Quadrivalent vaccine against human papillomaviruses (HPV)

Humani papilomavirusi (HPV) su iznimno heterogena grupa DNA virusa koja ima vrlo važnu ulogu u etiologiji različitih benignih i malignih novotvorina pločastog epitela. Prevencija HPV infekcije postiže se primjenom rekombinantnog cjepiva koje sadrži virusima-slične čestice (viral-like particles, VLP) te u cijepljenih osoba potiče sintezu neutralizirajućih protutijela specifičnih za pojedine virusne genotipove. VLPsu prazne, neinfektivne virusne kapside koje ne sadrže DNA i koje su morfološki identične nativnim virusima što ih čini odličnim kandidatima za proizvodnju profilaktičkih cjepiva. Klinička ispitivanja faze I i II dokazala su dobru podnošljivost VLP-cjepiva protiv HPV-a, njihovu sposobnost indukcije visokog titra neutralizirajućih protutijela (50–100 puta veši titar u usporedbi s prirodnom infekcijom) kao i vrlo veliku učinkovitost u zaštiti od uspostave perzistentne HPV infekcije te posljedične pojave bolesti. Kliničko ispitivanje faze III završeno je za samo jedno cjepivo protiv HPV-a tj. za četverovalentno cjepivo koje sadrži VLP genotipova HPV-6, HPV-11, HPV-16 i HPV-18 proizvođača tvrtke Merck Sharpe & Dohme (Gardasil®, Silgard®). Do siječnja 2007. g. četverovalentno cjepivo protiv HPV-a odobreno je u 52 zemlje (po žurnom postupku) uključujući SAD, 25 članica Europske Unije, Meksiko, Australiju, Novi Zeland, Peru i Brazil. Cjepivo je trenutno u postupku registracije u još približno pedesetak država. Literaturni podatci pokazuju da su tijekom dosadašnje primjene cjepiva najčešće zabilježene vrlo blage nuspojave, uglavnom lokalne reakcije na mjestu cijepljenja.Human papillomaviruses (HPV) are remarkably heterogeneous group of DNA viruses that are causally involved in the etiology of different benign and malignant neoplastic lesions of the anogenital, oropharyngeal and cutaneous epithelium. The prevention of HPV infection can be achieved by the induction of genotype-specific neutralizing antibodies triggered by recombinant L1 viral-like particle (VLP) vaccines. VLP are attractive candidates for prophylactic vaccines as they are empty, non-infectious, DNA-free viral capsids morphologically indistinguishable from the native viruses. Phase I and II trials have shown that the VLP-based HPV vaccines are well tolerated, that they induce high titers of neutralizing antibodies (50–100 fold greater than during natural infection), and protect with a very high efficacy against a persistent HPV infection and HPV-related clinical disease. Until now only one HPV vaccine completed phase III trials. The vaccine produced by Merck Sharpe & Dohme (Gardasil®, Silgard®), is a quadrivalent vaccine with HPV-6, HPV-11, HPV-16, and HPV-18 VLPs. Until January 2007, the quadrivalent HPV vaccine has been approved in 52 countries (all under accelerated review schedules) including the US, all 25 European Union member countries, Mexico, Australia, Canada, New Zealand, Peru and Brasil. Additional applications are currently under review with regulatory agencies in more than 50 countries around the world. According to the published data, the vaccine has been associated with minimal adverse reactions, the most common of which being local reactions

