Suboptimal MMR Vaccination Coverages-A Constant Challenge for Measles Elimination in Romania.

Measles is targeted for elimination since 2001, with a significant reduction in cases recorded worldwide, but outbreaks occur periodically due to immunization gaps. This study analyzes the evolution of vaccination coverage rates (VCRs) in Romania, a EU country with large measles epidemics during the last two decades, including an ongoing outbreak in 2023. Vaccination against measles has been part of the National Immunization Program since 1979, initially as a single dose, and from 1994 onwards it has had two doses. The initially high national VCRs of >97% gradually declined from 2010 onward and remained constantly under 90%, with further decreases during the COVID-19 pandemic. The lowest VCRs for both vaccine doses in the last decade were recorded in 2022 and were 83.4% for the first dose and 71.4% for the second dose, with significant differences among Romania's 42 counties. Several factors contributed to this decline, including failure to attend the general practitioners' offices, increased number of children lost to follow-up due to population movements, missed vaccination opportunities due to temporary medical contraindications, a surge in vaccine hesitancy/refusal, a decreasing number of general practitioners and discontinuities in vaccine supply. The persisting suboptimal VCRs in Romania threaten the progress toward measles elimination.

HIV-HBV Coinfection-Current Challenges for Virologic Monitoring.

HIV-HBV coinfected patients have higher rates of liver-related morbidity, hospitalizations, and mortality compared to HBV or HIV mono-infected ones. Clinical studies have shown an accelerated progression of liver fibrosis and an increased incidence of HCC, resulting from the combined action of HBV replication, immune-mediated hepatocytolysis, and HIV-induced immunosuppression and immunosenescence. Antiviral therapy based on dually active antiretrovirals is highly efficient, but late initiation, global disparities in accessibility, suboptimal regimens, and adherence issues may limit its impact on the development of end-stage liver disease. In this paper, we review the mechanisms of liver injuries in HIV-HBV coinfected patients and the novel biomarkers that can be used for treatment monitoring in HIV-HBV coinfected persons: markers that assess viral suppression, markers for liver fibrosis evaluation, and predictors of oncogenesis.

HIV transmission clusters among injecting drug users in Romania.

Injection drug use is increasingly an important route of HIV transmission in Romania (from 1.5% of the newly diagnosed cases prior to 2010 to 31% in 2013). In this study we investigated the viral characteristics and relationships in newly HIV infected persons who inject drugs in Bucharest, Romania. RESULTS: HIV-1 pol sequencing, followed by phylogenetic and clustering analysis was performed on blood from 37 injecting drug users (IDUs) newly diagnosed with HIV infection. While HIV subtype F1, the dominant strain in Romania since 1990, remains prevalent, new subtypes were found including G, B, B/G and B/F recombinants. Overall, 27 of the available sequences (72.9%) clustered with at least one other. Network and phylogenetic analysis revealed tight monophyletic clusters for both subtypes F and G, with short genetic distances between sequences, suggesting recent numerous acute to acute transmissions or single burst-type episodes. No transmitted drug-resistance mutations were identified. Greater immunosuppression was present in subjects forming the subtype G cluster, possibly indicating a faster rate of progression associated with this subtype. CONCLUSIONS: The recent increasing numbers of IDU related HIV transmissions in Bucharest, has resulted in closely-knit transmission networks that maychange the genetic profile of the local HIV epidemic.

HCV non-1b genotypes in injecting drug users from Romania.

INTRODUCTION: Chronic hepatitis C cases diagnosed in Romania were mostly related to unsafe parenteral treatments and blood transfusions; HCV genotype 1b was prevalent. During the last decade, an increasing number of HCV infections was reported among people who inject drugs (PWID). The aim of the current study was to test if this epidemiological shift triggered a diversification of the circulating viral strains. METHODOLOGY: HCV genotypes were determined by reverse hybridization in 130 HCV-infected PWID (87.7% males; mean age 27.9 ± 6.7 years, injecting drugs for 8.1 ± 4.8 years). RESULTS: HIV-HCV co-infection was diagnosed in 80.8% of the subjects and 26.9% were HIV-HCV-HBV triple infected. Active HCV viral replication was present in 104 PWID (80%), more frequently in those HIV-co-infected (91.4% vs. 52% in HCV mono-infected, and 77.148.5% in HIV-HCV-HBV triple-infected, p = 0.0001). Non-1b genotypes were prevalent (54.8%), with subtype 1a the most commonly detected (24%), followed by genotypes 3a (14.4%) and 4 (7.7%). Mixed infections with genotypes 1a and 1b were found in nine subjects (8.7%). There was no difference in the genotypes frequencies based on HIV or HBV co-infection status, length of drug usage, or associated risk factors (tattoos, piercing, detention). CONCLUSION: The continuous surveillance of HCV genotypes in PWID from Romania will add valuable information to the overall European epidemiological picture, with important therapeutic implications.

