Biofilm Formation on Stainless Steel by Streptococcus thermophilus UC8547 in Milk Environments Is Mediated by the Proteinase PrtS.

In Streptococcus thermophilus, gene transfer events and loss of ancestral traits over the years contribute to its high level of adaptation to milk environments. Biofilm formation capacity, a phenotype that is lost in the majority of strains, plays a role in persistence in dairy environments, such as milk pasteurization and cheese manufacturing plants. To investigate this property, we have studied S. thermophilus UC8547, a fast-acidifying dairy starter culture selected for its high capacity to form biofilm on stainless steel under environmental conditions resembling the dairy environment. Using a dynamic flow cell apparatus, it was shown that S. thermophilus UC8547 biofilm formation on stainless steel depends on the presence of milk proteins. From this strain, which harbors the prtS gene for the cell wall protease and shows an aggregative phenotype, spontaneous mutants with impaired biofilm capacity can be isolated at high frequency. These mutants lack the PrtS expendable island, as confirmed by comparison of the genome sequence of UC8547Δ3 with that of the parent strain. The prtS island excision occurs between two 26-bp direct repeats located in the two copies of the ISSth1 flanking this genomic island. The central role of PrtS was confirmed by analyzing the derivative strain UC8547Δ16, whose prtS gene was interrupted by an insertional mutation, thereby making it incapable of biofilm formation. PrtS, acting as a binding substance between the milk proteins adhered to stainless steel and S. thermophilus cell envelopes, mediates biofilm formation in dairy environments. This feature provides S. thermophilus with an ecological benefit for its survival and persistence in this environment.IMPORTANCE The increased persistence of S. thermophilus biofilm has consequences in the dairy environment: if, on the one hand, the release of this microorganism from biofilm can promote the fermentation of artisanal cheeses, under industrial conditions it may lead to undesirable contamination of dairy products. The study of the molecular mechanism driving S. thermophilus biofilm formation provides increased knowledge on how an ancestral trait affects relevant phenotypes, such as persistence in the environment and efficiency of growth in milk. This study provides insight into the genetic factors affecting biofilm formation at dairy plants.

High Varroa mite abundance influences chemical profiles of worker bees and mite-host preferences.

Honeybee disappearance is one of the major environmental and economic challenges this century has to face. The ecto-parasitic mite Varroa destructor represents one of the main causes of the worldwide beehive losses. Although halting mite transmission among beehives is of primary importance to save honeybee colonies from further decline, the natural route used by mites to abandon a collapsing colony has not been extensively investigated so far. Here, we explored whether, with increasing mite abundance within the colony, mites change their behaviour to maximize the chances of leaving a highly infested colony. We show that, at low mite abundance, mites remain within the colony and promote their reproduction by riding nurses that they distinguish from foragers by different chemical cuticular signatures. When mite abundance increases, the chemical profile of nurses and foragers tends to overlap, promoting mite departure from exploited colonies by riding pollen foragers.

Omalizumab treatment in patient with severe asthma and Eosinophilic Granulomatosis with Polyangiitis . A case report.

Eosinophilic Granulomatosis with Polyangiitis (EGPA), formerly named Churg Strauss Syndrome, is a multisystem disorder characterized by chronic rhinosinusitis, asthma, and prominent peripheral blood eosinophilia; it is classified as a vasculitis of the small and medium sized arteries, although the vasculitis is often not clinically apparent in the initial phases of the disease. We present the case of a woman with EGPA who was frequently treated with high dose steroid therapy during hospital admission for refractory asthma. After December 2008, the date when we started Omalizumab, we observed a significative reduction of circulating eosinophils and IgE serum level, and the patient was no more hospitalized for respiratory failure or asthma attacks.

Terlipressin infusion induces ischemia of breast skin in a cirrothic patient with hepatorenal syndrome.

