none of 32 NHP-HCWs contracted COVID-19. At SODH, none of 23 unvaccinated HP-HCWs and 35 (64%) of 55 unvaccinated NHP-HCWs had COVID-19 from April to December 2021 (P<0.0001). None of 34 vaccinated HP-HCWs and 53 (13.6%) of 390 vaccinated NHP-HCWs had COVID-19 (P=0.015). No inpatients on prophylactic HPA (total 7736) contracted COVID-19. CONCLUSION: Regular, daily HPA protects HCWs from COVID-19, and curtails nosocomial spread of SARS-CoV-2.

Typing of human cytomegalovirus clinical isolates from Saudi patients by PCR-RFLP.

BACKGROUND: Information on strain types of human cytomegalovirus (HCMV) isolates from Saudi Arabian patients is lacking. MATERIALS AND METHODS: 80 clinical isolates of HCMV from Saudi Arabian patients were analyzed by PCR amplification of three regions (DNA polymerase, glycoprotein B, and glycoprotein H) of the virus genome. The resultant amplicons (2.0-2.7 kb) were further studied by restriction fragment length polymorphism (RFLP) using four enzymes (HaeIII, HhaI, MspI, and RsaI). RESULTS: Combined analysis of the cleavage patterns generated by the enzymes identified five strains, S1-S5, and several mixed and unique strains. 18 isolates belonged to S1 strain and were similar to laboratory strain AD169. Eight isolates were present in each of S2 and S3 strains. Six isolates and four isolates were found in S4 and S5 strains, respectively. 12 isolates contained a mixture of S3 and S5, which may have resulted from a dual infection. Each of the 24 remaining isolates had a different strain pattern. CONCLUSION: Our findings show that 80 HCMV clinical isolates were distributed into 30 different strains using PCRRFLP analysis of multiple viral subgenomic regions. However, the number of isolates is not uniformly distributed among strains (p < 0.02).

Peginterferon alpha-2b plus ribavirin compared with interferon alpha-2b plus ribavirin for initial treatment of chronic hepatitis C in Saudi patients commonly infected with genotype 4.

AIM: Comparing the efficacy of peginterferon alpha-2b plus ribavirin with interferon alpha -2b plus ribavirin in Saudi patients with chronic hepatitis C virus (HCV) commonly infected with genotype 4. METHODS: A total of 96 patients with chronic HCV infection were randomly assigned to two treatment groups. Forty-eight patients received once weekly 100 microg of peginterferon alpha-2b plus ribavirin given orally 800 mg/day (peginterferon group). Another 48 patients received thrice weekly 3 million units of interferon alpha-2b plus ribavirin 800 mg/day (interferon group). At the end of treatment (48 weeks) and sustained (72 weeks) biochemical and virologic responses were determined. RESULTS: In the peginterferon group, 70.8% (34/48) patients attained both biochemical and virologic responses at the end of the treatment as against 52.1% (25/48) patients in the interferon group. (P=0.09 for both). Similarly, sustained biochemical and virologic responses in the peginterferon group were attained in 52.1% (25/48) and 43.8% (21/48) patients as against 43.8% (21/48) and 29.2% (14/48) patients in the interferon group, respectively (P=0.54 and 0.20, respectively). The sustained virologic response rates in patients with genotype 4 were 42.9% (12/28) in the peginterferon group and 32.3% (10/31) in the interferon group (P=0.43). Patients in peginterferon group had higher, although statistically not significant adverse reactions. CONCLUSIONS: Saudi patients with chronic HCV attained a higher, although statistically not significant sustained virologic response with pegylated interferon plus ribavirin compared with interferon plus ribavirin.

Hand contamination with hepatitis C virus in staff looking after hepatitis C-positive hemodialysis patients.