Injecting drug use: A vector for the introduction of new hepatitis C virus genotypes.

Hepatitis C virus (HCV) genotypes' monitoring allows real-time insight into the dynamic changes that occur in the global epidemiological picture of HCV infection. Intravenous drug use is currently the primary driver for HCV transmission in developed and developing countries. The distribution of HCV genotypes/subtypes differs significantly between people who inject drugs (PWID) and the general population. HCV genotypes that previously exhibited a limited geographical distribution (3a, 4) are becoming more prevalent in this high-risk group. Immigration from HCV-endemic countries and the evolving networks of HCV transmission in PWID influence HCV genotypes distribution in Europe. Social vulnerabilities (e.g., unemployment, homelessness, and limited access to social and healthcare insurances systems) are important triggers for illicit drug use, which increases the associated risks of HCV infection and the frequent emergence of less prevalent genotypes. Genotype/subtype determination bears important clinical consequences in the progression of liver disease, susceptibility to antiviral therapies and the emergence of resistance-associated variants. An estimated half of the chronically HCV-infected PWID are unaware of their infection, and only one in ten of those diagnosed enter treatment. Nevertheless, PWID exhibit high response rates to new antiviral regimens, and the level of HCV reinfection is unexpectedly low. The focus of the healthcare system must be on the early detection and treatment of infection, to avoid late presentations that are associated with high levels of viremia and liver fibrosis, which may diminish the therapeutic success rate.

Influence of Staphylococcus aureus attachment to the herpes simplex virus infected cells.

The purpose of this study was to investigate the response of HeLa cells to the interaction with inactivated Staphylococcus aureus cells and live challenge with herpes simplex virus (HSV).The results of this study are indicating that the interaction between the HeLa cells and S. aureus inactivated whole cells could modulate the host cell apoptosis and cytokine production, and therefore, influence the progression of HSV infection. The pre-treatment of HeLa cells with heat inactivated bacterial whole cells protects them from the occurrence of HSV mediated cytopathic effect, while the post viral infection treatment with bacterial cells prevents the high activation of bax/bcl-2 apoptotic pathway, a process that could change the fate of the infectious process triggered by the virus, and eventually reduce its multiplication rate. The pre-treatment of HeLa monolayer with inactivated bacterial cells 24 hours before the viral infection is increasing the expression level of TNF-a, IL-6 and IL-8 pro-inflammatory cytokines genes, also suggesting that bacterial antigens could contribute to the decrease of viral multiplication rate.

ROMANIAN SCIENTISTS IN THE NOMINATION DATABASE FOR THE NOBEL PRIZE IN PHYSIOLOGY OR MEDICINE, 1901-1951.

Nobelprize.org site is the most reliable and complete resource of information on the Nobel Prize and the Nobel Laureates. The nomination database for Physiology or Medicine, 1901-1951, offers exciting facts about the Romanian Schools of Medicine from Bucharest, Iasi and Cluj. Between 1920-1950, four Romanian scientists were nominated for the Nobel Prize: Victor Babes (1854-1926), Ion Cantacuzino (1863-1934), Thoma Ionescu (1860-1926) and Constantin Levaditi (1874-1953). This paper discusses these nominees, the nominators and the motivations, as well as the specific publications that endorse the candidates' scientific activity. Recommendations made by Romanian professors for foreign researchers to receive the Nobel Prize are also included.

The effectiveness of cytological rescreening in the reduction of false negative/positive Pap reports.