We report a case of a 65-year-old woman with hepatitis C virus-related decompensated cirrhosis with hepatorenal syndrome, treated by high dose of terlipressin. Few hours after the highest dose was started, the patient complained burning pain in breasts, followed by the development of extensive bilateral cyanosis of breast's skin. When terlipressin was immediately stopped, pain and skin cyanosis rapidly disappeared. The peculiarity of our case is that cyanosis did not develop in common peripheral sites (e.g. fingers, toes, etc.) but in an atypical area, as skin of the breasts. Probably, this particular behaviour could be explained by the anatomical position of her large size breasts, that resulting as an extremely sloping and stretching region thus filling the maximum effect of gravity.

Subtle effect of Xenos vesparum (Xenidae, Strepsiptera) on the reproductive apparatus of its male host: Parasite or parasitoid?

Parasitic castration is an adaptive strategy where parasites usurp the hosts' reproductive physiology to complete their life cycle. The alterations in the host traits vary in their magnitude, from subtle changes in the host morpho-physiology and behaviour to the production of complex aberrant phenotypes, which often depend on the host gender. The strepsipteran macroparasite Xenos vesparum induces dramatic behavioural and physiological changes in its female host, the paper wasp Polistes dominula, while its effect on the male phenotype is largely unknown. In this study we investigated how a single X. vesparum parasite influences the functional morphology of P. dominula male reproductive apparatus. We performed morphometry and ultrastructure characterization of corpora allata, testes, seminal vesicles and accessory glands in parasitized and unparasitized males, and also in young and old males to control for the effect of age on the natural deterioration of these organs. Our results show that age significantly affects the development of male reproductive apparatus. A low parasite load - one parasite per host is the common prevalence in the field - has only a marginal impact on the reproductive morphology of P. dominula males, affecting quantitatively but not qualitatively the protein content of male accessory glands. Thus, in male P. dominula wasps, X. vesparum appears to behave as a true "parasite", in clear opposition to the role of "parasitoid" that it takes in female hosts where castration causes the reproductive death.

Effects of the combined treatment with thalidomide, megestrol and interleukine-2 in cirrhotic patients with advanced hepatocellular carcinoma. A pilot study.

BACKGROUND: Thalidomide, an anti-angiogenic agent, does not have a good therapeutic effect for advanced hepatocellular carcinoma when used alone. Megestrol and interleukin-2 have been proposed as a palliative treatment for hepatocellular carcinoma. AIMS.: We assessed the effectiveness/safety of a combined therapy with thalidomide+megestrol+interleukin-2 in cirrhotic patients with advanced hepatocellular carcinoma. PATIENTS AND METHODS: Nine cirrhotic patients with advanced hepatocellular carcinoma received oral megestrol (160 mg/day) and thalidomide (from 50 mg/day to the maximal tolerated dose). Four patients also received subcutaneous interleukin-2 (1 million U/day for 21 days/month). RESULTS: The maximal tolerated dose of thalidomide was 150 mg/day. All patients complained of sedation and other neurological or digestive adverse effects. In all but one patient the adverse effects disappeared after thalidomide withdrawal or dose reduction. Interleukin-2 administration caused a flu-like syndrome and a reaction at the injection site. During treatment, alpha-fetoprotein increased in six patients, remained stable in two and decreased in one. Eight patients showed tumour progression and one had a stable disease. Eight patients died. The median survival was 9.9 (range 2.6-18.6) months. CONCLUSION: In cirrhotic patients, the combined treatment with thalidomide+megestrol (+/-interleukin-2) does not control hepatocellular carcinoma growth, possibly due to the low tolerance to thalidomide and interleukin-2 preventing the use of appropriate dosages.

Identification of fungi from dairy products by means of 18S rRNA analysis.

