Long-term trends in HIV care entry: over 15 years of clinical experience from Poland.

INTRODUCTION: Delay in HIV diagnosis and consequently late care entry with low CD4 counts remain a major challenge for the control of the HIV/AIDS epidemic. The aim of this study was to analyse the evolution of characteristics of the HIV epidemic in Poland. METHODS: Cross-sectional data were collected for 3972 HIV-infected patients followed up in 14 of 17 Polish HIV treatment centres in the years 2000-2015. Clinical data were analysed and factors associated with late presentation (baseline CD4 count < 350 cells/µL or history of AIDS-defining illness) and advanced HIV disease (baseline CD4 count < 200 cells/µL or history of AIDS) were identified. RESULTS: The majority (57.6%) of patients entered care late, while 35.6% presented with advanced HIV disease. The odds of being linked to care late or with advanced HIV disease increased consistently across age categories, increasing from 2.55 [95% confidence interval (CI) 1.46-4.47] for late presentation and 3.13 (95% CI 1.49-6.58) for advanced disease for the 21-30-year-old category to 5.2 (95% CI 1.94-14.04) and 8.15 (95% CI 2.88-23.01), respectively, for individuals > 60 years of age. Increased risks of late entry and advanced HIV disease were also observed for injecting drug users [adjusted odds ratio (aOR) 1.74 (95% CI 1.16-2.60) and 1.55 (95% CI 1.05-2.30), respectively], with lower aOR associated with the men who have sex with men transmission route [aOR 0.3 (95% CI 0.31-0.59) and 0.39 (95% CI 0.29-0.53), respectively]. The frequencies of cases in which patients were linked to care late and with advanced HIV disease decreased over time from 67.6% (2000) to 53.5% (2015) (P < 0.0001) and from 43.5% (2000) to 28.4% (2015) (P = 0.001), respectively. CONCLUSIONS: Despite improvements over time, most patients diagnosed with HIV infection entered care late, with a third presenting with advanced HIV disease. Late care entry remains common among people who inject drugs and heterosexual groups.

High frequency of neurosyphilis in HIV-positive patients diagnosed with early syphilis.

BACKGROUND: Syphilis is an infection frequently seen with HIV, and European guidelines on the management of syphilis suggest that HIV-infected patients may have an increased risk of early neurological involvement, sometimes asymptomatic. Recent study shows a relationship between neurosyphilis and cerebrospinal fluid (CSF) HIV viral load (VL), which in turn may be associated with subsequent neurocognitive decline. OBJECTIVES AND METHODS: The aim of the study was estimation of the frequency of neurosyphilis among HIV-positive patients with early syphilis. The study included all patients diagnosed with early syphilis who had lumbar puncture performed in the years 2008-2012. Analysis included CSF parameters (serology, mononuclear cells, protein, glucose, chloride and lactate levels), CD4 count, serum VL and highly active antiretroviral therapy (HAART). Diagnosis of neurosyphilis was confirmed by CSF serology [positive fluorescent treponemal antibody and/or Venereal Disease Research Laboratory (VDRL) test(s)] and increased number of mononuclear cells. Statistical analysis included χ(2) tests with an accepted significance level of P < 0.05. RESULTS: Lumbar puncture was performed in 72 patients, all men, with median age 33 (interquartile range 11) years. Neurosyphilis was confirmed in 65 (90.28%) of the patients. No statistically significant association between CSF parameters and CD4 count was found. However, statistically significant associations were found only between pleocytosis and serum VL > 1000 HIV-1 RNA copies/mL (P = 0.0451), as well as HAART treatment (P = 0.0328). The proportion of confirmed neurosyphilis cases, also in patients with low serum VDRL titres, was very high. CONCLUSIONS: Considering the high proportion of patients who objected to having LP performed in the absence of neurological symptoms and the risk associated with this procedure, it may be preferable to use treatments with good CNS penetration in all HIV-positive patients with early syphilis.

Hepatitis C virus (HCV) genotype 1 NS5A resistance-associated variants are associated with advanced liver fibrosis independently of HCV-transmission clusters.