BACKGROUND: Cytological investigation of the cervix has proven to be a valuable tool in the early detection of cervical cancer; however, the high incidence of false negative or false positive smear reports is an important drawback. OBJECTIVES: To investigate retrospectively the value of partial rescreening methods as tools for improving the sensitivity and specificity of Pap test routine screening. METHODS: Out of a total of 4664 cervical samples examined by Pap test, 20% were randomly selected and rescreened with a more detailed examining protocol by the same cytologist; in addition, targeted rescreening of all samples with severe lesions was carried out. RESULTS: During initial testing, 478 smears (10.24%) showed cytological abnormalities, classified as ASC-US (5.79%); L-SIL (3.32%) and H-SIL (1.14%). At random rescreening, a significant decrease in the number of negative smears (83.05% vs. 85.9%, p = 0.036) was recorded, together with an increase (7.68% vs. 5.79%, p = 0.043) in the number of smears classified as ASC-US. No significant differences were recorded for L-SIL or H-SIL samples. Retrospective targeted rescreening of all 208 samples initially diagnosed as L-SIL and H-SIL revealed 42 false positive results and 12 false negative ones. Errors were linked to suboptimal smear preparation: scant cellularity, material in clumps, paucity of abnormal cells, pale dyskarosis, small microbiopsy- like aggregates. CONCLUSION: Partial random rescreening or targeted rescreening enables a better interpretation of suboptimal prepared smears. Targeted rescreening allows a correct detection of even low percentages of atypical cells. Other confounding factors, such as the laboratory workload and the regional prevalence of the disease, can exert an important effect on the correct classification of cytological lesions.

Investigation of Th1/Th2 cytokine profiles in patients with laryngo-pharyngeal, HPV-positive cancers.

The aim of this study was to reveal the relationships between the features of the primary tumour, the degree of tumour stage, the presence of human papillomaviruses (HPV) in blood and the severity of Th1/Th2 serum cytokine imbalance in patients with laryngo-pharyngeal cancer. The study was performed on 50 patients (47 men and 3 women), with age ranging from 40 to 83 years (the mean of the patients' ages was 58.4 ± 9.43 years, with a median of 60 years). A control group was represented by age-matched healthy patients (with no clinical diseases). The viral DNA was detected by PCR; the cytokine levels were determined by ELISA. A clear switch from cytokine Th1 to cytokine Th2 in cancer patients, low levels of IL-2 and IFNγ in advanced stages, as well as a positive correlation of increased levels of both IL-2 and IL-12 with the early stages of laryngo-pharyngeal cancer was observed. Loco-regional metastases were correlated with increased levels of IL-8 and IL-10 and drastic decrease of IFNγ. In advanced cancer stages, we found that the most affected were IL-2 and IFNγ correlated with increased levels of Th2 cytokines. Patients with HPV present in both primary tumours and blood showed increased values of IL-4:IL-2 ratio as compared with patients with HPV-positive primary tumours only, demonstrating the aggravation of the immunosuppressive state. The most important finding of our study is that for a correct evaluation of the Th1 to Th2 switch in cancer patients, it is necessary to establish not only the negative/positive correlations between different Th1 and Th2 type cytokines, but also the ratio between them. These parameters allowed us to state that the presence of HPV DNA in blood was associated with the most severe immunological imbalance that could potentially lead to a poor prognosis of these patients. Our findings encourage us to consider that the ratio between different Th1 and Th2 cytokines could represent a useful marker for clinical and pathological evaluation of cancer patients.

New interferons in the treatment of chronic hepatitis C.

The current standard therapy for chronic HCV infection is a combination of pegylated-interferon (PEG-IFN) and weight-based ribavirin, administered for 24-48 weeks, according to the viral genotype. Although the weekly administration of pegylated interferons provides superior antiviral efficacy over standard interferon alpha, the rate of sustained virological response rarely overpasses 50% in patients infected with HCV genotypes 1 and 4. Consequently, multiple clinical trials with congeners of interferon (consensus interferon, interferon lambda, albinterferon, and controlled-release interferons) are ongoing. Their main advantages consist in maintenance of viral suppression across a longer dosing interval, avoidance of interdose trough and reduced dosing frequencies (twice or even once per month compared to once per week for the actual PEG-IFNs). Along with these superior pharmacokinetic properties, new interferons are expected to have improved side-effect profiles and better tolerability compared with the currently available formulations, providing an option for otherwise difficult to treat, challenging populations. New interferon formulation can be incorporated into future combination with direct acting antivirals, in order to maintain viral suppression over longer periods and minimize the development of viral resistance.

