B- and T-cells in the follicular fluid and peripheral blood of patients undergoing IVF/ET procedures

Problem: To analyse percentage of total and memory CD27⁺ B‐cells and other lymphocyte subpopulations in the peripheral blood (PB) and follicular fluid (FF) of infertile married couples. Method of study: Forty‐eight couples from in vitro fertilization/embryo transfer (IVF/ET) programme were divided into four groups: patients with previous unsuccessful fertilization (n = 13), ectopic pregnancy (n = 8), multiple (at least three) failed IVF/ET (n = 18) and missed abortions (n = 9). Control group consisted of 15 married couples with healthy children. Results: PB memory CD27⁺ B‐cells were significantly decreased in all groups of infertile patients compared with controls. First group had increased memory B‐cells percentages compared with the second group. The differences in the percentages of PB memory B‐cells in third and fourth group compared with the first group were not statistically significant. FF memory B‐cells in the first and third group were significantly increased compared with second and fourth group. The percentage of total FF B‐cells in all groups were significantly decreased compared with their percentage in PB. Male partners of women from the first group had had significantly increased percentages of memory B‐cells compared with the partners of women from the second group. Percentage of total T‐ and B‐cells, CD4⁺ and CD8⁺ T‐cells, NK cells and activated HLA‐DR⁺ T‐cells in all groups were not significantly different from controls. We found no statistically significant difference between immunoglobulin E levels in all groups of patients. We found lower levels of IgA and IgM in FF compared with serum in all groups. Conclusion: Infertile patients have significantly decreased percentage of CD27⁺ B‐cells in the PB. Abnormalities in the memory B‐cell compartment may contribute to the pathogenesis of infertility. In the T‐cell compartment abnormalities were not detected. It appears that hormonal stimulation did not influence cellular immunity parameters.     

Rare renal anomalies in childhood

Three patients with megacalycosis, a rare ren anomaly which includes dilatation of all ren calices, are presented. The symptoms of acute uroinfection were present in all three patients. The patients underwent clinical observation, laboratory testing, and renal ultrasound. Ultrasound revealed unilateral hydronephrosis in all three patients. Additional examinations included static and dynamic renal scintigraphy, voiding cystourethrography, and intravenous urography which pointed to unilateral megacalycosis. The symptoms of acute uroinfection were probably triggered by urinary stasis in dilated calices. Surgical intervention is not indicated in megacalycosis. The increasing incidence of uroinfection, urolithiasis and hematuria imposed the need of continuous follow-up in these patients.     

Evaluation of radioprotective effects of propolis and quercetin on human white blood cells in vitro

This in vitro study aimed at investigating the possible radioprotective effects of natural substances propolis and quercetin on gamma-irradiated human white blood cells. The levels of primary DNA damage were studied by the alkaline comet assay, while the cytogenetic damage was evaluated using the analysis of structural chromosome aberration and cytokinesis-block micronucleus assay. The results obtained by all endpoints indicate acceptable toxicity profiles of propolis and quercetin in vitro, and also confirmed their radioprotective abilities. Propolis was found to be more effective in diminishing the levels of primary and more complex cytogenetic DNA damage in gamma-irradiated white blood cells. Data gathered in present study support the use of propolis and quercetin as non-toxic protective substances. However, to clarify the underlying mechanisms of their cyto/radioprotective activities, additional studies are necessary at both in vitro and in vivo levels.     

Analysis of polychlorinated biphenyls and organochlorine pesticides in pine needles

This paper presents a review of methods for the analysis of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in pine needles. These compounds are lipophilic, they accumulate in the biotic and abiotic part of the environment in which they are persistent, and their use is forbidden or restricted due to harmful effects on human and animal health. Pine needles are convenient for monitoring the levels of organochlorine compounds in the air because they accumulate them in wax with which they are covered. However, the presence of many other compounds makes them complex for analysis. Analytical methods for determining organochlorine compounds in pine needles include sample collection, extraction of compounds with a suitable solvent, extract clean-up from unwanted compounds and qualitative and quantitative analysis. PCBs and OCPs are present in pine needles in traces, and every part of the analytical procedure has to be as efficient and selective as possible.     

