The sequencing of the complete genome of a Tomato black ring virus (TBRV) and of the RNA2 of three Grapevine chrome mosaic virus (GCMV) isolates from grapevine reveals the possible recombinant origin of GCMV.

The complete genome of a Tomato black ring virus isolate (TBRV-Mirs) (RNA1, 7,366 nt and RNA2, 4,640 nt) and the RNA2 sequences (4,437; 4,445; and 4,442 nts) of three Grapevine chrome mosaic virus isolates (GCMV-H6, -H15, and -H27) were determined. All RNAs contained a single open reading frame encoding polyproteins of 254 kDa (p1) and 149 kDa (p2) for TBRV-Mirs RNA1 and RNA2, respectively, and 146 kDa for GCMV RNA2. p1 of TBRV-Mirs showed the highest identity with TBRV-MJ (94 %), Beet ringspot virus (BRSV, 82 %), and Grapevine Anatolian ringspot virus (GARSV, 66 %), while p2 showed the highest identity with TBRV isolates MJ (89 %) and ED (85 %), followed by BRSV (65 %), GCMV (58 %), and GARSV (57 %). The amino acid identity of RNA2 sequences of four GCMV isolates (three from this study and one from GenBank) ranged from 91 to 98 %, the homing protein being the most variable. The RDP3 program predicted putative intra-species recombination events for GCMV-H6 and recognized GCMV as a putative inter-species recombinant between GARSV and TBRV. In both cases, the recombination events were at the movement protein level.

Stress response assessment of Lactobacillus sakei strains selected as potential autochthonous starter cultures by flow cytometry and nucleic acid double-staining analyses.

AIMS: The aim of this study was to apply the flow cytometry to Lactobacillus sakei strains, selected as potential autochthonous starters, to investigate dynamics and physiological heterogeneity of microbial behaviour under different stress conditions. METHODS AND RESULTS: A simultaneous nucleic acid double-staining assay was applied to discriminate cell populations in different physiological states after exposure to heat (50 and 55°C) and acid (pH 2·5 and 3·0) stresses. Alive cells with intact membranes, damaged cells still alive but with injured membranes, so with even a recovery ability, and dead cells with a permanent membrane damage were differentiated with a significant increase in damaged cells after stronger stress treatments. CONCLUSIONS: The existence and characteristics of subpopulations displaying heterogeneity in particular conditions are highly relevant, because specific subpopulations may show improved survival, changes and dynamics under stress conditions. SIGNIFICANCE AND IMPACT OF STUDY: This assay has potential for physiological research on lactic acid bacteria and for application in the food industry. The assessment of intermediate physiological states in Lb. sakei strains with recovery possibility could be an important criterion for application of potential starter cultures. Application of flow cytometry and characterization of sorted subpopulations may contribute to further understanding of diversity and heterogeneity in physiology of bacterial populations.

Isometric evaluation of rotator cuff muscles in volleyball athletes.

AIM: The aim of the present study was to evaluate, in a group of semi-professional volleyball players, the function of the rotator cuff muscles with a new specific device. METHODS: We have studied 30 asymptomatic volleyball players, 15 females and 15 males, all right handed central or lateral spikers, with some specific functional tests. Every subject was evaluated by some common clinical tests to detect shoulder range of motion (ROM) and rotator cuff muscles strength in both sides. Athletes were also submitted to an isometric bilateral shoulder strength test, with a specific upper limb dynamometer (Dynatorq), in three different positions: 1) arm 0° abducted; 2) arm 90° abducted and 90° external rotation, to try to reproduce the spike gesture; 3) arm behind the thorax, as a simulation of clinical lift-off test. All clinical and instrumental data were collected in male and female groups and, in each group, comparisons between dominant and non dominant shoulders. RESULTS: Our data show in all athletes an initial postero-superior impingement with significant weakness of the subscapularis muscle in the dominant shoulder. CONCLUSION: In all overhead athletes, and in particular in volleyball spikers, a pre-clinical diagnosis of lack of strength of the subscapularis muscle in the dominant side, before the occurrence of pain or discomfort signs, could be an important diagnostic aspect to avoid or delay the back-retraction of the joint capsule of the shoulder.

