Field Application of SD Bioline Malaria Ag Pf/Pan Rapid Diagnostic Test for Malaria in Greece

Greece, a malaria-free country since 1974, has experienced re-emergence of Plasmodium vivax autochthonous malaria cases in some agriculture areas over the last three years. In early 2012, an integrated control programme (MALWEST Project) was launched in order to prevent re-establishment of the disease. In the context of this project, the rapid diagnostic tests (RDT) of SD Bioline Malaria Ag Pf/Pan that detects hrp-2 and pan-LDH antigens were used. The aim of this study was to assess the field application of the RDT for the P. vivax diagnosis in comparison to light microscopy and polymerase chain reaction (PCR). A total of 955 samples were tested with all three diagnostic tools. Agreement of RDT against microscopy and PCR for the diagnosis of P. vivax was satisfactory (K value: 0.849 and 0.976, respectively). The sensitivity, specificity and positive predictive value of RDT against PCR was 95.6% (95% C.I.: 84.8-99.3), 100% (95% C.I.: 99.6-100.0) and 100% (95% CI: 91.7-100.0) respectively, while the sensitivity, specificity and positive predictive value of RDT against microscopic examination was 97.4% (95% C.I.: 86.1-99.6), 99.4% (95% C.I.: 98.6-99.8) and 86.1% (95% CI: 72.1-94.7), respectively. Our results indicate that RDT performed satisfactory in a non-endemic country and therefore is recommended for malaria diagnosis, especially in areas where health professionals lack experience on light microscopy.     

The immunologically active site of prothymosin α is located at the carboxy-terminus of the polypeptide. Evaluation of its in vitro effects in cancer patients

Prothymosin α (proTα) is a 109 amino acid long polypeptide presenting distinct immunoenhancing activity in vitro and in vivo. Recent reports suggest that in apoptotic cells, proTα is cleaved by caspases at its carboxy(C)-terminus generating potentially bioactive fragments. In this study, we identified the peptide segment of proTα presenting maximum immunomodulatory activity. Calf thymus proTα was trypsinised, and the five fragments produced (spanning residues 1–14, 21–30, 31–87, 89–102 and 103–109) were tested for their ability to stimulate healthy donor- and cancer patient-derived peripheral blood mononuclear cell (PBMC) proliferation in autologous mixed lymphocyte reaction (AMLR), natural killer and lymphokine-activated killer cell activity, intracellular production of perforin, upregulation of adhesion molecules and CD25 expression. ProTα(89–102) and proTα(103–109) significantly fortified healthy donor-lymphocytes’ immune responses to levels comparable to those induced by intact proTα. These effects were more pronounced in cancer patients, where peptides proTα(89–102) and proTα(103–109) partly, however significantly, restored the depressed AMLR and cytolytic ability of PBMC, by simulating the biological activity exerted by intact proTα. ProTα(1–14), proTα(21–30) and proTα(31–87) marginally upregulated lymphocyte activation. This is the first report showing that proTα’s immunomodulating activity can be substituted by its C-terminal peptide(s). Whether generation and externalization of such immunoactive proTα fragments occurs in vivo, needs further investigation. However, if these peptides can trigger immune responses, they may eventually be used therapeutically to improve some PBMC functions of cancer patients.     

Diet of the small-spotted catshark Scyliorhinus canicula in the Aegean Sea (eastern Mediterranean)

The diet of the small-spotted catshark Scyliorhinus canicula, captured in the Aegean Sea by bottom-trawl from 2006 to 2012, was investigated with respect to sex, maturity condition, sampling location and season. The stomach contents of 432 specimens, measuring from 144 to 517?mm in total length, were analysed. The cumulative prey curve showed that the sample size was adequate to describe the species’ diet, which was quantified using the percentage gravimetric composition (%W). The identified prey items belonged to eight major groups: Teleostei, Chondrichthyes, Crustacea, Cephalopoda, Annelida, Echinodermata, phanerogams and macroalgae, with Teleostei, Crustacea and Cephalopoda being the most consumed in both females (%W?=?48.1, 16.0 and 31.4, respectively) and males (%W?=?33.9, 31.6 and 29.8, respectively). Higher diet diversity was observed in males than females, in immature individuals than mature ones, regardless of sex, as well as in spring in comparison to autumn and winter. Feeding intensity seemed to be influenced mainly by sex and maturity condition. No significant dietary overlap was observed for all possible combinations of the factors examined. Gut indices were compared between the two sexes with females showing statistically significantly higher median relative gut length, as well as a longer gut than males of the same length. Based on the diet composition, S. canicula can be considered a generalist predator consuming, with geographical differentiation, a wide variety of benthic taxa. The estimated fractional trophic level (τ?=?4.22) classified the species as a carnivore with a preference for Teleostei and Cephalopoda, thus confirming its key role in the food web.     

Signal analysis on strings for immune-type pattern recognition

We use wavelet-type discrete transforms for signal analysis on strings of finite length. We apply these transforms for edge and hidden Markov process detection. We also present new approaches for string matching and for measures of the diversity of chaotic strings.     

