First Report of Onion yellow dwarf virus, Garlic common latent virus and Shallot latent virus on Garlic in Poland.

Garlic (Allium sativum L.) is vegetatively propagated and can be affected by a virus complex (1) consisting of two potyviruses, Onion yellow dwarf virus (OYDV) and Leek yellow stripe virus (LYSV), and two carlaviruses, Garlic common latent virus (GCLV) and Shallot latent virus (SLV) (2). OYDV, GCLV, and SLV are economically important viral pathogens of bulb garlic crops in many garlic-growing areas of the world. A general mosaic and yellowing of leaves of four garlic cultivars (Blanko, Harnas, Jarus, and Mega) was observed in 11 garlic-production fields in the Lodz, Mazowieckie, Malopolska, and Pomorskie regions of Poland in July 2012. ELISA was carried out with extracts from 29 collected garlic leaf samples to detect OYDV, GCLV, and SLV using commercial antiserum (DSMZ, Braunschweig, Germany). Results indicated that 6 samples (20.7%) were infected with OYDV, 25 samples (86.2%) were infected with GCLV, and 23 samples (79.3%) were infected with SLV. The presence of these viruses in garlic leaf samples was confirmed by reverse transcription (RT)-PCR using total RNA extracted using the Spectrum Plant Total RNA kit (Sigma-Aldrich, Munich, Germany) and primers, designed in this study, specific to the whole coat protein gene of OYDV (OYDVF 5'-TAGGGTTGGATTATGATTTCTCGA-3' and OYDVR 5'-TAGTGGTACACCACATTTCGT-3'), GCLV (GCLVF 5'-TTATAGGGACGGCACAAAATCAATCA-3' and GCLVR 5'-AATAGCACTCCTAGAACAACCATT-3') and SLV (SLVF 5'-AATYATTTACAATCGTCCAGCTA-3' and SLVR 5'-ATAATATCAATCAAATMCACACAATT-3'). Amplicons of the expected size were obtained for each virus. The amplified products were purified and sequenced in both directions. Sequence information of the CP genes of 9 OYDV, 12 GCLV, and 7 SLV isolates has been submitted to NCBI-GenBank with accession numbers KF862683 to KF862710. Sequence analysis showed that the coat protein gene of OYDV shared 86% identity with the coat protein gene of OYDV isolate MS/SW1 from Australia (GenBank Accession No. HQ258894). Comparison of the coat protein gene sequences of Polish GCLV isolates with those available in GenBank showed 85 to 91% sequence identities. Multiple sequence alignment revealed 84% nucleotide identity between the Polish isolate of SLV and an SLV isolate from Chinese garlic (AF314147) formerly referred to as Garlic latent virus (3). To the best of our knowledge, this is the first report of OYDV, GCLV, and SLV in garlic plants in Poland. The accurate identification of viruses present in garlic plants will help to use the appropriate strategies to reduce viral incidence in garlic-growing areas. References: (1) J. Chen et al. Arch Virol 146:1841, 2001. (2) A. M. G. King et al. Virus Taxonomy: Ninth Report of the International Committee on Taxonomy of Viruses. Elsevier Academic Press, San Diego, CA, 2011. (3) T. Tsuneyoshi et al. Arch. Virol. 143:1093, 1998.

First Report of Blueberry scorch virus in Elderberry in Poland.