Difficulties in neuroborreliosis diagnostics

Neuroborelioza se može manifestirati različitim neurološkim poremećajima koje uzrokuju bakterije Borrelia burgdorferi sensu lato (s.l.). Kliničke manifestacije neuroborelioze nisu patognomonične. Etiološka dijagnoza neuroborelioze uglavnom se temelji na serološkim testovima i određivanju specifičnih protutijela za B. burgdorferi s.l. u likvoru. Fenotipske razlike vrsta B. burgdorferi s.l., razlike u njihovoj antigenoj strukturi, zemljopisnoj rasprostranjenosti te sposobnosti bolesnika za učinkovitu imunoreakciju određuju specifični humoralni odgovor. U dijelu bolesnika s neuroboreliozom protutijela se sintetiziraju samo intratekalno. Stoga je neophodno istovremeno testirati serum i likvor te odrediti indeks protutijela likvor/serum. U ovom radu analizirani su rezultati serološke dijagnostike 28 bolesnika s kliničkom dijagnozom neuroborelioze. Specifična protutijela IgM i IgG za B. burgdorferi s.l. određena su u serumu i likvoru rekombinantnim neizravnim imunoenzimskim testom (rELISA; Biomedica, Wien, Austria) te ELISA metodom »hvatanja« (engl. capture) za procjenu intratekalnih protutijela određivanjem indeksa protutijela (cELISA; IDEIALyme Neuroborreliosis, Dako, Denmark). Svi reaktivni rezultati u serumu potvrđeni su metodom Western blot (WB; Mikrogen, Germany or DPC Biermann GmbH, Germany). Specifični indeks protutijela u cELISA definiran je prema preporuci proizvođača. Intratekalna protutijela za B.burgdorferi s.l. dokazana su u 11 od 28 (39,3%) pacijenata pomoću cELISAi indeksa protutijela likvor/serum. Detektabilna protutijela u likvoru primjenom rELISA nađena su u 21 od 28 (75%) pacijenata. U 19 od 28 (67,9%) pacijenata rezultati rELISA potvrđeni su metodom WB. Sve serološke rezultate treba interpretirati u skladu s kliničkim i epidemiološkim podacima i koristiti ih za potvrdu kliničke dijagnoze, budući da serološki testovi za dokazivanje protutijela za B. burgdorferi nisu još standardizirani.Neuroborreliosis includes a variety of neurological disorders caused by Borrelia burgdorferi sensu lato. Clinical manifestation of neuroborreliosis is not pathognomonic. The etiological diagnosis is based mainly on serological tests and determination of specific anti-B. burgdorferi antibodies in cerebrospinal fluid (CSF). Specific antibodies response is influenced by phenotypic differences among B. burgdorferi species, different antigenic structure, their different geographic spreading, and patient\u27s possibility to react to infection. In some patients with neuroborreliosis antibodies could be produced only intrathecally. Therefore, it is always necessary to test serum and CSF simultaneously and determine the CSF/serum antibody index. In our study serological results of 28 patients with clinical diagnosis of neuroborreliosis were analyzed. Serum and CSF anti-B.burgdorferi IgM and IgG antibodies were determined by recombinant indirect ELISA (rELISA; Biomedica, Wien, Austria) and capture ELISA for intrathecal antibody synthesis detection by CSF/serum antibody index determination (cELISA; IDEIA Lyme Neuroborreliosis, Dako, Denmark). All sera positive results were confirmed by Western blot (WB; Mikrogen, (67,9 %) patients had confirmed rELISA results by WB. All serological results should be interpreted according to clinical and epidemiological data and used to confirm the clinical diagnosis, while serologic assays for anti-B. burgdorferi antibodies have not been standardized yet

Biological, Epidemiological and Clinical Basis of Understanding Human Immunodeficiency Virus Infection

Human immunodeficiency virus (HIV) causes a chronic infection beginning in most individuals with an acute syndrome followed by an asymptomatic stage and progresses in untreated adults over a median of 10 years to the late stage called AIDS. The virus rapidly and enormously replicates from the initiation of infection. The principal immunodeficiency caused by HIV is depletion in the subset of T lymphocytes referred to as helper T cells. New anti-HIV drugs given in potent combination regimens have demonstrated impressive efficacy by both clinical and laboratory measures, and have provided evidence that drugs can suppress HIV replication and disease manifestations. HIV/AIDS is still uncommon in Croatia. In the period from 1986 to 2000, 171 patients with AIDS have been reported of whom 101 (59%) died. The incidence of AIDS in 2000 was about 4 cases per million inhabitants. Recent testing of injection drug users at a needle exchange program (Help, Split) revealed an HIV incidence of about 1%