OBJECTIVES: The aim of the study was to characterize the differences in the frequencies of NS3 and NS5A resistance-associated variants (RAVs) among Polish therapy-naive genotype 1 (G1) hepatitis C virus (HCV)-monoinfected and human immunodeficiency virus (HIV)/HCV-coinfected patients including clustering patterns and association of RAV frequency with liver fibrosis. METHODS: NS3/NS5A RAVs were identified by population sequencing in 387 directly acting antiviral treatment-naive G1-infected individuals (54 with genotype 1a (G1a) and 333 with genotype 1b (G1b)). Liver fibrosis was assessed based on histopathology or ultrasound elastography. Phylogenetic clusters were identified using maximum likelihood models. For statistics, chi-squared or two-sided Fisher's exact tests and multivariate logistic regression models were used, as appropriate. RESULTS: NS3 RAVs were found in 33.33% (18/54) for G1a and 2.62% (8/297) for G1b whereas NS5A variants were present in 5.55% (3/54) G1a and 9.31% (31/333) G1b sequences. Variations in NS5A 31 and 93 codon positions were found only in G1b (4.2% (14/333) for L31I/F/M and 5.39% (17/333) for Y93H). NS5A RAVs were more frequent among patients with advanced liver fibrosis (17.17% (17/99) for F3-F4 versus 6.94% (17/245) for F0-F2; p 0.004) or liver cirrhosis (20.34% (12/59) for F4 versus 7.72% (22/285) for F0-F3; p 0.003). Liver cirrhosis (F4) was associated with higher odds ratio of the NS5A RAVs among HCV-infected patients (odds ratio 2.34, 95% CI 1.004-5.291; p 0.049). NS5A RAVs were less frequent among sequences forming clusters and pairs (5.16% (8/155) versus 11.21% (26/232); p 0.039). CONCLUSIONS: Presence of NS5A RAVs correlated with progression of liver fibrosis and represents de novo selection of variants rather than transmission of drug resistance. Hence, the presence of NS5A RAVs may be a predictor for a long-lasting HCV infection.

Co-receptor usage was more predictive than NSI/SI phenotype for HIV replication in macrophages: is NSI/SI phenotyping sufficient?

A monocyte-derived macrophage (MDM) culture assay was used to define the replication kinetics of HIV isolates. Ten-day-old MDMs were infected with HIV. Supernatants were collected and assayed for HIV p24 on days 3, 7, 10, and 14 post-infection (PI). In this assay, SF162 (macrophage tropic, NSI) produced increasing amounts of HIV p24 antigen with increasing time in culture. BRU (nonmacrophage tropic, SI) infection resulted in low levels of HIV p24 antigen with no increase in production during the culture period. A panel of 12 clinical isolates was evaluated. All isolates produced detectable levels of HIV p24 antigen in MDMs. However, the NSI viruses had significantly higher log10 HIV p24 antigen values at all times PI (P < 0.01). Co-receptor usage was determined for all 12 isolates (8 NSI and 4 SI). All SI isolates used CXCR4 for entry; two used CXCR4 only, one used CXCR4, CCR5, and CCR3, and one was a mixture of two isolates using CXCR4 and CCR5. None of the NSI viruses used CXCR4 for entry. All used CCR5 as their predominant co-receptor. Of the eight NSI isolates, three used CCR5 only, two used CCR5 and CCR2b, one used CCR5 and CCR3, and one used CCR5, CCR3, and CCR2b. Log10 HIV p24 antigen production on day 14 PI for viruses that used CCR5+CCR3 (3.79 + 1.40) was greater than for viruses that used CCR5+CCR2b (3.22 + 1.55) or CCR5 (3.32 + 1.49), and all were greater than those that used CXCR4 only (1.69 + 0.28), regardless of SI phenotype (P < 0.05). Thus, in these primary isolates, macrophage tropism and replication kinetics were closely linked to CCR5 utilization, whereas SI capacity was closely linked to CXCR4 utilization. Furthermore, viruses, which could use CCR5 and CCR3 for entry, had a replication advantage in macrophages, regardless of SI phenotype.