BACKGROUND: Hepatitis C virus (HCV) is a major cause of hepatitis in hemodialysis (HD) patients. Routes other than blood transfusion play a role in the spread of HCV in HD patients. Molecular studies of HCV implicate nosocomial transmission of the virus in HD units. We conducted a clinicovirological study in our HD unit to investigate if the hands of dialysis personnel could represent a mode of transmission of HCV among HD patients. METHODS: One liter of sterile water was used for each handwashing of dialysis personnel. The washing was collected in a sterile container and tested for HCV-RNA by polymerase chain reaction (PCR) within 3 h of collection. Eighty handwashings from nurses dialyzing HCV-positive patients (groupe A) and 100 handwashing from nurses dialyzing HCV-negative patients (group B) were tested for HCV-RNA. As a control, 60 handwashings were collected from the dialysis personnel before entering the dialysis unit (group C) and tested for HCV-RNA. RESULTS: HCV-RNA was positive in 19 (23.75%) of samples of group A, in 8 (8%) of samples of group B (p < 0.003) and in 2 (3.3%) of samples of group C (p < 0. 35). These two positive samples of group C were from nurses who had dialyzed HCV-negative patients. CONCLUSION: These results indicate the presence of HCV-RNA on the hands of some dialysis personnel in our HD unit, in spite fo adherence to the standard precautions. The hands of dialysis personnel are therefore a potential mode for facilitating transmission of HCV between HD patients.

GB virus C/hepatitis G virus infection in Saudi Arabian blood donors and patients with cryptogenic hepatitis.

Viral hepatitis is a common infection in the developing countries. Aside from Hepatitis A-E viruses, a novel hepatitis virus termed GBV-C, or HGV, was recently described. We have studied the prevalence of this virus among Saudi Arabian healthy blood donors (n = 200) and patients with cryptogenic (non-A-E) hepatitis (n=71). After serum extraction and RNA reverse transcription, amplification was carried out by the polymerase chain reaction (PCR), using primers for the 5' noncoding region (NCR), NS5A region and NS3 helicase region. Among the patients with cryptogenic hepatitis, PCR-positivity was 18/71 (25.4%) for the 5' NCR, 14/71 (19.7%) for the NS5A region, and 15/71 (21.1%) for the NS3 helicase region. Among the healthy blood donors, PCR-positivity was 4/200 (2%) for the 5' NCR, 0/200 (0%) for the NS5A region, and 1/200 (0.5%) for the NS3 helicase region. Since the 5' NCR is considered the most conserved segment of the virus genome, it is not unusual to find higher positivity rate when that region is used for amplification. It is noted that the positivity rate is not far different among the three amplified regions, indicating that the heterogeneity of GBV-C/HGV is not as extensive as in hepatitis C virus. Phylogenetic analysis of 5'NCR DNA sequences showed that all isolates in this study belong to genotype 2. We conclude that the prevalence of GBV-C/HGV is similar to what is reported worldwide among the general Saudi population but relatively higher among Saudi patients with cryptogenic hepatitis.

Plasmid profile and antibiotic resistance in coagulase-negative staphylococci isolated from polluted water.

Four different species of coagulase-negative staphylococci (CNS) were isolated from polluted waters in Fez, Morocco and found to be Staphylococcus simulans, Staph. lenticus, Staph. hyicus and Staph. xylosus. Eight isolates belonging to these four species were analysed for their plasmid content. Southern blot hybridizations were performed to define the resistance determinants of the plasmids harboured by these species. These determinants were found to be carried mainly by Class I staphylococcal plasmids (1-5 kb). A plasmid (4.3 kb) carrying a tetracycline resistance gene was present in five isolates from all identified species. Plasmids carrying a chloramphenicol resistance gene were more frequently encountered and found to be of different sizes. Plasmids carrying erythromycin, neomycin, and streptomycin resistance genes were less frequent and were the same size. The results indicate that the occurrence of multi-resistant CNS in polluted waters may constitute a reservoir for disseminating antibiotic-resistance into the community.

The alpha chemokine, interleukin 8, inhibits the antiviral action of interferon alpha.

Interferon (IFN) exhibits a potent antiviral activity in vitro and plays a major role in the early defense against viruses. Like IFN, the proinflammatory chemokine, interleukin (IL)-8, is induced by viruses and appears in circulation during viral infections. In an in vitro cytopathic effect assay for IFN, we found that IL-8 can inhibit IFN-alpha activity in a dose-dependent manner. This action was reversed by specific monoclonal antibodies to IL-8. The chemokine was able to attenuate the IFN-mediated inhibition of viral replication as determined by measuring infectious virus yield. IL-8 also diminished the ability of IFN to inhibit an early stage of viral replication since IL-8 attenuated the inhibition of the formation of viral proteins. It appeared that IL-8 interfered with a late rather than an early step of IFN-mediated pathway such as early gene expression. The IL-8 inhibitory action on IFN-alpha antiviral activity was associated with reduced 2',5'-A oligoadenylate synthetase activity, a pathway well correlative with the anti- encephalomyocarditis virus action of IFN-alpha. Understanding pathways that antagonize IFN action may lead to novel approaches to potentiate endogenous and therapeutic IFN.