Fast Heck-Cassar-Sonogashira (HCS) Reactions in Green Solvents.

The replacement of toxic solvents with greener alternatives in Heck-Cassar-Sonogashira (HCS) cross-couplings was investigated. The fine-tuning of the HCS protocol allowed to achieve complete conversions and high speed under mild conditions. N-Hydroxyethylpyrrolidone (HEP) gave the best results. Moreover, the methodology was successfully applied to the synthesis of an intermediate of the anticancer drug Erlotinib, demonstrating the versatility of the new green protocol.

Production of antibodies to Little cherry virus 1 coat protein by DNA prime and protein boost immunization.

Little cherry, an economically important disease of cherry is caused by at least two different viruses. One of these is Little cherry virus 1 (LChV-1) for the detection of which no efficient serological tools are available, so that diagnosis is based on molecular methods. In this study, different immunization strategies for producing antibodies against the viral coat protein of LChV-1 were tried, using either purified virus preparations, or bacterially expressed protein, or a DNA vector that expressed the cloned coat protein (CP) gene in vivo. Effective induction of specific antibodies to LChV-1 CP was obtained using DNA intramuscular immunization followed by a single boost with the recombinant protein. The entire coat protein sequence was cloned in a mammalian expression vector and, after being coated by an amphiphilic non-toxic reagent was delivered into rabbit. A protein boost increased the specific immune response against the virus protein. The sensitivity of this antiserum is lower if compared with that of antisera raised conventionally against other viruses, thus it requires improvements for use for diagnostic purposes.

Synthesis of a versatile constrained analogue of dipeptide DG (Asp-Gly).

The synthesis of an orthogonally protected constrained analogue of dipeptide DG (Asp-Gly) is reported exploiting alkylation of a chiral lactam. The versatility of this analogue was proven by removal of t-Boc protecting group, followed by coupling under homogeneous conditions with t-Boc-Arg(Z(2))-Gly, to give a conformationally restricted analogue of RGDG tetrapeptide.

ERAP1 molecular characterization: Identification of a de novo allelic variant.

The novel ERAP1 allelic variant is a missense polymorphism leading to the Arg53Pro substitution.

Sodium periodate/TEMPO as a selective and efficient system for amine oxidation.

A new metal-free protocol for promoting oxidation of amines in aqueous-organic medium was developed. NaIO4 and TEMPO as the catalyst emerged as the most efficient and selective system for oxidation of differently substituted benzyl amines to the corresponding benzaldehydes without overoxidation. Unsymmetrical secondary amines underwent selective oxidation only at the benzylic position thus realising an oxidative deprotection of a benzylic group with an easy amine recovery.

Use of the spectrophotometric color method for the determination of the age of skin lesions on the pig carcass and its relationship with gene expression and histological and histochemical parameters.

The presence of lesions on the pig carcass is an indicator of poor animal welfare and has economic impact as it downgrades the carcass value. The assessment of the age of lesions on the carcass may help identify risk factors and ultimately prevent their occurrence. The aim of this study was to assess the age of lesions on pig carcasses through spectrophotometric color evaluation and to relate the results with gene expression and histological and histochemical parameters. A total of 96 barrows were mixed 4 times over 3 d before slaughter and 80 lesions were selected after skin lesion observations to define 4 age categories: < 7 h (T1), 7-25 h (T2), 25-30 h (T3), and 49-54 h (T4). A nonlesioned skin area was used as a control. At slaughter, 3 biopsies per lesion and control skin were taken immediately after bleeding for analyses of gene expression (, , , , , , , , , ), skin histological characteristics (inflammation, erosion or ulceration, and necrosis), and enzyme activity (alkaline phosphatase and adenosine triphosphatase). The number of lesions was counted on each carcass, and the color was assessed visually by a pictorial chart and instrumentally through a spectrophotometer. Delta values (Δ) were calculated as the difference between the value of the lesion and the value of the control for all measures, except for the histological analysis. Results indicated that visual color observation was not sufficiently accurate to discriminate lesions by time of infliction ( > 0.10), while the spectrophotometer ΔL* and Δa* values variation allowed the identification of < 7 h or > 25 h old lesions ( < 0.05). Similarly, the expression of , , , , and genes was higher ( < 0.05) in < 7 h old lesions, while gene expression was higher ( < 0.05) in < 25 h old lesions. As for the histological analysis, the severity of inflammation was correlated with the age of the lesion (lower score in < 7 h old lesions and higher score in > 25 h old lesions; < 0.05). To conclude, the spectrophotometric color assessment of the carcass lesions at slaughter appears to be a reliable method to discriminate between fresh and older lesions on the carcass at the abattoir.

