Healthcare-associated COVID-19 outbreaks: a nationwide population-based cohort study.

BACKGROUND: Healthcare-associated coronavirus disease 2019 (COVID-19) has significant implications for patients, their companions and healthcare workers (HCWs). Controlling transmission in healthcare settings is critical to reduce deaths due to COVID-19. AIM: To describe the epidemiology and characteristics of healthcare-associated COVID-19 outbreaks and outbreak-related cases. METHODS: The investigation data for each healthcare-associated outbreak that occurred between 15th January 2020 and 31st July 2021 in Taiwan were analysed retrospectively. Confirmed outbreak-associated cases were categorized as HCW cases, patient companion cases or patient cases, and the characteristics of the confirmed cases were compared between these categories. FINDINGS: In total, 54 healthcare-associated COVID-19 outbreaks including 512 confirmed cases were reported. The median number of affected cases per outbreak was six [interquartile range (IQR) 2-12], and the median outbreak duration was 12 days (IQR 4.3-17.0). Only 5.7% and 0.2% of all confirmed cases were partially and fully vaccinated, respectively. Most outbreaks (90%, 48/54) occurred in May and June 2021. HCW cases, companion cases and patient cases accounted for 19.5%, 41.2% and 39.3% of the total cases. Patient cases were significantly older (median age 72 years, IQR 61-83) and had higher 30-day all-cause mortality (37.4%) than HCW cases (median age 41 years, IQR 28-58, 0%) and companion cases (median age 52 years; IQR 42-62, 1%). CONCLUSION: Healthcare-associated COVID-19 outbreaks have a critical impact on patients. Nevertheless, two-thirds of cases in the healthcare-associated outbreaks in this study comprised HCWs and companions. In order to effectively mitigate COVID-19 transmission in healthcare settings, multi-pronged infection prevention and control measures should be implemented and tailored for these three groups.

Epidemiology of Bartonella infection in rodents and shrews in Taiwan.

During the period of August 2002 and November 2004, an epidemiological investigation for Bartonella infection was conducted in small mammals in Taiwan. Using whole blood culture on chocolate agar plates, Bartonella species were successfully isolated from 41.3% of the 310 animals tested. The isolation rate of Bartonella species varied among different animal species, including 52.7% of the 169 Rattus norvegicus, 28.6% of the 126 Sucus murinus, 10% of the 10 Rattus rattus and 66.7% of the three Rattus losea. Bacteremia prevalence also varied with the origin of the animals, as 56.2% of the animals captured on farms, 38.6% of the ones captured at harbour sites and 11.8% of the animals captured from urban areas were bacteremic. Through molecular analysis of the gltA gene and 16S/23S intergenic spacer region, genetic diversity of Bartonella organisms was identified, including strains closely related to Bartonella tribocorum, Bartonella grahamii, Bartonella elizabethae, Bartonella phoceensis and Bartonella rattimassiliensis. Moreover, this is the first report of zoonotic B. elizabethae and B. grahamii identified in R. losea, the lesser rice-field rat. Various Bartonella species were identified in R. norvegicus, compared to 97.2% of Suncus murinus with unique Bartonella species. By indirect immunofluorescence antibody test, using various rodent Bartonella species as antigens, consistently low percentage of seropositivity implied that small mammals may play a role as competent reservoirs of Bartonella species in Taiwan. Future studies need to be conducted to determine whether these Bartonella species would be responsible for human cases of unknown fever or febrile illness in Taiwan, especially zoonotic B. elizabethae and B. grahamii.

Measles seroepidemiology and decay rate of vaccine-induced measles IgG titers in Taiwan, 1995-1997.

Community-based seroepidemiologic studies were conducted to monitor the effectiveness of measles immunization programmes and to estimate the decay rate of vaccine-induced measles IgG titres. Sera collected from a mountain (792 sera), rural (875 sera) and urban (894 sera) populations in 1995-1997 were available. Measles IgG was quantified using a commercial EIA kit. Measles IgG seroprevalence and geometric mean titre (GMT) were calculated by setting the cut-off titre at 50 mIU/ml. The decay rate of measles IgG titres was estimated by assuming that the measles IgG titres, without exposing to wild measles virus, decay exponentially and constantly after 1 year post vaccination. The half-life of measles IgG titres was calculated from the corresponding decay rate. Measles IgG seroprevalences in these three populations have reached >95% in school children (7-18 years old) and >98% in young adults (19-25 years old) but varied from 87 to 96% in pre-school children (4-6 years old). Two-dose vaccinees, comparing with 1-dose vaccinees, had a significantly higher seroprevalence (98 versus 92%, P<0.01) and a slightly longer half-life of measles IgG titres (61 versus 27 months, P=0.08) but the measles IgG GMT in the two groups did not differ significantly (675 versus 618 mIU/ml, P=0.78).