Serological diagnosis of hepatitis C virus in patients with liver disease in Saudi Arabia. Evaluation of antibody determination by recombinant immunoblot assays in relation to RNA detection by polymerase chain reaction.

Sera from 164 Saudi Arabian patients with non-A, non-B hepatitis liver disease were examined for antibodies to hepatitis C virus (HCV) by second- and third-generation recombinant immunoblot assay (RIBA-2 and RIBA-3) and for HCV RNA by polymerase chain reaction (PCR). By using RIBA-2, 92 (56.1%) were reactive, 64 (39%) were nonreactive, and 8 (4.9%) were indeterminate. By using RIBA-3, 98 (59.7%) were reactive 60 (36.6%) were nonreactive, and 6 (3.7%) were indeterminate. By using PCR, 108 (65.9%) were positive. Of the eight RIBA-2 indeterminate samples, seven became RIBA-3 reactive but PCR-positive, and one became RIBA-3 nonreactive but PCR-negative. Of the six RIBA-3 indeterminate samples, five were RIBA-2 nonreactive but PCR-positive, and one was RIBA-2 reactive but PCR-negative. From our study on Saudi patients, we conclude that RIBA-3 has slightly but not significantly improved the results of anti-HCV antibody detection, and is probably of more value to resolve those indeterminate samples by RIBA-2. Although expensive, PCR remains the most reliable HCV diagnostic method until an HCV antigen detection test is available.

Genetic homogeneity among methicillin-resistant Staphylococcus aureus strains from Saudi Arabia.

Ninety-four strains of methicillin-resistant Staphylococcus aureus (MRSA) were collected from patients nursed in several hospitals in Saudi Arabia, before they were referred to King Faisal Specialist Hospital and Research Centre for tertiary care. The hospitals were from geographically diverse regions and as such the entirety of Saudi Arabia was covered. All strains were genetically typed by random amplification of polymorphic DNA (RAPD) analysis using three different primers and a representative subset of the strains was analyzed with pulsed-field gel electrophoresis (PFGE) as well. It was concluded that 87 out of 94 (93%) belong to a single clonally related lineage of MRSA. In the other 7 cases, the DNA banding patterns were shown to differ only slightly from those determined for the clonal type. PFGE analysis confirmed the homogeneity of the collection of strains. When the RAPD and PFGE fingerprints obtained for the Saudi clone were compared to those generated for a collection of MRSA with a more diverse geographical background, it was shown that the clonal type from Saudi Arabia was not identical to any of these MRSA strains. Our data provide another example of the capacity of certain MRSA clones to expand through entire nations and establish themselves permanently among large number of hospitals and, consequently, even larger numbers of patients.

Genotyping of hepatitis C virus isolates from Saudi patients by analysis of sequences from PCR-amplified core region of the virus genome.

We investigated the genotype distribution of hepatitis C virus (HCV) among Saudi patients with chronic hepatitis C. Serum specimens from 119 native Saudi Arabian patients with chronic hepatitis C, as documented by serology and polymerase chain reaction (PCR) for HCV RNA, were used. Genotyping was performed by reverse transcription-PCR, using specific primers at the core region of HCV genome, and DNA sequencing of the resultant amplicons. It was found that the majority of samples (47.9%) belong to genotype 4, followed by subtype 1b (16.8%), and subtype 1a (10.1%). Twenty samples (16.8%) were not able to be typed by our method. We confirmed the results by cloning at least one PCR amplicon from each genotype, and determining the nucleotide sequence of the clones. Our findings suggest that genotype 4 is the most common among native Saudi Arabian patients with chronic hepatitis C infection. Genotypes 1b and 1a were also prevalent.

Hepatitis C virus infection in hemodialysis patients: comparison of two new hepatitis C antibody assays with a second-generation assay.