The role of fungi as cause of spoilage of dairy products, such as cheese and yoghurt, has been clearly demonstrated. Despite of this, there is still a lack in rapid methods for the identification of food-associated fungi. In the course of the present work, molecular taxonomical techniques were developed and used to identify yeasts involved in the spoilage of yoghurt and moulds responsible for spoilage of vacuum-packaged hard cheese. Three methods for DNA extraction and purification were evaluated and the fungus-specific primers TR1 and TR2 were used to amplify a 581-bp fragment within the gene, coding for the small ribosomal subunit (18S rRNA) of fungi. The 18S rRNA sequence analysis of fungi isolated from yoghurt and packaged cheese allowed to identify yeast belonging to Zygosaccharomyces microellipsoides and moulds belonging to Penicillium chrysogenum and Cladosporium cladosporoides.

Indigenous raw milk microbiota influences the bacterial development in traditional cheese from an alpine natural park.

Nostrano di Primiero is a 6-month ripened cheese produced from raw milk collected in the Paneveggio-Pale di San Martino Natural Park area in the Italian Dolomites. In summer, this cheese is made using milk collected from two different areas, Passo Rolle and Vanoi, in the Paneveggio Natural Park. During the experiment, the milk from the two areas was separately processed, and cheeses were made in the same cheese factory using the same technological process. The microbiota of raw milk and cheeses of the two areas was isolated and the dominant population was monitored by RAPD analysis and identified by 16S rRNA sequence. The milk of the Passo Rolle area was mainly composed of mesophilic strains, thermophilic Streptococcus thermophilus, and low amounts of enterococci were also found; the milk of the Vanoi area was dominated by mesophilic microbiota mostly Lactococcus lactis ssp. cremoris and ssp. lactis and Lactobacillus paracasei ssp. paracasei. The plating of the natural starter culture revealed the presence of a relevant community of thermophilic cocci and lower amounts of enterococci. The dynamic population analysis showed the importance of the natural starter culture in the first 2 days of cheese ripening in both cheeses. Moreover, the large biodiversity observed in the raw milks was also detected in the cheeses during ripening. The Vanoi cheese was dominated by Enterococcus faecium and Streptococcus macedonicus in the first two days and mesophilic 21 Lb. paracasei ssp. paracasei became the most represented population after 15 days of ripening. In the first few days, the Rolle cheese was characterized by being mainly composed of thermophilic S. macedonicus and S. thermophilus and secondarily by mesophilic cocci. During ripening, the microbiota composition changed, and at 15 days, mesophilic lactobacilli were the dominant population, but later, this was mainly composed of mesophilic cocci and lactobacilli. The taxonomical identification by 16S rRNA sequence confirmed a large biodiversity related to raw milk microbiota and only five strains of S. macedonicus, Lactobacillus plantarum, 21 Lb. paracasei ssp. paracasei, Lactobacillus fermentum and E. faecium were detected in both cheeses.

Molecular characterization of the Lactobacillus community in traditional processing of Mozzarella cheese.

The natural Lactobacillus community involved in traditional Mozzarella cheese production has been investigated. The bacterial associations of whey, curd before stretching and Mozzarella were analyzed using randomly amplified polymorphic DNA (RAPD) to follow growth kinetics, and 16S rDNA sequencing to identify the taxonomical position of isolated strains. Analysis of RAPD fingerprints revealed that the Lactobacillus community was composed of 13 different biotypes and the sequence analysis of 16S rDNA demonstrated that the isolated strains belong to L. plantarum, L. fermentum, L. helveticus and L. casei subsp. casei. In addition, two strains of Weissella hellenica were isolated on selective media for lactobacilli. The four L. casei subsp. casei strains and W. hellenica contained sequences related to the prtP gene coding for proteinase, and the highest proteolytic activity in milk was found in one strain of L. casei subsp.casei.

Outgrowth of interleukin-2-dependent T cell lines with cytotoxic activity against mesothelial cells from cultures of peritoneal effluent cells of children on CPD.