Antibody to the nonstructural protein NS1 of Japanese encephalitis virus: potential application of mAb-based indirect ELISA to differentiate infection from vaccination.

An indirect enzyme-linked immunosorbent assay (ELISA) was developed to detect and differentiate the antibody responses to Japanese encephalitis (JE) virus nonstructural protein NS1 between infected and vaccinated individuals. The results showed that all convalescent sera from JE patients contained NS1-specific IgG antibodies, while 65 and 40% of these sera showed detectable NS1-specific IgM and IgA antibodies, respectively. Specificity analysis showed that NS1-specific IgM and IgA antibodies from JE patients do not cross-react to dengue virus NS1 glycoprotein, while IgG antibodies from 10% of JE patients showed significant cross-reaction to dengue virus NS1 glycoprotein. To differentiate infection from vaccination, the immune sera from 24 children vaccinated with inactivated JE vaccine were analyzed. The data showed that none of these immune sera had detectable NS1-specific IgG antibodies. The results demonstrated the potential application of JE NS1-specific indirect ELISA to differentiate infection from vaccination.

Dengue NS1-specific antibody responses: isotype distribution and serotyping in patients with Dengue fever and Dengue hemorrhagic fever.

To understand the antibody responses to dengue (DEN) nonstructural 1 (NS1) glycoprotein and their roles in protective immunity or pathogenesis of dengue fever (DF) and dengue hemorrhagic fever (DHF), we have analyzed the NS1-speccific IgM, IgA and IgG antibodies from patients with DF and DHF. An isotype-specific, indirect enzyme-linked immunosorbent assay (ELISA) was established by coating a NS1-specific monoclonal antibody (MAb), D2/8-1, to capture soluble NS1 antigens secreted in the culture supernatants of Vero cells infected with DEN virus. We observed strong anti-NS1 antibody responses in all of the convalescent sera of patients with DF and DHF. Similar NS1-specific isotypic and serotypic antibody responses were found in the sera from DF and DHF patients. The results showed that all DEN infections induced significant NS1-specific IgG, whereas 75% and 60% of primary DF patients vs. 40% and 90% of secondary DF patients produced IgM and IgA antibodies, respectively. Specificity analysis showed that DEN NS1-specific IgG and IgA antibodies cross-react strongly to Japanese encephalitis (JE) virus NS1 glycoprotein, whereas DEN NS1-specific IgM antibodies do not cross-react to JE virus NS1 glycoprotein at all. The serotype specificity of NS1-specific IgM, IgA and IgG were found to be 80%, 67% and 75% for primary infections, and 50%, 22% and 30% for secondary infections in positive samples of DF patients. Similar pattern was found in DHF patients. The results showed that all of the DF and DHF patients produced significant NS1-specific antibodies. We did not observe direct correlation between the anti-NS1 antibody responses and DHF because sera from patients with DF and DHF showed similar anti-NS1 antibody responses.

Significance of carbohydrate antigen sialyl-Lewis X, sialyl-Lewis A, and possible unknown ligands to adhesion of human urothelial cancer cells to activated endothelium.

OBJECTIVE: To assess the contribution of the carbohydrate antigens, sialyl-Lewis X (sLe(x)) and sialyl-Lewis A (sLe(a)), which are known to be ligands for E-selectin, to the adhesion between human urothelial cancer cells and cytokine-activated human endothelial cells. MATERIALS AND METHODS: We studied the expression of sLe(x) and sLe(a) antigens of three bladder cancer cell lines (JTC 30, JTC 32, and T24) by flow cytometry and the adherence to interleukin 1beta-activated human umbilical vein endothelial cells (HUVEC). RESULTS: JTC 30 and JTC 32 cells expressed both sLe(x) and sLe(a) antigens, and showed adhesion to activated HUVEC, which was completely abolished by anti-E-selectin antibody. T24 cells expressed neither sLe(x) nor sLe(a) antigen, and did not adhere to activated HUVEC. Each of anti-sLe(a) or anti-sLe(x) antibody partially blocked the attachment of JTC 30 cells to activated HUVEC, and combination of these antibodies almost completely blocked the adhesion. The combination of antibodies did not significantly influence the adhesion of JTC 32 cells. CONCLUSION: These results indicate that both sLe(a) and sLe(x) carbohydrate antigens are involved in E-selectin-mediated adhesion of some urothelial cancers, and that there might be unknown ligands for E-selectin on urothelial cancer cells.

Differentiation-induced transmigration of HL60 cells across activated HUVEC monolayer involves E-selectin-dependent mechanism.

