Adjuvant effect of dendritic cells transduced with recombinant vaccinia virus expressing HPV16-E7 is inhibited by co-expression of IL12

Dendritic cells (DC) enhanced the immunogenicity of recombinant vaccinia viruses (rVV) expressing the E7 protein of HPV16, a tumor-associated antigen (TAA). Immunization with DC transduced by rVV generated from strain Praha or MVA induced better protection against the growth of transplanted TC-1 tumors in C57Bl/6 mice than did immunization with either of these rVVs administered alone by the same route. Interestingly, DC transduced with a double recombinant vaccinia virus expressing E7 protein together with the Th1-polarizing cytokine IL12, which has been shown to enhance the cellular response in several other systems, induced lower anti-tumor immunity than DC transduced with rVV expressing E7 protein alone. The inhibitory effect mediated by IL12 on immunization with rVV-infected DC was dose-dependent and was observed after immunization with DC transduced with IL12-expressing rVV even at low multiplicity.     

Psychological health of family caregivers of children admitted at birth to a NICU and healthy children: a population-based cross-sectional survey

Background  

There is little information in the research literature on how parents of children who spend time in a neonatal intensive care unit (NICU) adapt psychologically to the demands of caregiving beyond the initial hospitalization period. Our aim was to compare parents of NICU children with parents of healthy full-term children, looking specifically at the relationship between parental psychosocial health and child characteristics, as well as the relationship between important predictor variables and psychosocial health.     

Vanek's tumor (inflammatory fibroid polyp). Report of 18 cases and comparison with three cases of original Vanek's series

Eighteen cases of Vanek's tumors are presented. The patients included nine men and nine women between the ages of 45 and 93 years (mean, 66.2 years). Nine cases were clinically diagnosed as polyps of the gastric antrum, five cases as polyps of the stomach (not otherwise specified), one polyp was located in the ileum and the three remaining polyps in the small intestine (not otherwise specified). The thirteen polyps with available size information measured from 0.4 to 5 cm in the greatest diameter (mean, 2.2 cm). Immunohistochemically, the affections were positive for vimentin (18/18) and CD34 (15/18). All the cases negative for CD34 also lacked concentric onion skin-like formations of the spindle cells around glands and vessels. The different immunophenotype and absence of concentric formations could be explained by the existence of two different lesions commonly designated as Vanek's tumor (inflammatory fibroid polyp) or by the hypothesis of various evolutional stages. In the differential diagnosis, it is important to distinguish namely eosinophilic gastroenteritis, gastrointestinal stromal tumor, inflammatory pseudotumor, hemangioendothelioma, and hemangiopericytoma. In contrast to gastrointestinal stromal tumors, genetically no substitution, deletion, or insertion occurred in c-kit exon 11 in all analyzed samples. Likewise, no deletion or insertion in part of c-kit exon 9 was observed.     

Daphnia intoxicated by nerve agent tabun can be treated using human antidotes

Application of human antidotes against nerve agent intoxications to microcrustacean Daphnia magna (Crustacea, Cladocera) intoxicated by a nerve agent tabun (O-ethyl-N,N-dimethylphosphoramidocyanidate) and their efficacy was investigated. It was found that antidotes can be successfully applied to intoxicated daphnids. Three different treatment regimens were tested: the combination of atropine and acetylcholinesterase reactivator (trimedoxime was chosen), atropine only and trimedoxime alone, too. The most efficient was the combination of atropine and trimedoxime followed by treatment with atropine only. The proportion of recovered animals increased with time not only in treated groups but also in the control as well. This can be explained by a spontaneous reactivation of tabun-inhibited cholinesterase in daphnids probably indicating a difference between mammalian and crustacean cholinesterases.     

High genotypic diversity of Pseudomonas aeruginosa strains isolated from patients with cystic fibrosis in the Czech Republic

Over the last decade, epidemic and frequently multidrug-resistant Pseudomonas aeruginosa has increasingly been found among European cystic fibrosis (CF) patients. In the Czech Republic, more than half of the registered CF patients attend the Prague CF centre. At this centre, a Burkholderia cenocepacia strain was recently found to have spread among the patients. The aim of the present study was to assess whether P. aeruginosa isolates from patients at this centre were also genetically related and, if so, whether they were multidrug-resistant. We investigated a collection of 69 isolates from as many patients who represented 80% of the total number of P. aeruginosa-positive patients in 2004. The organisms were typed by AFLP and SpeI macrorestriction analyses (PFGE). Using these methods, 44 unique strains and nine groups of two to five isolates each were distinguished. Among these groups, two each had a prevalence of 7% in the patient population, while others had a prevalence of < or =3%. The diversity observed with PFGE was largely in agreement with the diversity found by AFLP analysis. All isolates were susceptible to colistin; 94-96% were susceptible to piperacillin, ceftazidime, cefepime, meropenem, amikacin or tobramycin; and 84-87% were susceptible to ciprofloxacin, gentamicin or netilmicin. In conclusion, the organisms recovered from Czech CF patients showed high genotypic diversity and good susceptibility to antipseudomonal agents. The absence of highly epidemic P. aeruginosa strains may result from infection control measures taken upon recognition of the epidemic B. cenocepacia.     