Immunological response in Epstein-Barr virus infection

Epstein Barrov virus (EBV) je humani herpesvirus ubikvitaran u općoj populaciji. Poput drugih herpesvirusa ima latentnu i produktivnu fazu životnog ciklusa. U EBV-infekciji razvijaju se protutijela na različite antigene. Dijagnoza i definiranje statusa EBV-infekcije mogući su zbog karakterističnog odgovora specifičnih protutijela koja se mogu određivati komercijalno dostupnim testovima. U primarnoj infekciji počinju se stvarati protutijela IgM i IgG na virusni kapsidni antigen (VCA), detektiraju se protutijela na rane antigene (EA), a izostaju protutijela IgG za EBNA. Protutijela za EBNA i VCA pokazatelj su ranije EBV-infekcije. Reaktivaciju EBV obilježava visoki titar protutijela za EA, porast protutijela za VCA i ranija prisutnost protutijela za EBNA. Kontrola replikacije EBV primarno je posredovana citotoksičnim T-limfocitima i specifičnim protutijelima za EBV-antigene. Akutnu EBV-infekciju karakterizira samoograničavajuća proliferacija pomoćničkih CD4+ te poglavito citotoksičnih CD8+ T-limfocita. Citotoksični CD8+ T-limfociti uništavaju EBV-om zaražene B-limfocite i na taj način eliminiraju akutnu infekciju nakon čega slijedi doživotna latentna infekcija. Posljednjih se godina sve više istražuje EBV-specifična stanična imunost primjenom nove tehnologije MHC tetrametra. U akutnoj fazi EBV-infekcije dolazi do intenzivne proliferacije CD8+ T-limfocita specifičnih za litičke, ali i za latentne epitope ovog virusa. Određivanje EBV-specifične imunosti posebno je značajno za istraživanje patogeneze, ranu dijagnostiku te imunoterapiju post-transplantacijske limfoproliferativne bolesti.Epstein-Barr virus (EBV) is a human herpes virus that is ubiquitous in the adult population. Like all herpes viruses, EBV has latent and productive (lytic) phases in its life cycle. Infection with EBV induces the synthesis of antibodies to various virus antigens. A characteristic pattern of specific antibodies that can be determined by commercially-available assays enables the diagnosis and the definition of EBV profile in infected patients. In primary infection IgM and IgG antibodies to viral capsid antigen (VCA) start to produce; antibodies to early antigens (EA) are detectable; and antibodies to EBNA are not present. High anti-EA antibodies titre, anti-VCA titre rising and anti-EBNA antibodies predispose EBV reactivation. Cytotoxic T-cells and specific anti-EBV antibodies primarily mediate EBV replication control. Acute EBV-infection is characterized by a self-limiting proliferation of both helper-inducer CD4+ and cytotoxic- -suppressor CD8+ T-cells. The CD8+ T-cell immune response is believed to be responsible for the elimination of acute infection. Application of MHC tetramers demonstrated a massive expansion of CD8+ T-cells specific for both lytic and latent EBV proteins at the acute stage of infection. Characterisation of EBV-specific T-cells is an important experimental tool for the investigation of post-transplant lymphoproliferative disease pathogenesis and immunotherapy