The leukocyte-endothelial adhesive interaction is one of the key mechanisms during inflammation. The human promyelocytic cell line HL60 has been used in a number of studies to characterize leukocyte-endothelial interactions, especially selectin-mediated adhesion. HL60 also has been used in studies to characterize the myeloid cell function during differentiation. In this study, we investigated the adhesive interactions of HL60 to vascular endothelium, either in its undifferentiated state or after dimethylsulfoxide-induced granulocytic differentiation. Granulocytic differentiation of HL60 cells significantly enhanced their transmigration across cytokine-activated (IL-1 beta 10 U/ml, 4 h) HUVEC monolayer. Interestingly, this enhanced transmigration of differentiated HL60 cells was inhibited by pretreatment of the monolayers with anti-E-selectin mAb as well as anti-ICAM-1 mAb or anti-VE-cadherin mAb, suggesting a potential role for E-selectin in transendothelial migration. Further study of this enhanced transmigration mechanism may elucidate the regulation of selectin-mediated leukocyte-endothelial interactions.

The epidemiology of Japanese encephalitis on Taiwan during 1966-1997.

Japanese encephalitis (JE) is an endemic disease in Taiwan. A mass vaccination program of children against JE was first implemented in 1968. Along with general improvements in various aspects of living conditions over the years, the program has brought JE well under control. The main characteristics of JE epidemiology in Taiwan in the past 3 decades are as follows. The transmission mode remains unchanged-that is, the amplification stage of the virus in pigs is followed by a human epidemic each year. The frequency of JE incidence has dropped significantly. The incidence rate of confirmed cases was 2.05 per 100,000 in 1967, the highest in record, and merely 0.03 per 100,000 in 1997. Confirmed cases occur sporadically all over the island. The peak of the epidemic season has shifted from August in the 1960s to June since the 1980s. The age distribution of confirmed cases has shifted gradually from mainly children to adults. Vaccine efficacy for those having received more than 2 doses of the vaccine is estimated to be about 85%.

Antibody responses to an immunodominant nonstructural 1 synthetic peptide in patients with dengue fever and dengue hemorrhagic fever.

Two flaviviruses, dengue (DEN) virus and Japanese encephalitis (JE) virus, are important because of their global distribution and the frequency of epidemics in tropical and subtropical areas. To study the B-cell epitopes of nonstructural 1 (NS1) glycoprotein and anti-NS1 antibody response in DEN infection, a series of 15-mer synthetic peptides from the predicted B-cell linear epitopes of DEN-2 NS1 protein were prepared. Enzyme-linked immunosorbent assay (ELISA) was performed to analyze antibody responses to these peptides from sera of both DEN and JE patients. One peptide derived from DEN-2 NS1, D2 NS1-P1 (amino acids 1-15), was identified as the immunodominant epitope that reacted with sera from dengue fever (DF) patients but not JE patients. The isotype of D2 NS1-P1-specific antibodies was mainly immunoglobulin M (IgM) in all sera that tested positive. A specificity study demonstrated that sera from all four DEN types reacted with D2 NS1-P1. A dynamics study showed that specific antibodies to this peptide could be detected as early as 2 days after the onset of symptoms. We observed significant anti-D2 NS1-P1 antibody responses in 45% of patients with primary and secondary infections with DF or with dengue hemorrhagic fever. This is the first report demonstrating that significant anti-DEN NS1 antibodies can be induced in the sera of patients with primary DEN infection.

RNA-protein interactions: involvement of NS3, NS5, and 3' noncoding regions of Japanese encephalitis virus genomic RNA.

The mechanism of replication of the flavivirus Japanese encephalitis virus (JEV) is not well known. The structures at the 3' end of the viral genome are highly conserved among divergent flaviviruses, suggesting that they may function as cis-acting signals for RNA replication and, as such, might specifically bind to cellular or viral proteins. UV cross-linking experiments were performed to identify the proteins that bind with the JEV plus-strand 3' noncoding region (NCR). Two proteins, p71 and p110, from JEV-infected but not from uninfected cell extracts were shown to bind specifically to the plus-strand 3' NCR. The quantities of these binding proteins increased during the course of JEV infection and correlated with the levels of JEV RNA synthesis in cell extracts. UV cross-linking coupled with Western blot and immunoprecipitation analysis showed that the p110 and p71 proteins were JEV NS5 and NS3, respectively, which are proposed as components of the RNA replicase. The putative stem-loop structure present within the plus-strand 3' NCR was required for the binding of these proteins. Furthermore, both proteins could interact with each other and form a protein-protein complex in vivo. These findings suggest that the 3' NCR of JEV genomic RNA may form a replication complex together with NS3 and NS5; this complex may be involved in JEV minus-strand RNA synthesis.

Silent transmission of the dengue virus in southern Taiwan.