Sentinel node biopsy after primary chemotherapy in cT2 N0/1 breast cancer patients: Long-term results of a retrospective study.

BACKGROUND: It is controversial whether sentinel node biopsy (SNB) is adequate in breast cancer patients who become cN0 after primary chemotherapy. To address this we retrospectively compared outcomes in T2 cases given primary chemotherapy, comparing those given axillary dissection (AD) with those given SNB but no AD if sentinel nodes were clinically negative post-chemotherapy. METHODS: We examined overall survival (OS), disease-free survival (DFS), and axillary failure in 317 consecutive cT2 cN0/1 patients given primary chemotherapy followed by quadrantectomy/mastectomy, between January 2002 and December 2007. The approach to the axilla changed over time allowing division into three groups: 101 (31.9%) given upfront AD; 139 (43.8%) given SNB + AD; and 77 (24.3%) given SNB only because the SNs were negative. RESULTS: After median follow-ups of 92 (AD), 99 (SNB + AD) and 72 months (SNB-only), OS (p = 0.131) and DFS (p = 0.087) did not differ between the 3 groups, or between SNB-only and the ypN1 and ypN0 subgroups of SNB + AD, or between the cN0 and cN1 subgroups (before chemotherapy) of the SNB-only group. No SNB-only patient had axillary failure. OS (p = 0.004) and DFS (p = 0.002) were better in patients with complete response than those with partial response or stable/progressive disease. CONCLUSIONS: SNB is adequate in T2 patients who are cN0 after primary chemotherapy, irrespective of axillary status before. Better outcomes after complete pathological remission confirm the prognostic importance of response to primary chemotherapy, and suggest that all T2 patients should receive primary chemotherapy.

Isolation and pathogenicity of Xylella fastidiosa associated to the olive quick decline syndrome in southern Italy.

In autumn 2013, the presence of Xylella fastidiosa, a xylem-limited Gram-negative bacterium, was detected in olive stands of an area of the Ionian coast of the Salento peninsula (Apulia, southern Italy), that were severely affected by a disease denoted olive quick decline syndrome (OQDS). Studies were carried out for determining the involvement of this bacterium in the genesis of OQDS and of the leaf scorching shown by a number of naturally infected plants other than olive. Isolation in axenic culture was attempted and assays were carried out for determining its pathogenicity to olive, oleander and myrtle-leaf milkwort. The bacterium was readily detected by quantitative polymerase chain reaction (qPCR) in all diseased olive trees sampled in different and geographically separated infection foci, and culturing of 51 isolates, each from a distinct OQDS focus, was accomplished. Needle-inoculation experiments under different environmental conditions proved that the Salentinian isolate De Donno belonging to the subspecies pauca is able to multiply and systemically invade artificially inoculated hosts, reproducing symptoms observed in the field. Bacterial colonization occurred in prick-inoculated olives of all tested cultivars. However, the severity of and timing of symptoms appearance differed with the cultivar, confirming their differential reaction.

Seroprevalence of five neglected parasitic diseases among immigrants accessing five infectious and tropical diseases units in Italy: a cross-sectional study.

