The role of interferon-gamma release assay in tuberculosis control

Tuberculosis is still one of the major global public health threats. Countries with low incidence must focus on exhausting the reservoir of future cases by preventing reactivation. Therefore, it is important to identify and effectively treat those individuals who have latent tuberculosis infection and who may develop active disease. The tuberculin skin test has been the standard for detection of immune response against M. tuberculosis since the beginning of the 20th century. The new millennium has brought advancement in the diagnosis of latent tuberculosis infection. The name of the new blood test is interferon-gamma release assay (IGRA). Croatia is a middle-incidence country with a long decreasing trend and developed tuberculosis control. To reach low incidence and finally eliminate tuberculosis, its tuberculosis programme needs a more aggressive approach that would include intensive contact investigation and treatment of persons with latent tuberculosis infection. This article discusses the current uses of IGRA and its role in tuberculosis control.     

Emotional Intimacy Among Coupled Heterosexual and Gay/Bisexual Croatian Men: Assessing the Role of Minority Stress

Emotional intimacy cuts across contexts as diverse as sexual motivation and satisfaction, psychological and physical health, and relational well-being. Although the experience of intimacy and its effects on sex life may be gender and sexual orientation-specific, the role of intimacy in personal and sexual relationships has been studied mostly among heterosexual individuals and couples. Using the minority stress framework (Meyer, 2003) to address this gap in knowledge, the present study comparatively explored levels and predictors/correlates of emotional intimacy, and its association with sexual satisfaction among coupled heterosexual and gay/bisexual men sampled online in a predominantly homonegative country (Croatia). Heterosexual participants (n = 860; M_age = 36.4, SD = 9.09) were recruited in 2011 and gay/bisexual participants (n = 250; M_age = 29.4, SD = 7.13) in 2013. Controlling for age and relationship duration, gay/bisexual men reported higher levels of emotional intimacy than heterosexual men. Suggesting that the role of emotional intimacy in sexual satisfaction is not sexual orientation-specific, the strength of the association between these two constructs was similar in both samples. However, internalized homonegativity, which was negatively associated with emotional intimacy in this study, remains a challenge to creating and maintaining intimacy in male same-sex relationships.     

Controlled release of lipase from Candida rugosa loaded into hydrogels of N-isopropylacrylamide and itaconic acid

The series of poly(N-isopropylacrylamide-co-itaconic acid) hydrogels, with lipase from Candida rugosa as a model protein, were synthesized by free radical copolymerization. The composition of hydrogels was varied by monomers ratio, crosslinking agent concentration and amounts of lipase, which was loaded by in situ polymerization. All samples were characterized regarding morphology. The investigation of hydrogel swelling properties revealed their pH and temperature sensitive character. Protein loading efficiency, release profiles and the specific activity yield of the released lipase were also investigated as a function of hydrogel composition, protein content and pH, at the physiological temperature of 37°C. Copolymers of N-isopropylacrylamide and itaconic acid presented high lipase loading efficiency. Another very important feature of these copolymers was that the protein release kinetic strongly depended on the pH value of the medium. The diffusion exponents values around 1 denoted that these hydrogel compositions could be adjusted to follow near zero-order kinetics. Namely, hydrogel formulations released low amounts of lipase at pH 2.20, but much higher released protein quantities were observed at pH 6.80 enabling these copolymers to be attractive candidates as site specific protein oral drug delivery systems.     

Pesticides and their binary combinations as P-glycoprotein inhibitors in NIH 3T3/MDR1 cells

The purpose of the present study was to assess do selected pesticides as well as their binary combinations act as inhibitors of P-glycoprotein (P-gp) activity of NIH 3T3 mouse fibroblasts stably transfected with human MDR1 gene (NIH 3T3/MDR1). As a result of P-gp inhibition, the increase of intracellular accumulation of a model P-gp substrate fluorescent calcein acetoxymethyl ester was measured. Pesticide and verapamil individual dose–response data were scaled and expressed as percent of maximum effect. Results showed that out of 14 pure pesticides tested, endosulfan, phosalone and propiconazole were nearly as potent as model inhibitor verapamil (EC₅₀ = 1.5 μM), while diazinon showed a lower potency of inhibiting P-gp transport activity (EC₅₀ = 58.4 μM). Concentrations of pesticides that produced the same inhibiting effect (isoboles) were combined binary. Results calculated using the isobole method revealed that diazinon caused synergistic effect in inhibiting P-gp transport activity in all combinations.     