High extracellular K+ evokes changes in voltage-dependent K+ and Na+ currents and volume regulation in astrocytes

[K⁺]_e increase accompanies many pathological states in the CNS and evokes changes in astrocyte morphology and glial fibrillary acidic protein expression, leading to astrogliosis. Changes in the electrophysiological properties and volume regulation of astrocytes during the early stages of astrocytic activation were studied using the patch-clamp technique in spinal cords from 10-day-old rats after incubation in 50 mM K⁺. In complex astrocytes, incubation in high K⁺ caused depolarization, an input resistance increase, a decrease in membrane capacitance, and an increase in the current densities (CDs) of voltage-dependent K⁺ and Na⁺ currents. In passive astrocytes, the reversal potential shifted to more positive values and CDs decreased. No changes were observed in astrocyte precursors. Under hypotonic stress, astrocytes in spinal cords pre-exposed to high K⁺ revealed a decreased K⁺ accumulation around the cell membrane after a depolarizing prepulse, suggesting altered volume regulation. 3D confocal morphometry and the direct visualization of astrocytes in enhanced green fluorescent protein/glial fibrillary acidic protein mice showed a smaller degree of cell swelling in spinal cords pre-exposed to high K⁺ compared to controls. We conclude that exposure to high K⁺, an early event leading to astrogliosis, caused not only morphological changes in astrocytes but also changes in their membrane properties and cell volume regulation.     

Monoclonal antibodies reactive with myelin basic protein

New monoclonal antibodies (MAbs) to myelin basic protein (BP) reveal epitopes to be in sequences 22–34, 75–82, 83–96, 118–131 and 125–131. Comparison of these results with those previously reported suggest that almost every sequence of about 10 amino acid residues may be sufficiently antigenic to make a single MAb but that certain regions are immunodominant, strong enough to make practically the same MAb repeatedly. One of these new MAbs (clone 3) has especially interesting reactivity, sharply limited to residues 75–82 in bovine and porcine BP: Lys-Ala-Gln-His-Gly-Arg-Pro. Whales presumably have the same sequence, since their BPs are fully reactive with clone 3 MAb, but all other species of BP, with known sequences of BP, have at least two changes in this sequence. Deletion of Lys⁷⁵ (as in a tryptic peptide of porcine BP) reduces reactivity with the MAb about 10-fold, whereas substitution of Ala⁷⁶ by Ser (as in all other species of BP) and either deletion of Gln⁷⁷ (as in human, monkey and rabbit BP) or His⁷⁸ (as in the guinea pig and rat BP) or substitution of Pro⁸² by Thr (as in human, monkey, rat and mouse BP) eliminates reactivity. We speculate that woodchuck and prairie dog BPs in this region closely resemble chicken BP, which has about 2% of the original reactivity. However, squirrel BP is unique, probably having only one of the changes in this region of BP, since it possesses 10–20 times the reactivity of chicken BP but still only 20–50% of the original reactivity with clone 3 MAb, a degree of reactivity not seen with any other species of BP.     

Expression of growth-associated protein 43 in the skin nerve fibers of patients with type 2 diabetes mellitus

The growth-associated protein 43 (GAP-43) is known as a marker of regenerating nerve fibers and their continuous remodeling in the adult human skin. The purpose of this pilot study was to investigate a possible role for GAP-43 in the detection of the early stages of small-fiber neuropathy in patients with type 2 diabetes mellitus (DM2) as compared with a well- established and validated parameter - intra-epidermal nerve fiber density (IENFD) of protein gene product 9.5 (PGP 9.5) immunoreactive intra-epidermal C fibers. In a group of 21 patients with DM2 within three years of diagnosis (13 men, 8 women; mean age 53.9±12.8; range 30-74) and a group of 17 healthy volunteers (8 men, 9 women; mean age 55.8±8.5; range 45-70 years), skin punch biopsies were taken from a distal calf and double immunostained with both PGP 9.5 and GAP-43. In healthy controls, 96.8% of 629 PGP 9.5 immunoreactive fibers were immunostained with GAP-43; the proportion of PGP 9.5 intra-epidermal nerve fibers immunoreactive for GAP-43 in control subjects ranged from 86.5 to 100%. In DM2 patients, IENFD was significantly lower compared to controls (median, 1.5 vs. 11.2/mm; p<0.001). The proportion of GAP-43 immunoreactive intraepidermal nerve fibers was significantly lower in DM2 patients compared to healthy controls (73.6% of 337 PGP 9.5 positive fibers; p<0.001); ranged from 0 to 98.1%. In conclusion, these results show that impaired regeneration of intra-epidermal C fibers in the early stages of type 2 diabetes mellitus, as indicated by GAP-43, might be a marker of incipient diabetic neuropathy.