Blueberry scorch virus (BlScV) is a member of the genus Carlavirus and one of the most widespread pathogens of highbush blueberry (Vaccinium corymbosum L.). The virus was first reported in the United States and has been reported in several countries in Europe, including Italy, Germany, the Netherlands, and Poland. Symptoms of scorch disease in highbush blueberry include necrosis of flower blossoms and leaves, shoot blight, and chlorosis. Sometimes BlScV infection is symptomless or limited to single blossoms and shoots, but all highbush blueberry cultivars are susceptible to virus infection. Cranberry (V. macrocarpon L.) and wild black huckleberry (V. membranaceum L.) are known as natural and symptomless hosts of BlScV (1). In June 2012, during the research concerning the occurrence of BlScV in plants outside Vaccinium sp., 15 leaf samples from five elderberry bushes (Sambucus nigra L., family Adoxaceae) were randomly collected from the Lodzkie region in Central Poland and three were positive in double antibody sandwich (DAS)-ELISA using specific antiserum (Agdia Inc., Elkhart, IN). To confirm the presence of the virus, total nucleic acid was extracted from ELISA-positive elderberry samples according to established protocol (T. Malinowski. Proc. 4th Int. EFPP Symposium, 445, 1996) and used in one step reverse transcription PCR. Primers were developed against the published NJ-2, BC-1, and BC-2 sequences of BlScV (GenBank Accession Nos. NC_003499, AY941198, and AY941199, respectively). The forward primer, RDP_1 (5'-ATGGCACTCACATACAGAAGTCC-3'), and the reverse primer, RDP_2 (5'-TGCCTCTTCAATGCACGATGTTC-3'), were used to amplify a 420-bp fragment of the RNA-dependent RNA polymerase gene of the virus. Amplicons of expected size were obtained from three DAS-ELISA-positive samples, while no products were observed for the negative control (DAS-ELISA-negative elderberry tissues). Sequence of one selected PCR product revealed 100, 88, and 87% nucleotide sequence identity and 100, 96, and 96% amino acid sequence identity with BC-2, NJ-2, and BC-1, respectively. BlScV-infected elderberry bushes were asymptomatic. As BlScV is transmitted by aphids in a non-persistent manner, infected elderberry bushes near highbush blueberry plantings may play an important role in virus spread. The potential for BlScV infection of plants outside family Ericaceae should be investigated. To the best of our knowledge, this is the first report of BlScV infecting elderberry. Reference: (1) R. R. Martin et al. Viruses 4:2831, 2012.

Molecular characterization of Polish Blueberry red ringspot virus isolate.

In this study, we determined the complete sequence of the genomic DNA of a Polish isolate of Blueberry red ringspot virus (BRRSV24) and compared it with a Czech (Darrow 5), and the US isolates of the virus and those of other Caulimoviridae family. The genomic DNA of BRRSV24 consists of 8,265 nucleotides and encodes eight open reading frames (ORFs). The sequence homologies of the eight ORFs of BRRSV24 were from 95 to 98% in respect of Darrow 5 and from 91 to 98% in respect of the US isolates at the amino acid level. This high level of amino acid sequence identity within the coding regions among the Czech, the US and Polish BRRSV isolates is suggestive of their common origin.

Feasibility studies and pre-design simulation of Warsaw's new wastewater treatment plant.

The proposed transfer of wastewater from the western part of Warsaw, across the Wisla (Vistula) River for joint treatment at the existing eastern side "Czajka" wastewater treatment plant (WWTP) will result in combined winter flows of approx. 580,000 m3 d(-1). One-year of pilot-scale studies defined the COD characteristics and kinetics of nitrogen removal and VFA production from primary sludge. BioWin simulation was used to size and price the optional processes and pointed to the Westbank process as the most cost-effective. The process consists of a sequence of a RAS pre-denitrification zone followed by an anaerobic, anoxic and aerobic zone. Some 100-150 t d(-1) of 10% methanol would be needed to remove 2-4 mg l(-1) of NO3-N above the recommended effluent level TN = 10 mg l(-1). Applying the principle of annual average 80% TN removal, and allowing for use of daily composite samples (rather than grab) could annually save the municipality over 1.5 million Euro on external carbon source.

Immunoperoxidase detection of polycyclic aromatic hydrocarbon-DNA adducts in breast tissue sections.

Although the etiology of the majority of human breast cancers is unknown, environmental carcinogens are suspected to play a role. In this study, we investigated polycyclic aromatic hydrocarbon-DNA adducts in 78 breast cancer patients and benign breast disease patients with lifetime environmental exposure to polycyclic aromatic hydrocarbon (PAH) compounds. Adducts were detected in paraffin sections by immunoperoxidase method using polyclonal antiserum and were quantitated by the image-analyzing system. A significantly higher level of adducts was found in benign breast disease as compared to cancer patients (P < .001; Mann-Whitney U test). Neither smoking nor genetic polymorphisms in glutathione S-transferase and cytochrome P450 influenced the level of adducts. This exploratory study demonstrates the usefulness of the immunoperoxidase method to detect PAH-DNA adducts in stored breast tissue and suggests further research on a larger population, including patients from both high- and low-pollution environments.

High frequency of recurrent mutations in BRCA1 and BRCA2 genes in Polish families with breast and ovarian cancer.