The performance of two new hepatitis C virus antibody (anti-HCV) assays (a third-generation immunoglobulin (Ig)G recombinant immunoblot assay (RIBA 3.0) and hepatitis C virus core IgM (HCV IgM) in the prediction of hepatitis C viremia in hemodialysis patients was compared with that of a second-generation IgG recombinant immunoblot assay (RIBA 2.0). Forty-three patients on maintenance hemodialysis were studied. Aliquots of sera were tested prospectively for anti-HCV by RIBA 2.0, RIBA 3.0, and HCV IgM and for HCV RNA by polymerase chain reaction. Thirty-eight patients were HCV RNA positive. Among those, 7 (18%) were HCV IgM positive, 22 (58%) were RIBA 2.0 positive, and 29 (76%) were RIBA 3.0 positive. All but one viremic patients detected by HCV IgM were also detected by RIBA 2.0 and RIBA 3.0. All viremic patients detected by RIBA 2.0 were also detected by RIBA 3.0. RIBA 3.0 was more sensitive than RIBA 2.0 and HCV IgM in the detection of viremic patients (P = 0.0156 and < 0.0001, respectively). The positive predictive value for HCV IgM was 100% as compared with 96 and 97% for RIBA 2.0 and RIBA 3.0, respectively. The negative predictive value for RIBA 3.0 was 36% as compared with 24 and 14% for RIBA 2.0 and HCV IgM, respectively. At 6-months follow-up of the eight viremic patients without a detectable IgM or IgG anti-HCV response, all patients remained RIBA 2.0 nonreactive, one became RIBA 3.0 indeterminate, and one became HCV IgM positive. These data suggest that HCV IgM has poor sensitivity in the detection of hepatitis C viremia and RIBA 3.0 improves the sensitivity of IgG anti-HCV assays in the early detection of hepatitis C viremia in hemodialysis patients.

New insights into hepatitis C virus infection of hemodialysis patients: the implications.

The authors compared the diagnostic performance of a second-generation recombinant immunoblot assay (RIBA) (RIBA HCV 2.0 SIA) and the recently introduced third-generation RIBA (RIBA HCV 3.0 SIA) with that of hepatitis C virus (HCV) RNA by the polymerase chain reaction (PCR) in 55 patients on chronic hemodialysis. Compared with HCV RNA by PCR, RIBA 3.0 increased the sensitivity of HCV detection to 72% as compared with 56% of RIBA 2.0. Both assays underestimated the prevalence of HCV infection as determined by PCR. However, RIBA HCV 3.0 outperformed RIBA HCV 2.0, detecting all of the RIBA 2.0-positive patients plus an additional eight (8 of 22 RIBA 2.0 negative; confidence interval [CI] = [17.2%, 59.3%]). Forty-three of 51 patients with positive RIBA 3.0 or positive HCV RNA by PCR underwent a liver biopsy. Thirty (70%) had chronic hepatitis (three with cirrhosis), 10 (23%) had nonspecific changes, and three (7%) had normal liver histology. Thirty of 37 patients (81%) with hepatitis C viremia and positive anti-HCV had chronic hepatitis, whereas none of the viremic patients with negative anti-HCV had chronic hepatitis. Among the reactive antigens on RIBA 3.0, c33c was found to be most predictive of chronic hepatitis (P = 0.0002). Detection of HCV RNA continues to be the method of choice in the early phase of HCV infection. In places where a validated HCV RNA assay is not available, RIBA HCV 3.0 (soon to be commercially available) is a better alternative. Early detection of HCV infection and the implementation of an isolation strategy might be important in preventing the spread of HCV infection among hemodialysis patients.(ABSTRACT TRUNCATED AT 250 WORDS)

Isolation and HPLC of N-epsilon-lithocholyl lysine as its fluorescamine and dimethylaminoazobenzene isothiocyanate derivatives.

N-epsilon-lithocholyl lysine (NELL) is a component of tissue-bound lithocholic acid (TBL). The isolation of NELL from native protein sources was simulated by hydrolysis of lithocholyl-bovine serum albumin (BSA) (synthesized by coupling lithocholyl-N-hydroxysuccinimide to fatty acid-free BSA) by digestion with a mixture of 6N HCl-propionic acid at 70 C for 3 h under partial vacuum. NELL was isolated on a reversed phase Sep-Pak C18 column and converted to either a fluorophor with fluorescamine or to a chromophor with dimethylaminoazobenzene isothiocyanate for subsequent HPLC using appropriate fluorescence or UV/visible absorption detectors. The procedure described here is quantitative, highly sensitive, and not dependent upon the use of Clostridial cholanoylamino acid hydrolase, the activity of which is sometimes blocked by steric hindrance on the substrate. Using this procedure we have demonstrated the presence of TBL in native histones.