Kernel density analysis of variable and conserved regions of the envelope proteins of human immunodeficiency virus type 1 and associated epitopes.

A new statistical approach to the study of conservation of amino acid and nucleotide sequences based on kernel density analysis is described that enables analysis of both conserved and highly variable HIV-1 protein sequences. The amino acid sequences of HIV-1 env proteins in 63 isolates were analysed to determine, first, whether the designations of regions identified in 1987 as conserved (C1-C6) or variable (V1-V5) were still valid. Even though the data base used was nine times larger, the designations that were based on seven isolates from five patients remain correct. Second, the new approach enabled the quantifications of the degree of conservation in reported B or T cell epitopes. Using this approach, highly conserved epitopes located in both gp41 and gp120 were identified.

[Cerebrospinal fluid serotonin concentration in pyogenic and tick-borne encephalomeningitis]. / Stezenie serotoniny w plynie mózgowo-rdzeniowym w przebiegu ropnych i kleszczowych zapalen opon mózgowo-rdzeniowych i mózgu.

Cerebrospinal fluid (CSF) serotonin was determined in 38 patients with pyogenic meningitis and tick-borne encephalitis (TBE) before and after treatment. Increase of CSF serotonin concentration was observed in acute phase of pyogenic meningits and normalized after treatment.

[Tuberculous meningoencephalitis from personal material]. / Gruzlicze zapalenie opon mózgowo-rdzeniowych i mózgu w materiale wiasnym.

Clinical characteristics of 6 patients with tuberculous meningoencephalitis is presented. The increased morbidity of tuberculosis in recent years makes it necessary to consider this etiology in the diagnosis of CNS infections especially with severe atypic and long lasting course. In the case when tuberculous etiology is suspected the anti tuberculous treatment should be applied. Using new and modern diagnostic methods of CSF will allow to diagnose tuberculous meningoencephalitis earlier and will improve prognosis in this disease.

[Hepatic reactions in the course of viral meningoencephalitis in patients hospitalized in 1985-1989]. / Odczyny watrobowe w przebiegu wirusowych zapalen opon mózgowo-rdzeniowych i mózgu u chorych hospitalizowanych w latach 1985-1989.

In the years 1985-1989 in a department for parasitic and occupational diseases in Bialystok, 112 patients were hospitalized with the diagnosis of lymphocytic meningitis. The most common aetiological factor were viruses (30.6%), arboviruses (25.9%), and then herpes viruses (6.25%), and mumps virus (5.3%). In 36 patients (32%) no aetiological factor of the inflammatory process was established. In 22 patients (19.6%) a simultaneous hepatocellular damage was found in the form of jaundice (15 patients, that is 13.4%), and hepatomegaly in seven patients. The observed hepatic involvement was found in the group of patients with severe clinical course of meningitis. Most frequently hepatic reaction was observed in the group of lymphocytic meningitis caused by arboviruses. Among the biochemical indices of hepatocellular damage a significant increase in relation to normal value was found of serum bilirubin level (mean = 1.75 mg%; normally mean = 1.1 mg%), alanine aminotransferase activity was 2.5 times higher than the mean normal value (mean = 77.6 IU; normal value mean = 30 IU). The observed hepatic changes were transient and regressed together with the regression of acute inflammatory state in the central nervous system.

[Changes in ECG examination of patients with trichinosis]. / Zmiany w badaniu ekg serca u chorych z wlosnica.

In the years 1963-1992, 560 patients with the diagnosis of trichinosis were treated in the Department of Parasitic Diseases and Neuroinfections, including 310 women (55.3%) and 250 men (44.7%) aged from 6 to 75 years. Out of this number of patients in 59 cases (10.5%) myocardial damage was found in the course of the disease. The most frequently found changes in ECG record were ventricular repolarization disturbances (66.1%) which persisted in 18.6% of cases before discharge from the hospital. Depolarization disturbances accounted for 32.2% of cases and persisted before discharge from the hospital in 10.1% of patients. In 6.7% of patients, persistence of pathological ECG record was found during the 4th month after the hospitalization which may be an evidence of prolongation of the inflammatory process within the myocardium.