Germ-line mutations in BRCA1 and BRCA2 genes result in a significantly increased risk of breast and ovarian cancer. Other genes involved in an increased predisposition to breast cancer include the TP53 gene, mutated in Li-Fraumeni syndrome. To estimate the frequency of germ-line mutations in these three genes in Upper Silesia, we have analyzed 47 breast/ovarian cancer families from that region. We found five different disease predisposing mutations in 17 (36%) families. Twelve families (25.5%) carried known BRCA1 mutations (5382insC and C61G), four families (8.5%) carried novel BRCA2 mutations (9631delC and 6886delGAAAA), and one family (2%) harbored novel mutation 1095del8 in the TP53 gene, which is the largest germline deletion in coding sequence of this gene identified thus far. The 5382insC mutation in BRCA1 was found in 11 families and the 9631delC mutation in BRCA2 occurred in three families. These two mutations taken together contribute to 82% of all mutations found in this study, and 30% of the families investigated harbor one of these mutations. The very high frequency of common mutations observed in these families can only be compared to that reported for Ashkenazi Jewish, Icelandic, and Russian high-risk families. This frequency, however, may not be representative for the entire Polish population. The observed distribution of mutations will favor routine pre-screening of predisposed families using a simple and cost-effective test.

Measurement of cytogenetic endpoints in women environmentally exposed to air pollution.

The levels of sister chromatid exchanges (SCE), high-frequency cells (HFC) and chromosomal aberrations (CA) were studied in lymphocytes of Silesian women environmentally exposed to ambient air pollutants. Inhabitants of a less polluted but similarly urbanized area, in a rural region of Poland, served as controls. The study population was selected to minimize the major confounding factors influencing SCE and CA. These factors include age, gender, smoking status, and occupation. All donors were 35-46 years old non-smoking City Hall clerks. The levels of all three biomarkers were significantly higher in the exposed group than in controls as analyzed by the Mann-Whitney U-test. No correlation was found between levels of CA and SCE. Additional possible confounders, such as passive smoking, ex-smoking and X-ray chest examination did not influence the levels of biomarkers. This study builds upon our previous research in a male population but better controls for confounders. Thus, the results reveal genetic damage resulting from low-dose but chronic environmental exposure.

A molecular epidemiology study in women from Upper Silesia, Poland.

The present report is a follow-up to our previous molecular epidemiology studies on DNA damage in residents of the industrial region of Upper Silesia. The study was designed to focus on environmental exposure to airborne pollutants; other exposures or confounding factors (e.g. smoking status and age) were eliminated. A Silesian population consisting of 67 donors was compared to 72 inhabitants of a less polluted but similarly urbanized area, surrounded by a rural part of Poland. In both regions the donors were non-smoking females with similar age range, and occupation. Eight biomarkers including urinary mutagenicity and 1-hydroxypyrene, polycylic aromatic hydrocarbon PAH-DNA adducts in oral mucosa, sister chromatid exchanges (SCE), high frequency cells (HFC), chromosomal aberrations (CA), and sensitivity to bleomycin in lymphocytes as well as glutathione s-transferase (GSTM1)/cytochrome P4501A1 (CYP1A1) genotypes were evaluated in samples collected in summer and winter seasons. All the biomarkers of internal and biological doses of mutagens and their early biologic effects indicated statistically significant increases in the Silesian group when compared to the controls. Immunohistochemical quantitation of PAH-DNA adducts additionally revealed significant seasonal changes in the levels of adducts. No influence of susceptibility genotypes (GSTM1 and CYP1A1) on biomarker levels was observed.

Bleomycin sensitivity test in the exposed and reference human populations.

Sensitivity to bleomycin was investigated in lymphocytes collected from three groups of males: 30 occupationally exposed cokery workers, 38 environmentally exposed Silesian citizen and 35 rural inhabitants. The data were analyzed at both the individual and group levels. The first analysis has revealed a substantial interindividual variability in the level of generated breaks (breaks per cell, b/c). This variability was independent of the age of the donor, smoking habit and X-ray exposure as tested in the multiple regression model. The means per group for the occupationally and environmentally exposed persons were almost the same with the values of 0.674 and 0.639, respectively. These two groups differed significantly from the rural population (b/c=0.448, p<0.001 by MANOVA). The reproducibility of the assay was satisfying (p>0.49 by the Wilcoxon matched paired test) after omitting 7 out of 49 repeatedly sampled donors. Those persons exhibited extremely high b/c rates in the first sampling.

Carrier DNA affects the integration of HSV-1 TK gene in the genome of L929TK- cells.