Risk assessment of human exposure to pesticides in food

This review presents methods for the assessment of acute and chronic risk from pesticide residues in food. Multiple pesticide residues can often be found in food. Currently, there is no internationally accepted procedure for the assessment of cumulative exposure to multiple pesticide residues in food. Therefore, risk assessment is based on toxicological evaluation of single compounds in a food matrix. The international estimation of short-term intake model (IESTI) has been used to calculate acute intake. IESTI is based on "the worst-case scenario" and addresses the possibility that consumers sometimes eat large amounts of a food item, and such a large amount might contain residues at highest levels. However, it should take into account uneven distribution of pesticide residues in food. Chronic exposure is based on a deterministic approach, analogous to the calculation of the theoretical maximum daily intake.     

Ultrastructural identification of specialized endocytic compartments in macrophages of the thymic cortico-medullary zone and germinal centers of peripheral lymphatic organs of the rat.

In this study transmission electron microscopy was used to investigate the ultrastructural features of macrophages which are strategically positioned in the thymic cortico-medullary zone and the dark zone of germinal centers of peripheral lymphatic organs in adult Wistar rats. We show that this, morphologically distinct, type of macrophage displays the entire range of cytoplasmic inclusions, which structurally closely correspond to those of endosomal/MHC-II-enriched compartments of antigen presenting cells. The macrophages of the cortico-medullary zone and germinal centers contain numerous multivesicular bodies, as well as various kinds of cytoplasmic inclusions ranging from single to aggregated multivesicular/multilamellar bodies to large vacuoles. These multilamellar inclusions are composed of elongated, irregularly shaped cisternae, with abundance of internal membrane profiles and dense bodies. Often, cortico-medullary zone and germinal center macrophages contain the typical multilamellar bodies. Polysaccharides are detected by the thiocarbohydrazide-silver proteinate method within the dense bodies of these macrophages. The functional significance of cortico-medullary zone and germinal center macrophages is briefly discussed.     

Risk factors for the onset of vasospasm and rebleeding after spontaneous subarachnoid hemorrhage]

Retrospective study comprised 120 patients with spontaneous subarachnoid hemorrhage (SAH) and risk factors for vasospasm and rebleeding were analyzed. Rebleeding and vasospasm had the same incidence (26.7%). Vasospasm prolonged preoperative and postoperative hospital period (p > or = 0.05) and deteriorated the final treatment outcome (incidence of favorable outcome was decreased from 56.9% to 41.7%). Total rebleeding mortality rate was 62.5% and depended on the clinical state before rebleeding (p < 0.05). Risk factors for vasospasm were: poor clinical condition of a patient after the initial SAH, the period of 5-13 days after hemorrhage (p < 0.01) and aneurysm located on the anterior communicant artery (p < 0.01). Risk factors for rebleeding were: poor clinical condition of the patient after the initial SAH, the first 36 hours and the period of 8-11 days after the initial SAH, advanced age of the patient, severe arterial hypertension, vasospasm and aneurysm on the internal carotid artery.     

Fungus-like hyphochytrids associated with human disease.

We report two cases, with liver and brain abscess, respectively, where fungus-like organisms belonging to the Hyphochytriomycota were found at the site of inflammation together with Peptococcus in the first and Cysticercus cellulosae in the second case. This is the first time these groups of organisms have been reported in human material. The role of hyphochytrids in human pathology remained uncertain as they were found together with already known human pathogens.     

Effect of formalin fixation on pcr amplification of DNA isolated from healthy autopsy tissues

The aim of this study is to investigate the effects of formalin fixation on the degradation of DNA molecules in five different healthy tissues exempted during the autopsy, as well as the selection of the method that is most suitable for the DNA isolation. Heart muscle, liver, brain, lung and kidney tissue obtained from the healthy people who suddenly died from a violent death were used. The parts of tissue were fixed in 10% phosphate-buffered formalin as well as in 4% unbuffered formalin at room temperature. Morphology of tissue was studied using H&E staining. The DNA was isolated 6?h, 1–7 days (every 24?h), 10, 14, 28 days and 2 months after fixation using two different methods: extraction with phenol-chloroform-isoamyl alcohol as well as with PureLink Genomic DNA Kit. Yield and purity of the DNA samples were measured spectrophotometrically at 260?nm and 280?nm. The PCR amplifications of the glycerol-3-phosphate dehydrogenase 1 (GPD1, 150 bp), ß actin (ACTB, 262 bp) and ribosomal protein L4 (RPL4, 407 bp) genes were performed to evaluate the degree of DNA fragmentation. The RPL4 gene was amplified up to 72?h, ACTB gene up to 14 days and GPD1 gene up to 28 days from tissue fixed in phosphate-buffered formalin using phenol-chloroform-isoamylalcohol protocol for DNA isolation. Liver and kidney gave better results of PCR amplification, but statistical significance between tissues was not found. Preserving period, fixative and DNA extracting method are important factors for successful PCR amplification. The healthy tissue, fixed in phosphate-formalin up to 28 days, can be useful source in molecular studies. Tissues fixed in unbuffered formalin are suitable for molecular analysis up to 7 days.