Complete and repeatable inactivation of HIV-1 viral particles in suspension using a photo-labeled non-nucleoside reverse transcriptase inhibitor.

A method is described for achieving repeatable, complete inactivation of HIV, based on photo-inactivation of HIV reverse transcriptase (RT) with a non-nucleoside reverse transcriptase inhibitor (NNRTI), photoactive 4-[[4-[(4-azido-2,6-dimethylphenyl) amino]-2-pyrimidinyl]amino]benzonitrile (PA-DAPYa). These results show that PA-DAPYa inactivated completely a suspension of cell-free HIV-1 viral particles in a dose and time-dependent manner. Using an ELISA assay for p24, it is demonstrated that a 500nM concentration of PA-DAPYa is able to inactivate 500 TCID50 of HIV viral particles in suspension when irradiated with non-microbicidal wavelength UV light for 30min. No active p24 was detected on days 7, 14, and 21 days after culturing the inactivated HIV in peripheral blood mononuclear cells (PBMCs). Several batches of large quantities of HIV viral particles were demonstrated to be inactivated completely and repeatedly by this method. Therefore, a reliable method has been developed to inactivate HIV viral particles in a reproducible manner using an optimal concentration of PA-DAPYa and duration of UV exposure time of the treated particles. The inactivation of viral particles in suspension allows for large-scale production of an injectable formulation of inactivated HIV viral particles for vaccine development which should preserve the conformational and antigenic integrity of viral surface proteins.

Aspirin does not preferentially potentiate IgE dependent basophil CD63 upregulation in patients with food-dependent exercise-induced anaphylaxis

No disponible

Anti-idiotype-based vaccines against biological toxins.

Biological toxins produced by living organisms represent one of the major sources of contamination of stored grain and agricultural products, and other food sources. The majority of these biological toxins are highly lethal, nonproteinaceous low-molecular-weight chemical compounds which exert their potent toxicity through a variety of mechanisms. Because of their small size, they generally do not induce a significantly high affinity protective antibody response upon toxin exposure, even when conjugated to large protein carriers which enhance their immunogenicity. Moreover, the very toxic nature of biological toxins precludes their use as immunogens in the induction of protective immunity. To circumvent this difficulty, an attempt was made to develop antibody (anti-idiotype)-based vaccines against a protein synthesis inhibitor, the trichothecene mycotoxin T-2, and the sodium channel blockers tetrodotoxin and saxitoxin. Protective monoclonal antitoxin antibodies were first generated and then used to induce specific monoclonal anti-idiotype antibodies. Specific anti-idiotype antibodies were assessed for their ability to induce in vivo protective immunity against toxicity.

[Evaluation of preventive vaccination against rabies performed at the Department of Parasitic and Occupational Diseases in Bialystok in 1989-1990]. / Ocena szczepien profilaktycznych przeciw wsciekliznie przeprowadzonych w Klinice Chorób Pasozytniczych i Zawodowych w Bialymstoku w latach 1989-1990.

Between 1989-1990, 502 individuals exposed to the animals with rabies or suspected of rabies reported to out-patient clinic of agricultural occupational diseases at the K. Dluski Hospital and to the Department of Parasitic and Occupational Diseases in the Medical Academy in Bialystok. Sixty one (12.2%) of these individuals were vaccinated. Most frequently these people were attacked by dogs (70.4%), cats (11.4%), rats (6.5%), and foxes (3.2%). Rabies was diagnosed only in 9.8% of cases (Negri bodies in the brain and positive immunofluorescence test.). A vaccine inactivated on the human diploid tissue (Institute Merieux) was applied without any postvaccination complications. Transient micro-macular rash on face and chest of 3 patients, and local inflammatory reaction in 2 patients did not disturb vaccination process. Active-passive immunization was used in case of extensive lesions.

Long-term culture of human immunodeficiency virus type 1 resulting in loss of glycosylation sites.