L929TK- cells were cotransfected with DNA mixtures containing tk gene of HSV-1, plasmids carrying LTR of MoMLV or RSV and carrier DNA of salmon sperm or chromosomal DNA of recipient cells. Selection of TK+ transformants was conducted in DMEM supplemented with HAT. Plasmids carrying LTR sequences of MoMLV or RSV retroviruses showed enhancing effect on the frequency of TK+ transformation. Southern blot analysis of chromosomal DNA of TK+ transformants demonstrated in clones deriving from cotransfections of tk gene and carrier DNA of L929TK- cells multiple copies of tk gene integrated into several genomic sites of host. Single copies of tk gene integrated into different sites of host genome occurred in chromosomal DNA of TK+ clones deriving from cotransfections of tk gene and carrier DNA of salmon sperm. Cells cotransfected with tk gene and plasmids carrying LTR sequences of MoMLV or RSV formed three dimensional colonies in semisolid agar medium. No effect of carrier DNA on the morphology of TK+ transformant clones was noticed.

[Multiple insulin injections with the help of a subcutaneous catheter or metered syringe (Pen)]. / Wielokrotne wstrzykiwanie insuliny za pomoca podskórnego cewnika oraz dozownika insuliny (Pea).

Multiple, daily injections of insulin in intensified insulin therapy of diabetes mellitus has many important advantages. From the technical point of view however it requires new instruments, which could further limit the trauma of injection and facilitate the necessary manipulations. For these reasons a combination of typical insulin syringe (Novo-Noraisk Pen) and a special subcutaneous insulin catheter (Vigo-Spectramed Insuflon) was examined in a systematic way. Insuflon was inserted for 7 days. Insulin injection was made with a syringe through the head of the Insuflon. The needle of a syringe easily penetrated the membrane inside the Insuflon 4 times daily for 7 days without any damage. The acceptance of this method was high. It increased the compliance in general. The parameters of diabetes mellitus control changed positively. No side-effects of such procedure were noted.

[Statistical-clinical analysis of arterial blood pressure in the outpatient clinic population of patients with diabetes mellitus]. / Statystyczno-kliniczna analiza nadcisnienia tetniczego w poradnianej populacji chorych na cukrzyce.

The level of morbidity due to arterial blood hypertension was determined in a cohort of 2,492 diabetic patients, including 1,996 NIDDM and 496 IDDM cases treated at Warsaw Voiévodship Diabetic Out-Patient Clinic. In the same time, its correlation with several, selected clinical parameters was analysed statistically. It was found that: 1. Arterial hypertension morbidity rate in the whole cohort was 48.0% being significantly higher than that in the general population of Warsaw aged between 35 and 64 years and amounting to 37.4% for male and 30.9% for female patients. 2. Morbidity index from the present study was significantly higher in NIDDM cases (53.3%) than that for IDDM cases (26.6%). 3. In case of NIDDM the highest morbidity was seen in female patients (57.9%) whereas in male patients it has reached 48.1%. It was closely related to age and in lower degree to the existence of microangiopathic syndromes. 4. In IDDM morbidity for arterial hypertension was strongly affected by microangiopathic symptoms, duration of diabetes mellitus and age. 5. Borderline, systolic-diastolic hypertension was encountered in the majority of cases.

Mutagenic activity of radiosensitizers.

Seven radiosensitizers, six derivatives of nitroimidazole (coded P1 to P5 and one imidazole derivative--P6 were investigated for mutagenicity using 3 short-term tests: Ames test, prophage lambda induction and tryptophan reversion test. Out of seven investigated compounds five were not mutagenic. Only P1 derivative induces base pair substitutions. Another derivative of nitroimidazole: metronidazole induces base pair substitution and frameshift mutations. Its positive response in the prophage lambda induction test suggests that metronidazole provokes also epigenetic changes.

Mutagenic activity of some 9-aminoalkyl acridine derivatives on S. typhimurium.

The mutagenic potential of 9-(3'-dimethylaminopropylamino)-acridine and of its 1-nitro and 2-nitro derivatives was investigated by using histidine-requiring mutants of Salmonella typhimurium. The 9-(3'-dimethylaminopropylamino)-acridine exhibited a weak mutagenic activity only on one of the S. typhimurium tester strains, TA1537. The 1-nitro derivative induced mutations with high frequency in strains TA1537, TA1538 and TA98, whereas the 2-nitro derivative was substantially more mutagenic than the parent compound but it was much less mutagenic than the 1-nitro derivative. Pre-mutational damages made by the 1-nitro derivative were repaired by the uvrB gene-repair system, whereas those caused by the 2-nitro derivative could not be repaired by this system. Both the 1-nitro and 2-nitro derivatives induced some mutations in the base-pair substitution strain TA100 carrying a plasmid. The frequency of the his+ mutation induced both by 1-nitro and by the 2-nitro derivatives in strain TA101 lacking a nitro reductase was lower. These results emphasize the involvement of the nitro group in the interaction of acridine derivatives with the bacterial genome.