OBJECTIVE: This multicentre cross-sectional study aims to estimate the prevalence of five neglected tropical diseases (Chagas disease, filariasis, schistosomiasis, strongyloidiasis and toxocariasis) among immigrants accessing health care facilities in five Italian cities (Bologna, Brescia, Florence, Rome, Verona). METHODS: Individuals underwent a different set of serological tests, according to country of origin and presence of eosinophilia. Seropositive patients were treated and further followed up. RESULTS: A total of 930 adult immigrants were enrolled: 477 men (51.3%), 445 women (47.9%), eight transgender (0.8%); median age was 37.81 years (range 18-80 years). Most of them had come from the African continent (405/930, 43.5%), the rest from East Europe, South America and Asia, and 9.6% (89/930) were diagnosed with at least one of the infections under study. Seroprevalence of each specific infection varied from 3.9% (7/180) for Chagas disease to 9.7% (11/113) for toxocariasis. Seropositive people were more likely to be 35-40 years old and male, and to come from South East Asia, sub-Saharan Africa or South America. CONCLUSIONS: The results of our study confirm that neglected tropical diseases represent a substantial health problem among immigrants and highlight the need to address this emerging public health issue.

Mannanoligosaccharides and aflatoxin B1 in feed for laying hens: effects on egg quality, aflatoxins B1 and M1 residues in eggs, and aflatoxin B1 levels in liver.

Ninety-six laying hens were allocated to 4 groups and fed diets (control diet (0-0), diet supplemented with 2.5 ppm aflatoxin B1 (0-AF); diet supplemented with 0.11% mannanoligosaccharide (MOS-0); diet supplemented with 0.11% MOS and 2.5 ppm aflatoxin B1 (MOS-AF) for 4 wk to evaluate the effect of aflatoxin B1 (AFB1), mannanoligosaccharide (MOS), or both on egg quality and the in vivo efficacy of MOS to interact with an oral administration of AFB1. After 2 and 3 wk, egg weight decreased (P < 0.05) in the group fed MOS-0 versus groups on 0-0 and 0-AF. Egg shell weight was lower (P < 0.05) in the group fed 0-AF. Aflatoxin influenced color parameters, which were probably related to interference of AFB1 with lipid metabolism and pigmentary substances deposition in yolk. MOS appeared to increase protein percentage in albumen. No AFB1 or aflatoxin M1 (AFM1; a polar metabolite of AFB1) residues were found in eggs of the experimental groups. Livers from groups 0-0 and MOS-0 always tested negative for AFB1 and AFM1. Differences (P < 0.01) were found between AFB1 hepatic levels of group 0-AF (mean +/- SD: 4.13 +/- 1.95 ppb) and group MOS-AF (mean +/- SD: 2.21 +/- 1.37 ppb). The data demonstrated the ability of MOS to adsorb and degrade AFB1, reducing gastrointestinal absorption of AFB1 and its levels in tissues.

Growth parameters, behavior, and meat and ham quality of heavy pigs subjected to photoperiods of different duration.

To attain a good level of animal welfare, pigs require a sufficient environmental illumination. Therefore, minimum levels for light duration and light intensity have been set up by the European legislation (Directive 2008/120). An experimental trial was designed to determine whether an increased duration of the photophase (up to 16 h of light per day) could modify the behavior, productive parameters, and meat and ham quality of Italian heavy pigs. Forty crossbred (Large White × Landrace) castrated males pigs (26 kg initial average BW) intended for Protected Designation of Origin (according to European Union Regulation 1151/2012, ) dry-cured ham production were raised according to Parma ham production rules up to the weight of 160 kg. Pigs were homogeneously allotted to 2 experimental groups, each comprising 20 pigs. The short photoperiod (SP) group received the minimum mandatory number of hours of light per day (corresponding to 8 h/d), whereas the long photoperiod (LP) group was subjected 16 h of light per day during the whole production cycle. Light intensity was maintained at 40 lux (i.e., the minimum mandatory level) for both the experimental groups. Growth and slaughtering parameters, carcass traits, fatty acid composition, meat and dry-cured ham quality, and animal behavior were assessed. Pigs in the LP group showed a greater live weight and carcass weight compared to the SP group ( = 0.005 and = 0.007, respectively). Similarly, hams obtained from the LP group were significantly heavier and their weight losses during the dry-curing period were reduced ( < 0.01) when compared to the SP group. No significant differences were detected between the experimental groups as concerns meat and ham quality or fatty acid composition of the subcutaneous fat. Pigs in the LP group spent more time resting and less time pseudo-rooting ( < 0.01). Our results indicate that, given an appropriate dark period for animal rest, an increased duration of the photoperiod, even at the lower mandatory light intensity level, can favorably affect growth parameters of heavy pigs without any negative effect on animal behavior, carcass traits, or meat or long-cured ham quality. Therefore, rearing pigs in semidarkness should be considered as a baseless practice, contrary to animal welfare.