Tracking down of laryngo-pharyngeal metastasis.

BACKGROUND: In Romania, the most optimistic statistics give a 5 years survival rate in approximately 33% of laryngo-pharyngeal cancer patients. Considering that a cell carrying the viral DNA is originating from primary tumor, we have tested whether HPV DNA could be detected in the blood cell of patients with laryngeal cancer as a marker of disease progression and metastases. METHODS: The study was performed on 85 patients (59 +/- 8.7 age) with laryngo-pharyngeal cancer. HPV DNA was detected in tumor using nested PCR with consensus primers, and also in local lymph nodes and/or blood cells from patients HPV positive in primary tumor. RESULTS: HPV DNA was detected in 75.29% of analyzed tumours, and all HPV16 positive samples were confirmed by mRNA E6 expression. 56.3% of patients presented HPV DNA in peripheral circulation as confirmed by PCR with E6 HPV16 specific primers followed by Southern Blot. CONCLUSION: Our results sustain that the detection of HPV DNA in blood is a "surrogate marker" of metastasis when extension of metastasis cannot be estimated, this observation is very important for management of cancer patients with laryngopharyngeal localization.

Influence of social changes on the evolution of HIV infection in Romania.

The Romanian HIV epidemic started and evolved as a paediatric one, accounting for more than a quarter of the total European juvenile AIDS cases. In response to this major AIDS outbreak, emphasis was placed on the patient's treatment, by implementation of a free, universal access program of Highly Active AntiRetroviral Therapy. This approach has been highly successful, and has greatly increased the rate of survival in infected children. Nevertheless, these children are now teenagers or young adults, representing a large cohort of "long term survivors"- a unique population that represent a great challenge for the public health system and for their integration in the civil society. As the number of HIV infected adults is increasing, new high-risk behaviour groups, as well as vulnerable populations (young people, people living in poverty, Rroma community) need to be reached in prevention programs. This article explores the impact of the socio-economic changes on the evolution of the HIV epidemic in Romania and speculates about the factors that might drive future increases in the incidence of HIV infections.

High prevalence of hepatitis B virus markers in Romanian adolescents with human immunodeficiency virus infection.

BACKGROUND: We evaluated the frequency of hepatitis coinfection in Romanian adolescents who were diagnosed with human immunodeficiency virus (HIV) infection prior to 1995. METHODS: One hundred sixty-one adolescents (13-18 years of age) with symptomatic HIV infection, but without signs of hepatic dysfunction, and 356 age-matched, HIV-uninfected controls underwent laboratory testing for markers of parenterally acquired hepatitis virus infection. RESULTS: Seventy-eight percent of HIV-infected adolescents had markers of past or present hepatitis B virus (HBV) infection, as compared with 32% of controls ( P = .0001). The prevalence of HBV replicative markers was more than 5-fold higher in HIV-infected adolescents as compared with controls: 43.4% vs 7.9% ( P = .0001), respectively, for hepatitis B surface antigen (HBsAg); and 11.2% vs 2.2% ( P = .0001), respectively, for hepatitis B e antigen (HBeAg). The prevalence of HBsAg chronic carriers and the presence of HBV replicative markers was significantly higher in patients with immunologically defined AIDS (CD4+ cell counts < 200 cells/mcL): 59.6% vs 34.6% ( P = .02) for HBsAg and 22.8% vs 5.7 %, ( P = .002) for HBV DNA. After 1 year of follow-up, the proportion of those who cleared the HBeAg was considerably lower in severely immunosuppressed coinfected patients: 4.7% vs 37.1% ( P = .003). Four additional HIV-infected adolescents became HBsAg-positive over the term of follow-up (incidence rate, 24.9/1000 person-years), despite a record of immunization against hepatitis B. CONCLUSIONS: A substantial percentage of HIV-infected and HIV-uninfected Romanian adolescents have evidence of past or present HBV infection. In HIV-infected adolescents, the degree of immunosuppression is correlated with persistence of HBV replicative markers, even in the absence of clinical or biochemical signs of liver disease.

Alternatives to HIV-RNA and CD4 count to monitor HIV disease progression: a prospective cohort study in Romania.