Naive and memory CD4+ T-cells in the cerebrospinal fluid of children with aseptic meningitis following measles-mumps-rubella vaccination and enteroviral meningitis

We investigated the distribution of memory (CD45RO⁺) and naive (CD45RA⁺CD62L⁺) CD4⁺ T-cells as well as CD8⁺ T-cells and total T-cells in the CSF of children with aseptic meningitis following measles-mumps-rubella (MMW) vaccination and those with enteroviral meningitis. Flow cytometric analysis of CSF cells was performed in 12 children with MMR vaccine-associated meningitis and 11 children with enteroviral meningitis. Percentages of total T-cells, CD4⁺ and CD8⁺ T-cells and monocytes in CSF of patients from the two groups were not significantly different. The majority of CD4⁺ T-cells in the CSF of both patient groups were of memory phenotype. Percentages of CSF naive CD4⁺ T-cells were increased in children with aseptic meningitis following MMR vaccination. Further studies focused on the more detailed immunophenotyping of CSF cells are needed to fully establish the usefulness of flow cytometry in the diagnostic workup of inflammatory CNS diseases in children.     

The effect of poly(ethylene glycol) molecular architecture on cellular interaction and uptake of DNA complexes.

The cellular uptake of plasmid DNA complexes with a series of tertiary amine methacrylate-ethylene glycol (DMAEMA-EG) copolymers with various architectures was studied using flow cytofluorometry and laser confocal microscopy. The complexes displayed different rates and extents of cellular interaction and internalisation, depending on the copolymer molecular architecture. In general, introduction of oligo(ethylene glycol) [OEG] or poly(ethylene glycol) [PEG] chains decreased both the interaction and cellular internalisation of the DNA complexes but subtle differences were observed. Two block copolymers, a 'bottle-brush' type DMAEMA-block-OEGMA and a linear DMAEMA-block-PEG copolymer (each containing a total of 45 EG units), displayed similar uptake profiles. In contrast, only relatively low uptake of complexes formed by a comb-type statistical copolymer, DMAEMA-stat-PEGMA, was observed, despite each PEG chain comprising 45 EG units. Similar trends were observed with three cell lines, A549, HepG2 and COS-7. However, the absolute values were cell-dependent, with COS-7 cells displaying both the highest rate and extent of uptake. Studies of the association and uptake of the complexes demonstrated that cell associations generally increased over time, with the uptake level and the time profile depending on the polymer architecture. Confocal microscopy studies confirmed that, with the exception of the poorly transfecting comb-type copolymer, the association of complexes with cells resulted in endocytosis.     

Presence of carbapenem-resistant bacteria in soils affected by illegal waste dumps

The carbapenem-resistant bacteria (CRB) are currently at the top of the WHO priority list of bacteria that pose the greatest threat to human health. Considering that soil is one of the important environments for the emergence of antibiotic-resistant bacteria, we isolated and quantified cultivable CRB in soils across Croatia, including ones affected by illegal dumps.

We cultivated CRB at two temperatures, distinguishing between the intrinsically resistant CRB (37°C, mostly Stenotrophomonas spp.) and the ones that are presumably human-associated and clinically relevant (42°C, Acinetobacter sp., Enterobacteriaceae, Burkholderia spp.). Our study demonstrated that distinguishing between the two offers a better insight into the diversity of CRB in the environment. The ones cultivated at 37°C were found in almost all soil samples, while the presumably clinically relevant ones were absent from uncontaminated pasture and grassland, indicating that human-associated CRB are unlikely to be found in soils spared from anthropogenic influence.     

Immediately provisionalized tapered conical connection implants for single-tooth restorations in the maxillary esthetic zone: a 5-year prospective single-cohort multicenter analysis

Clinical Oral Investigations - This open, single-cohort, multicenter, prospective study investigated the efficacy of immediately provisionalized tapered conical connection implant for single-tooth...