Surgical resection plus tamoxifen as treatment of breast cancer in elderly patients: a retrospective study.

Between March 1982 and May 1989, 151 women with primary breast cancer ranging in age from 70 to 91 years (median 79), were treated with conservative surgical procedure followed by adjuvant tamoxifen. Surgery was performed under local anaesthesia without axillary node dissection. The median duration of follow-up was 60 months (range 36-124). There were six local, six ipsilateral axillary node and six distant relapses. Local recurrences were successfully managed with further surgery whereas axillary node relapses required radiotherapy in 3 cases, surgery in 2 cases and hormonal treatment in 1 case. 2 patients died of progression of disease and one of unrelated conditions. The 5-year relapse-free survival rate was 0.82. This treatment option in elderly patients yields an acceptable local control and reduces the risks of major surgery.

Ultrastructural modifications of the platelet plasma membrane in vascular diseases and the effect of a carbochromen derivative as studied by freeze-fracture and computer measurement.

A high surface density of the openings of the surface connected canalicular system (SCCS) has been observed in the freeze-fractured plasma membrane of circulating platelets in rabbits fed an atherogenic diet and in hypercholesterolemic type IIa patients. In vitro tests have revealed a correlation between the increased surface density of the SCCS openings and the initial steps of platelet activation. 8-Chlorocarbochromen, a new drug which enhances the in vivo release of prostacyclin from the arterial wall, has been found to be effective in reducing the high surface density of SCCS openings in platelets of rabbits on an atherogenic diet. The present study shows that circulating platelets from patients with peripheral vascular disease present a high surface density of SCCS openings compared to that observed in control subjects. After a single oral administration of 8-chlorocarbochromen, a reduction of the high number of these openings has been observed. Likewise, beta-thromboglobulin levels were found to be high in the patients and to be significantly reduced after oral administration of the drug. This study shows ultrastructural modifications of platelets in conditions related to atherosclerosis and includes data on the effectiveness of 8-chlorocarbochromen in reducing the platelet activation of these patients.

Pilomatrix carcinoma: a case report and review of the literature.

Pilomatrix carcinomas are malignant uncommon tumors of hair matrix origin. The clinicopathologic features of a case of pilomatrix carcinoma are described and compared with those of the previously reported cases. Because these tumors behave similarly to a basal cell carcinoma, a complete excision is necessary in order to minimize the likelihood of relapse.

Localized mastalgia as presenting symptom in breast cancer.

The importance of breast pain as a presenting symptom of breast cancer was assessed in 200 women with localized mastalgia but negative physical examination and in 478 women with operable breast cancer. In the first group, mammography detected five cases of subclinical breast cancer at the site of pain. In the second group, 86 patients (18%) reported localized pain as presenting symptom, followed, at different intervals, by the detection of a breast lump. Localized pain can be considered a presenting symptom of breast cancer thus requiring a careful physical and mammographic examination, especially when risk factors are associated.

Fine needle aspiration cytology in superficial lymph nodes: an analysis of 266 cases.

The results of fine needle aspiration cytology of superficial lymph nodes in 275 patients are discussed. Nine cases were lost to follow-up. Of the remaining 266 aspirates, 152 were classified as cytologically malignant, 79 as benign, three as suspicious for malignancy, and 32 as unsatisfactory owing to scanty cellularity. The sensitivity of cytology for metastatic cancer was 96.5%. The results of aspiration biopsy from lymphomas were less accurate (67.5% sensitivity). There were no false positives and nine (11.3%) false negatives. Open biopsy of three suspect lymph nodes proved them to be malignant. The aspiration procedure is easy, safe and reliable. The diagnosis of benignity cannot, however, be determined without an open biopsy, particularly for lymphomas.