Multifactorial analysis of non-malignant lymphadenopathies

Cilj istraživanja: Povećani limfni čvorovi su česta pojava u ljudi. Vjerojatnost da je limfadenopatija nemaligne etiologije iznosi 80% u dobi do 40. godine života. Cilj ovog prospektivnog istraživanja (n=110 bolesnika) bile je etiološka, citomorfološka i citoimunološka karakterizacija nemalignih limfadenopatija. Ispitanici: 110 bolesnika koji su se javili na Odjel zarazne bolesti u Osijeku zbog povećanja jednog ili više limfnih čvorova. Metode: anamnestički upitnik, klinički pregled, rutinske hematološke i biokemijske pretrage, etiološka serološka dijagnostika, UZV pregled, citopunkcija, lančane reakcije polimeraze (PCR) za detekciju EBV DNK u uzorcima periferne krvi bolesnika, te imunofenotipizacija limfocita periferne krvi na protočnom citometru. Rezultati: Čak u 57% od ukupnog broja bolesnika s nemalignom limfadenopatijom serološkim testovima je dokazana EBV infekcija dok je u 21,8% bolesnika dokazana bakterijska infekcija. Toksoplazmoza je dokazana u 9,1% bolesnika, tularemija u 3,6%, miješana infekcija u 3,6%, bolest mačjeg ogreba u 2,7%, tuberkuloza u 0,9% i citomegalovirusna infekcija (CMV) u 0,9% bolesnika. U bolesnika s EBV limfadenopatijom od 6-18 godina (58%) najčešća lokalizacija povećanog limfnog čvora bila je submandibularno (45%), a potom ispred sternokleidomastoidnog mišića (41%) uz promjer 2-3 cm (72%). Najveći broj bolesnika imao je ukupan broj leukocita unutar referentnih vrijednosti (55%), no dominirala je limfocitoza (81%) s pojavom reaktivnih limfocita (57%), sedimantacija eritrocita je bila povišena (70%), a CRP unutar referentnih vrijednosti (57%). LDH je povišena (71%), kao i AST (84%) i ALT (87%), a svi citološki nalazi punktata limfnog čvora bili su reaktivna hiperplazija limfnog čvora. U 4 bolesnika s negativnom EBV serologijom dokazana je EBV DNK u perifernoj krvi. Analiza limfocitnih subpopulacija periferne krvi bolesnika s akutnom EBV infekcijom pokazala je porast subpopulacije T-limfocita, CD8+ T- limfocita i HLA-DR+ T-limfocita uz istovremeno smanjenje postotka B-limfocita i CD4+ T-limfocita u odnosu na referentne vrijednosti. Zaključak: Suvremene molekularne i imunofenotipizacijske metode imaju svoje mjesto u razjašnjavanju etiologije nemalignih limfadenopatija.Aim: Enlargement of the lymph nodes is a frequent finding. In patients younger than 40 years, about 80 % of lymphadenopathies are non-malignant. The aim of this prospective study (n = 110 patients) was a detailed characterisation of etiology, cytomorphology and cytoimmunology of non-malignant lymphadenopaties. Methods: Anamnestic data, clinical examination, routine hematological and biochemical methods, etiological serological diagnostics, ultrasound examination, cytological punction, polymerase chain reaction (PCR) for EBV DNA in the peripheral blood, flow cytometric immunophenotyping of peripheral blood lymphocytes. Results: EBV infection was the cause of lymphadenopathy in 57 % of patients. Bacterial lymphadenopathy was determined in 21% of patients, toxoplasmosis in 9,1 %, tularemia in 3,6 %, mixed infection in 3,6 %, cat scratch disease in 2,7 %, and cytomegalovirus (CMV) infection in 0,9 % of patients. In patients with EBV lymphadenopathy between ages 6–18 years (58 %), the most frequent localization of lymph nodes was a submandibulary region (45 %) as well as super ficial sternomastoid nodes (41 %). The majority of patients had normal total leukocyte numbers (55 %) with lymphocytosis (81 %) and the presence of reactive lymphocytes (57 %). Elevated levels of liver enzymes were also observed: LDH (71 % of patients), AST (84 % of patients), ALT (87 % of patients). Reactive lymph nodes hyperplasia was revealed by cytological examination of lymph nodes in all patients. EBV DNA was detectable in the peripheral blood of 4 patients without serological evidence of acute infection. Flow cytometry analysis revealed increased percentages of T-cells, CD8+T-cells, HLA-DR+T-cells as well as decreased percentages of B-cells and CD4+ T-cells in the peripheral blood of EBV-infected patients compared with healthy controls

Frequency of HIV-1 Viral Load Monitoring of Patients Initially Successfully Treated with Combination Antiretroviral Therapy