Cultures of human immunodeficiency virus type 1 (HIV-1) provided a model for the study of mutations in the absence of host antibodies. Replicate cultures of biological and molecular clones of HIV-1 were passaged weekly for 30 or 34 weeks. Eight regions of HIV-1 genomic RNA were analyzed by means of single-strand conformation polymorphism analysis and nucleotide sequencing. Six mutations were detected in the biological clones. Two were G-->A substitutions. The frequency of mutations was higher in V1 compared to that in other regions (P = 0.01). Three mutations involved loss of potential glycosylation sites in V1. These results show that mutations in the viral genome may result from selection by factors other than host immune pressures.

[Analysis of selected parameters of damage to the liver during the course of tick-borne encephalitis]. / Analiza wybranych parametrów uszkodzenia watroby w przebiegu kleszczowego zapalenia opon mózgowo-rdzeniowych.

In 1993, 80 patients (43 female and 37 male) with tick-borne encephalitis were hospitalized in Clinic of Neuroinfections and Parasitic Diseases of Medical School in Bialystok. 6 (7.5%) patients were jaundiced, 16 (20%) had hepatomegaly. We noticed increased enzymatic activity of aspartate aminotransferase (x = 74.5 U/l) in 9 (11.25%) cases, alanine aminotransferase (x = 89.5 U/l) in 11 (13.75%) cases, increased level of bilirubin in blood (from 1.4 to 3.5 mg/dl) in 7 (8.75%) cases. Protein level in blood was average x = 5.95 g/dl, percentage of albumin was average x = 48.61%, globulin--x = 14.62%. Prothrombin index and ammonia blood level were in normal range. Observed pathological changes were transient and referred mainly to patients with severe clinical course of disease.

Single-strand conformation polymorphism study of human immunodeficiency virus type 1 RNA and DNA in plasma, peripheral blood mononuclear cells, and their virologic cultures.

Single-strand conformation polymorphism (SSCP) analysis was applied to human immunodeficiency virus type 1 (HIV-1) nucleic acids in plasma and peripheral blood mononuclear cells (PBMC) from 16 patients and to 15 PBMC cocultures and 6 plasma cultures prepared from the specimens. Two hypervariable regions were analyzed: in the gag gene and part of the V3 loop. Random paired matching of SSCP patterns between HIV-1 RNA and provirus DNA was tested, from plasma and PBMC from the same blood specimen, supernatant and PBMC from the same PBMC coculture, supernatant and PBMC from the same plasma culture, provirus DNA in cocultured PBMC and the PBMC inoculum, and HIV-1 RNA in a plasma culture supernatant and in the plasma inoculum. Paired matching was nonrandom for both regions in the first three situations and for gag in the fourth, with P < or = .01; matching was random for gag in the last situation. The HIV-1 env target region produced in culture diverged from that in the inoculum in 18 of 21 instances.

Selection of appropriate HIV-1 genomic regions for single-strand conformation polymorphism analysis of the diversity, modification, and transmission of HIV-1 quasispecies.

Single-strand conformation polymorphism (SSCP) analysis is a useful tool for studying viral quasispecies. Four regions within the HIV-1 genome were studied by means of SSCP analysis with the aim of determining which regions were the most informative for the study of HIV-1 transmission or for detection of changes in HIV-1 quasispecies populations. Nested polymerase chain reaction (PCR) was used to amplify V1, V2, V3 of the env gene, and the p2 region in the gag gene. In total, 114 plasma specimens from 79 individuals were tested, including serial specimens from 10 mother-infant pairs that were provided by the Women and Infants Transmission Study (WITS). HIV-1 in specimens that were PCR-positive with primer pair SK38/SK39 showed different percentages of positive signals with primer pairs for the four regions: V1, 63%; V2, 83%; V3, 88%, and p2, 100%. HIV-1 sequences in the p2 target region displayed the greatest degree of polymorphism. Analysis of serial specimens showed that the V1 target region was the most variable of the four regions studied and was the most appropriate region for monitoring changes in quasispecies populations. Of the four regions studied, p2 was the most informative for the study of HIV transmission, as shown by analysis of samples from documented cases of mother to infant HIV-1 transmission.