Previous studies on the peritoneal immune system described the presence of activated T lymphocytes in peritoneal effluents (PE) from patients on chronic peritoneal dialysis (CPD), and showed that mesothelial cells (MC) can present antigens to T cells. In order to better define phenotypic and functional characteristics of T cells and their interactions with MC, we isolated PE cells from 15 children. At the immunophenotypic analysis, high percentages of activated T cells were identified (mean value: 15% double staining for CD3/DR; 12% CD25+). T cells with gamma/delta T cell receptor (mean 5%) and natural killer cells (mean 17%) were also present in elevated numbers. MC lines (n = 7) and interleukin-2-dependent T cell lines (9 CD4+; 1 CD8+) were also obtained by incubating PE cells under different conditions. Two cell lines showed a major histocompatibility complex (MHC) restricted cytotoxic activity against autologous MC; two lines killed allogeneic MC; one line killed both autologous and allogeneic MC. Although the hypothesis that activated T cells could kill MC after recognition of surface structures modified by dialysis fluid, or during antigen presentation, needs to be further investigated, our data suggest that the subsets of lymphocytes we identified could play an important role in the mechanisms of peritoneal membrane defense.

Peritoneal mesothelial cells produce complement factors and express CD59 that inhibits C5b-9-mediated cell lysis.

The CD59 membrane protein confers protection from C5b-9-mediated cell lysis. Because evidence exists for complement (C) activation and generation of C5b-9 in the peritoneal cavity during chronic peritoneal dialysis (CPD), we investigated, on mesothelial cell (MC) lines, the expression of CD59 and the production of C components. Four MC lines were obtained from children on CPD, and two from non uremic children. CD59 expression on MCs was investigated with anti-CD59 monoclonal antibody (mAb) and polyclonal goat immunoglobulin G (IgG). MC lines were positive for staining with anti-CD59 mAb. Western blotting analysis of MC membrane demonstrated a band with the same molecular weight as CD59. Incubation of MC with anti-CD59 mAb abrogated the protective effect of CD59 (100% cytotoxicity). C3, C4, and C6 were detected in the supernatants of MC; in non uremic MC supernatants, C5, C7, C8, and C9 were also detectable, and C4 concentration was tenfold higher. CD59 expression confers to MCs protection from C5b-9-mediated lysis. MCs produce C factors. These findings suggest that production of complement components and expression of CD59 on MCs could play a role both in peritoneal cavity infection (decreased complement production) and in peritoneal membrane damage (decreased CD59 expression and reduced remesothelialization owing to MC lysis).

Ultrastructure of fetal human gonad before sexual differentiation and during early testicular and ovarian development.

The development of the human gonad was investigated by light and transmission electron microscopy in 20 embryos and fetuses between 4.5 and 11.5 weeks of gestation, i.e. during the stages of sex-indifferent gonad, initial testicular and ovarian development. The gonadal blastema in 4.5-week-old embryos appeared formed by poorly differentiated somatic mesothelial cells, and by specialized germ cells (PGCs) with signs of ameboidism, cellular structures suggesting active protein biosynthesis and mitotic activity. The sexual differentiation of the gonads was clearly observed in 7-week-old embryos and involved at the same time the testis and the ovary. The former contained seminiferous cords formed by palisades of poorly differentiated Sertoli cells, which were segregated from the mesothelium by a rudimentary albuginea. The interstitial tissue at this age contained mesenchymal cells. Between 8 and 11.5 weeks of gestation, there was a synchronous cytodifferentiation of both Sertoli and Leydig cells. The latter acquired features of steroidogenic elements. The ovaries of 7-week-old fetuses contained packed ovigerous cords formed by somatic and germ cells (oogonia). The former embraced the oogonia with thin overlapping cytoplasm projections, and acquired features similar to those of cells in primary follicles, already at this early fetal age. At the same time the sexual differentiation of the gonads involved somatic and germ cells. In the female, the oogonia continued to show the main features they had during migration and colonization, including a high mitotic rate, signs of ameboidism and a developed apparatus for protein synthesis. On the contrary Gonocytes enclosed in the seminiferous cords progressively entered a quiescent phase characterized by a reduced mitotic rate, a decrease of endoplasmic reticulum and nucleolar complexity. The chronological relationship between the cytodifferentiation of Sertoli and Leydig cells, and changes of germ cells, suggest that somatic components of the testis may contribute to a male type of differentiation of germ cells from the very beginning of sexual differentiation.