Little evidence exists on how to efficiently and effectively monitor HIV-disease progression in developing countries. Better understanding regarding cost-effective tests may help to resolve questions regarding treatment. A prospective cohort study was conducted with a 1-year follow-up period. Immune complex-dissociated (ICD) p24 antigen (ICD p24Ag), alone or in combination with HIV p24 antibody (p24Ab), was compared to HIV-RNA and CD4+ count in a cohort of 160 HIV-infected adolescents in Romania. The main outcome measure was disease progression, defined as >50,000 copies/ml of HIV-RNA or death. Among the 160 adolescents, a higher mean ICD p24Ag was significantly associated with clinical disease classification (CDC), plasma HIV-RNA concentration, and p24Ab. Multivariate logistic regression showed detectable ICD p24Ag had an odds ratio of 3.7 (95% CI 1.4-9.7) for disease progression in comparison to undetectable ICD p24Ag. ICD p24Ag is of value in determining the prognosis of disease in HIV-1-infected adolescents in developing countries. Additional studies for validation of this assay for HIV clades primarily affecting developing countries, are now needed.

High prevalence of hepatitis B virus markers in romanian adolescents with human immunodeficiency virus infection.

BACKGROUND: We evaluated the frequency of hepatitis coinfection in Romanian adolescents who were diagnosed with human immunodeficiency virus (HIV) infection prior to 1995. METHODS: One hundred sixty-one adolescents (13-18 years of age) with symptomatic HIV infection, but without signs of hepatic dysfunction, and 356 age-matched, HIV-uninfected controls underwent laboratory testing for markers of parenterally acquired hepatitis virus infection. RESULTS: Seventy-eight percent of HIV-infected adolescents had markers of past or present hepatitis B virus (HBV) infection, as compared with 32% of controls (P = .0001). The prevalence of HBV replicative markers was more than 5-fold higher in HIV-infected adolescents as compared with controls: 43.4% vs 7.9% (P = .0001), respectively, for hepatitis B surface antigen (HBsAg); and 11.2% vs 2.2% (P = .0001), respectively, for hepatitis B e antigen (HBeAg). The prevalence of HBsAg chronic carriers and the presence of HBV replicative markers was significantly higher in patients with immunologically defined AIDS (CD4+ cell counts < 200 cells/mcL): 59.6% vs 34.6% (P = .02) for HBsAg and 22.8% vs 5.7%, (P = .002) for HBV DNA. After 1 year of follow-up, the proportion of those who cleared the HBeAg was considerably lower in severely immunosuppressed coinfected patients: 4.7% vs 37.1% (P = .003). Four additional HIV-infected adolescents became HBsAg-positive over the term of follow-up (incidence rate, 24.9/1000 person-years), despite a record of immunization against hepatitis B. CONCLUSION: A substantial percentage of HIV-infected and HIV-uninfected Romanian adolescents have evidence of past or present HBV infection. In HIV-infected adolescents, the degree of immunosuppression is correlated with persistence of HBV replicative markers, even in the absence of clinical or biochemical signs of liver disease.

Cloning and expression of the L1 major capsid protein of the human papillomavirus 18 in murine cells.

Human papillomavirus (HPV) type 18 is strongly associated with the development of cervical cancer. Studies of a model system with animal papillomaviruses have demonstrated the importance of neutralizing antibodies in preventing papillomavirus-associated disease. The immune response to HPV is poorly understood, and there are non- standardized serological assays to identify HPV infections. In our study, the assessment of antibody responses against HPVs (previously hampered by the lack of viral source) was enabled by the expression of the L1 major capsid viral protein type 18 (HPV18) into L929 murine cells using the pTARGET mammalian expression vector system (MEVS). The cloning was validated by PCR with specific primers for the L1 gene, as well as by enzyme restriction and in situ hybridization. The evidence for the viral cloned gene expression was acquired by RT-PCR. Presence and antigenic properties of the recombinant L1 protein were shown using it as antigen in an indirect enzyme linked immunosorbent assay (ELISA) system. Significantly higher reactivity was noted when the sera samples were from persons infected with HPV18 as compared with the non-infected individuals but a moderately different reactivity was observed when the sera from patients infected with other HPV genotypes were tested. The results showed that the murine transfected cells could be used as antigen in order to detect the presence of the specific antibodies in HPV infected persons.