Immunoglobulin M (IgM) antibody to dengue virus was examined from a total of 3,099 serum samples collected in southern Taiwan. Of 1,232 sera collected from a junior high school and four elementary schools in Liu-Chiu, 35 were IgM-positive, demonstrating that the dengue virus has been circulating on the island, despite the fact that no epidemic has been reported in the past 10 years. Sixteen of 925 sera collected from three elementary schools in Tung-Kang in 1991 were found to be IgM-positive and two of 192 sera from adults in the local community were positive. The IgM-positive subjects tended to be aggregated around a port. Fishing boats that had stopped in neighboring endemic countries were presumed to have introduced the virus periodically, causing a low level of inapparent infections. In the Kaohsiung area, two of 108 suspected clinical cases and four of 642 community-based sera were IgM-positive. Rapid urbanization has provided appropriate circumstances for vector breeding in this area and the high population density has also increased contact frequency between humans and mosquito vectors. This has, in turn, increased the possibility of silent transmission of the dengue virus via either intermittent reintroduction of the virus or continuation of inapparent infections or both. Establishment of a early warning system using the IgM antibody capture-enzyme-linked immunosorbent assay is suggested for effective monitoring of the disease.

Identification of immunodominant, group-specific and subcomplex-specific, continuous epitopes in the core regions of Japanese encephalitis virus using synthetic peptides.

Two flaviviruses, Japanese encephalitis (JE) virus and Dengue (DEN) virus which have high pathogenicity for humans, continue to pose a serious public health problem in tropical and subtropical countries of the world. In order to identify the immunodominant B-cell epitopes for diagnostic application, we have prepared a series of 15-mer synthetic peptides from JE virus core protein based on computer analysis. Four linear, immunodominant epitopes corresponding to amino acids 91-105 (P78), 1-15 (P73), 8-22 (P74), and 34-48 (P75) of JE virus core proteins were identified by employing an enzyme-linked immunosorbent assay (ELISA), using high-titered immune sera from JE-vaccinated children. P78 was found to be the most immunodominant. The sero-specificity of these peptides was tested by binding to seroconverted samples from JE and DEN-1 patients. P78 and P74 belonged to group-specific epitopes which reacted with both JE and DEN-1 patient sera. P73 and P75 belonged to subcomplex-specific epitopes which reacted only with JE but not with DEN-1 patient sera. The study suggests that these peptides corresponding to the immunodominant epitopes of JE virus core protein might have the potential to be used as peptide-based diagnostic reagents for the detection and differentiation of JE and DEN antibody responses.

Clinical efficacy of recombinant human erythropoietin in the treatment of anemia in hemodialysis patients: influence of dosing regimen, iron status, and serum aluminum.

To evaluate the clinical efficacy of recombinant human erythropoietin (EPO) and its influencing factors in the treatment of anemia in hemodialysis (HD) patients, 17 chronic stable HD patients (10 males, 7 females; mean age: 46.0 +/- 2.6 years) with severe anemia were enrolled in this study. The study period (ranging from 5 to 11 months) was divided into the initial 12 weeks of correction phase and the subsequent maintenance phase. EPO, 1500 U initially, was administered intravenously twice weekly (BIW group, n = 10) or thrice weekly (TIW group, n = 7) at the end of each HD. Dose was doubled every 4 weeks until up to a maximum dose of 6000 U if increment of hematocrit (Hct) was less than 3%. At the end of correction phase, anemia was markedly improved. Hct and hemoglobin (Hb) increased from 19.3 +/- 0.8 to 28.7 +/- 1.1% and from 6.5 +/- 0.3 to 9.6 +/- 0.4 g/dl, respectively. Fifteen patients (88%) reached to the target Hct of 30% at 13.7 +/- 1.2 weeks. At the end of study, Hct and Hb was maintained at 29.1 +/- 0.7% and 9.6 +/- 0.3 g/dl, respectively. Requirement of EPO dose to reach the target and maintain the stable Hct (greater than or equal to 28%) was 99 +/- 14 and 62 +/- 11 U/kg/week, respectively. Laboratory parameters showed that serum iron, transferrin saturation, sugar and triglyceride decreased significantly and uric acid and aluminum (Al) increased significantly. There was no significant change in predialysis blood pressure, body weight, cardiac ratio, and ECG. Quality of life was markedly improved with the better subjective feelings, physical activity and Karnorfsky index. Common adverse effects included exacerbated hypertension (23%), hyperphosphatemia (18%), hyperkalemia (18%), and flu-like syndrome (12%). All of them could be managed by medical and dialysis treatment. Investigation of influencing factors on response to EPO suggests that 1) TIW group had a better response than BIW group 2) Response was better in patients with more adequate iron status and less severe Al burden. 3) Time to target Hct correlated approximately with basal serum Al levels but did not correlate with basal serum parathyroid hormone levels. In conclusion, low dose of EPO therapy corrects anemia effectively with minimal adverse effects in HD patients. Dosing regimen, iron status, and serum Al will influence the response to EPO.