West Nile Virus Seroprevalence in the Greek Population in 2013: A Nationwide Cross-Sectional Survey

Cases of West Nile Virus (WNV) disease were recorded for three consecutive years in Greece following the year 2010 outbreak. A cross-sectional serologic survey was conducted to estimate the WNV seroprevalence and assess the ratio of infection to neuroinvasive disease. A stratified left-over sampling methodology was used including age and residence strata. A total of 3,962 serum samples was collected and tested for WNV Immunoglobulin G (IgG) antibodies by Enzyme–Linked Immunosorbent Assay (ELISA). All positive samples were further tested by Plaque Reduction Neutralization Test (PRNT) and WNV Immunoglobulin M (IgM) antibodies. WNV IgG antibodies were detected in 82 samples and 61 were also positive in PRNT representing a weighted seroprevalence of 2.1% (95% C.I.: 1.7–2.6) and 1.5% (95% C.I.: 1.2–2.0), respectively. Multivariable analysis showed that seroprevalence was associated with age and residence. The overall ratio of neuroinvasive disease to infected persons was estimated at 1:376 (95% C.I.: 1:421–1:338), while the elderly people had the highest ratio. This nationwide study provided valuable data regarding the epidemiology of WNV in Greece based on the fact that elderly people have higher risk of being both infected and having severe disease.     

Thermal unfolding of human BRCA1 BRCT-domain variants

Missense mutations at the BRCT domain of human BRCA1 protein have been associated with an elevated risk for hereditary breast/ovarian cancer. They have been shown to affect the binding site and they have also been proposed to affect domain stability, severely hampering the protein's tumor suppressor function. In order to assess the impact of various such mutations upon the stability and the function of the BRCT domain, heat-induced denaturation has been employed to study the thermal unfolding of variants M1775R and R1699W, which have been linked with the disease, as well as of V1833M, which has been reported for patients with a family history. Calorimetric and circular dichroism results reveal that in pH 9.0, 5 mM borate buffer, 200 mM NaCl, analogously to wild type BRCT, all three variants undergo partial thermal unfolding to a denatured state, which retains most of the native's structural characteristics. With respect to wild-type BRCT, the mutation M1775R induces the most severe effects especially upon the thermostability, while R1699W also has a strong impact. On the other hand, the thermal unfolding of variant V1833M is only moderately affected relative to wild-type BRCT. Moreover, isothermal titration calorimetric measurements reveal that contrary to M1775R and R1699W variants, V1833M binds to BACH1 and CtIP phosphopeptides.     

Development and immunochemical evaluation of antibodies Y for the poorly immunogenic polypeptide prothymosin alpha

Since conserved mammalian polypeptides are believed to exhibit enhanced immunogenicity in avian species, hens were immunized against the poorly immunogenic, highly conserved mammalian polypeptide prothymosin alpha (ProTalpha), i.e. against either non-conjugated ProTalpha (isolated from bovine thymus) or ProTalpha conjugated to keyhole limpet hemocyanin (ProTalpha/KLH). The antibodies Y were isolated from the egg yolk and evaluated through suitable dot-blot and ELISA systems in parallel with antibodies G isolated from the antiserum of rabbits immunized against the same immunogens. As revealed, antibodies Y and G of low titer and/or affinity were obtained against non-conjugated ProTalpha, while antibodies Y against ProTalpha/KLH had a better apparent titer, could better discriminate between ProTalpha and the closely related bioactive peptide thymosin alpha 1, and were obtained at much larger quantities than the corresponding antibodies G.     

Dogfish egg case structural studies by ATR FT-IR and FT-Raman spectroscopy

The dogfish egg case is a composite structure that combines mechanical tensile strength, toughness and elasticity with high permeability to small molecules and ions. Presumably, it provides both a protective and a filtering role for the egg/embryo contained within it. In this work, we performed structural studies of the Galeus melastomus egg case at two different stages of the hardening process, utilizing ATR FT-IR and FT-Raman spectroscopy. Based on these data we deduce that: (a) The G. melastomus egg case, in close analogy to that of the related species Scyliorhinus cunicula, is a complex, composite structure which consists mainly of an analogue of collagen IV. This network forming protein appears to have common secondary structural characteristics in the entire egg case. (b) The outermost layer of the non-sclerotized egg case is especially rich in tyrosine, while the innermost layer is rich in polysaccharides, presumably glycosaminoglycans, and lipids. These differences are diminished upon hardening. (c) Disulfide bonds do not appear to play a significant role in cross-linking. However, cross-links involving tyrosine residues appear to sclerotize the egg case. It is proposed that the intensity of the Raman band at ca. 1615 cm(-1), which is due to ring stretching vibrations of Tyr, might be a useful indicator of the sclerotization status of a certain proteinaceous tissue, when tyrosines are involved in sclerotization mechanisms.     

Immunocytological and Preliminary Immunohistochemical Studies of Prothymosin ��, a Human Cancer�Cassociated Polypeptide, With a Well-characterized Polyclonal Antibody

Prothymosin α (ProTα) is a nuclear polypeptide of great biological and, possibly clinical, importance, because its expression levels have been associated with early diagnosis/prognosis of human cancer. It is therefore interesting to raise easily available and cost-effective antibodies that would be applied to develop reliable ProTα immunodiagnostics. In this study, New Zealand white rabbits and laying hens were parallel immunized against intact ProTα or the synthetic fragments ProTα[1-28], ProTα[87-109], and ProTα[101-109], all conjugated to keyhole limpet hemocyanin (KLH). The corresponding antibodies G and Y were immunochemically evaluated in parallel with ELISA and Western blot systems and applied to fluorescence immunocytology experiments using various cancer cell lines and normal cells. The antibody G raised against ProTα[101-109]/KLH had excellent functional characteristics in the Western blot and immunocytology experiments, where the fluorescent signal was almost exclusively shown in the cell nucleus independently of the cells assayed. The above antibody has been applied to preliminary IHC staining of human cancer prostate tissues, leading to a high percentage of clearly and intensively stained nuclei in the adenocarcinoma tissue; this antibody can be further used in cancer tissue immunostaining and in research concerning the role of ProTα in tumorigenesis. (J Histochem Cytochem 56:1023–1031, 2008)