BACKGROUND: Although considered an essential tool for monitoring the effect of combination antiretroviral treatment (CART), HIV-1 RNA (viral load, VL) testing is greatly influenced by cost and availability of resources. ----- OBJECTIVES: To examine whether HIV infected patients who were initially successfully treated with CART have less frequent monitoring of VL over time and whether CART failure and other HIV-disease and sociodemographic characteristics are associated with less frequent VL testing. ----- METHODS: The study included patients who started CART in the period 1999-2004, were older than 18 years, CART naive, had two consecutive viral load measurements of <400 copies/ml after 5 months of treatment and had continuous CART during the first 15 months. The time between two consecutive visits (days) was the outcome and associated factors were assessed using linear mixed models. ----- RESULTS: We analyzed a total of 128 patients with 1683 visits through December 2009. CART failure was observed in 31 (24%) patients. When adjusted for the follow-up time, the mean interval between two consecutive VL tests taken in patients before CART failure (155.2 days) was almost identical to the interval taken in patients who did not fail CART (155.3 days). On multivariable analysis, we found that the adjusted estimated time between visits was 150.9 days before 2003 and 177.6 in 2008/2009. A longer time between visits was observed in seafarers compared to non-seafarers; the mean difference was 30.7 days (95% CI, 14.0 to 47.4; p<0.001); and in individuals who lived more than 160 kilometers from the HIV treatment center (mean difference, 16 days, p=0.010). ----- CONCLUSIONS: Less frequent monitoring of VL became common in recent years and was not associated with failure. We identified seafarers as a population with special needs for CART monitoring and delivery

Криостимуляция околораневых и раневых тканей в лечении длительно не заживающих ран и язв конечностей

Представлен опыт применения разработанного автором метода криостимуляции околораневых и раневых тканей, его сочетания с атравматическим адгезивным растяжением тканей у больных с длительно не заживающими ранами и язвами конечностей. Определены критерии его безопасного применения и эффекты влияния на ткани. Получены положительные результаты в восстановлении опорных и других тканей конечностей.The experience of application of the original method of cryostimulation of wound tissue and its combination with atraumatic adhesive stretching of the tissue in patients with persisting wounds and ulcers of the extremities is presented. The criteria of its safe use and effects of influence on the tissue were determined. Positive results in restoration of supporting and other tissue of the extremities were obtained

Urine-based testing for Chlamydia trachomatis among young adults in a population-based survey in Croatia: Feasibility and prevalence

<p>Abstract</p> <p>Background</p> <p>We assessed the feasibility of collecting urine samples for testing on genital <it>Chlamydia trachomatis </it>infection in a population-based survey, and prevalence of this infection among young people aged 18-25 in Croatia. In Croatia, as in the other countries of Eastern Europe, there is a lack of data on prevalence of <it>C trachomatis </it>in the general population, including young adults.</p> <p>Methods</p> <p>We sampled participants using a nationally representative, multi-stage stratified probability sample of young men and women. Detection of <it>C trachomatis </it>DNA in urine samples was performed by using a real-time PCR assay COBAS^®TaqMan^®CT Test, v2.0.</p> <p>Results</p> <p>Overall, 1005 young adults participated in the behavioural part of the survey, and 27.9% men and 37.5% women who were sexually experienced agreed to provide urine samples for testing on <it>C trachomatis</it>. Using multivariate analysis, women were significantly more likely to provide urine samples than men (aOR = 1.53, 95% CI 1.14-2.06) as were those who reported no condom use at last intercourse (aOR = 1.95, 95% CI 1.44-2.62). Prevalence of <it>C trachomatis </it>infection among those who were sexually experienced was 7.3% in men and 5.3% in women.</p> <p>Conclusions</p> <p>Population-based surveys that use probabilistic sampling are a feasible way to obtain population estimates of <it>C trachomatis </it>prevalence among young adults in Croatia, but it is challenging to obtain an adequate response rate. The prevalence of <it>C trachomatis </it>among young adults in Croatia found in this study was higher than that found in other European countries with similar survey response rates.</p