Processing of envelope polypeptides of herpes simplex virus type 1. Demonstration of variation in different cell lines by high-performance liquid chromatography and radioimmunoprecipitation.

[35S]Methionine-labelled envelope polypeptides of herpes simplex virus type 1, strain F, propagated in mammalian cell culture of various origins, were separated by ion-exchange high-performance liquid chromatography on a TSK DEAE-3SW column. Analysis of the fractions by radioimmunoprecipitation followed by sodium dodecyl sulphate polyacrylamide gel electrophoresis of the immunoprecipitates showed similarities as well as distinct differences in the number, migration patterns and molecular mass of the synthesized polypeptides, depending on the host cell. The results show that this method can be used to demonstrate species-specific or organ-specific differences in the processing of virus-specified polypeptides synthesized in host cells.

Chorea as a presentation of herpes simplex encephalitis relapse.

Three infants, ages 3 months to 3 years, presented with chorea as the initial manifestation of herpes simplex encephalitis (HSE) relapse. Patient 2, treated with repeated 10 day courses of 30 mg/kg/day of acyclovir, had no clear improvement in neurological status. Patient 1, treated with a repeated 10-day course, improved only to have another HSE relapse 4 years later. Patient 3 clearly improved soon after a 3-week course of acyclovir at conventional dosages. A fourth patient (Patient 4) who relapsed with chorea after what was thought to be HSE, and who did not respond to repeated acyclovir treatment, was negative for herpes simplex virus indicators on brain biopsy and DNA testing. We recommend treating all patients suffering from HSE with a minimum 3-week course of acyclovir at 30-35 mg/kg/day in 3 divided doses.

Detection of herpes simplex virus DNA in cerebrospinal fluid of Saudi pediatric patients with encephalitis by the polymerase chain reaction.

Twelve samples of cerebrospinal fluid (CSF) from Saudi pediatric patients with suspected encephalitis were found negative for herpes simplex virus by cell culture and by nucleic acid hybridization using(32)P-labeled specific probes. After amplification of DNA extracted from 100 microl of each sample using primers for the glycoprotein D gene of HSV, seven specimens were found positive for HSV DNA. Subsequent hybridization with the labeled DNA probe ensured the positivity of the seven samples and further detected one more positive sample, for a total of eight HSV DNA positive cases. This technique improved the detection of HSV in CSF samples and may assist in an early diagnosis for HSV ecephalitis.

Genomic variation among herpes simplex virus type 1 strains: virus DNA analysis of isolates from Saudi patients.

Fifty-two clinical isolates of herpes simplex virus type 1 (HSV-1) from Saudi Arabian patients were analysed by restriction endonuclease digestion of the virus DNA using the enzymes HindIII and BamHI, followed by hybridization with 32P labelled DNA of laboratory strain F. Of the isolates, 17 were resolved into four distinct cleavage patterns with HindIII restriction enzyme. The remaining 35 strains had the same cleavage pattern as the standard HSV-1-F. Further investigation of the 52 isolates with BamHI, which is a multicut enzyme and therefore capable of higher resolution, differentiated 47 of the 52 isolates and were assigned into nine cleavage groups. Comparing our findings with similar studies reported elsewhere suggest geographic clustering of HSV-1 strains. Fragments giving rise to the observed DNA polymorphism were mapped to the unique region of the long and short segments of the genome.

Effect of lithocholic acid feeding on plasma lipoproteins and binding of radioiodinated human lipoproteins to hepatic membranes in rats.

1. Male Sprague-Dawley rats fed diets containing 0.25% lithocholic acid for 6 weeks exhibited elevated serum cholesterol. 2. The rats were fed diets containing 5 or 20% fat with and without the lithocholate and/or oxytetracycline-HCl. 3. The cholesterol elevation was associated with high density lipoprotein (HDL) and not very low density lipoprotein (VLDL) or low density lipoprotein (LDL). 4. Specific binding of human [125I]HDL to hepatic membranes was lowered in lithocholate-fed rats, but binding of human [125I]LDL to these membranes was not affected.