[Treatment of premenstrual syndrome. Experience and considerations]. / Trattamento della sindrome premestruale. Esperienza e considerazioni.

The Authors review the scientific literature on premenstrual syndrome (PMS), focusing on the diagnostic standards and the complex symptomatologic pattern. They report, furthermore, the results of a study of 4 groups of patients affected by PMS and treated with different therapeutic programs. The data obtained confirm that treatment of PMS must be personal and valued on the ground of dominant group of symptoms.

[Spontaneous abortion. Epidemiologic considerations]. / Aborto spontaneo. Considerazioni epidemiologiche.

The authors report on the incidence of spontaneous abortion, the commonest complication of the pregnancy. The period examined is from March 1989 and March 1991, in the obstetrical division of University of L'Aquila. The incidence and association between maternal age, malformations, parity, maternal cigarette smoking and alcohol are reported.

[Cervico-vaginal screening in the Abruzzo region]. / Lo screening cervico-vaginale nella regione Abruzzo.

This study focuses on cervical cancer and the importance of establishing a suitable screening programme. The results of a study carried out in the Abruzzo region during a 10-year sample period are reported. A Population of 36,663 patients, of which 3,372 had never been screened, was included in the study. All patients were attending the Centre of Cytology and Colposcopy at the Clinic of Obstetrics and Gynecology in L'Aquila. In order to make progress in both the diagnosis and treatment of this pathology, it is necessary not only to obtain an interdisciplinary approach with constant cooperation between professional colleagues (cytologist, colposcopist and histologist), but also an increased sensitivity of the population to the need for screening.

[Identification of growth hormone in human embryos and fetuses]. / Individuazione del growth hormone in embrioni e feti umani.

The authors report a study on 10 human fetuses aged between 9 and 16 weeks of gestation. A PAP technique (peroxidase-anti-peroxidase) was used to identify the time and place of growth hormone appearance. Data obtained show intense positivity for chondrocytes, obleoblasts and perichrondrium. Hepatocytes were also found to be positive. These results suggest that GH plays an important role in a number of tissues in which it conditions maturation and development.

[Indications and contraindications for normal labor in patients who previously underwent cesarean section]. / Indicazioni e condizioni permittenti al travaglio di parto in pazienti già cesarizzate. 10 anni di esperienza.

In line with previously published data from all over the world, the Authors are in favour of vaginal delivery as a valid and safe alternative in women who have had a previous cesarian section, provided they are selected following appropriate screening protocol, as described in this paper. A retrospective study was performed to analyse data over a 10-year period in the Department of Ostetrics and Gynecology of the University of Aquila: of 1253 selected patients, 674 (54%) had a normal vaginal delivery, whereas 577 (46%) had a repeat cesarian section following the onset of fetal distress; no case of uterine scar rupture was recorded.

[Vulvar lesions caused by Papillomavirus (HPV): effectiveness of thymopentin (Syntomoduline). First results and experiences]. / Lesioni vulvari da papillomavirus (HPV): efficacia della timopentina (Sintomodulina). Nostri primi risultati ed esperienze.

The study included two different sized groups of patients suffering from various degrees of valvular microcondylomatosis. A group of 40 patients was treated using combined therapy while a second group of 20 patients received immunomodulating therapy alone with Sintomodulin. In particular, possible changes in the T-cell line of the immune system and correlations between the viral lesion and variations in serum levels of lymphocyte subpopulations were assessed. Results were analysed up to one year after the end of therapy.

[Continuous peridural analgesia-anesthesia during labor. Experiences and considerations]. / Analgo-anestesia peridurale continua in travaglio di parto. Nostre esperienze e considerazioni.

Fifty pregnant women attending the Obstetrics Department of the University of Aquila were included in the study. Patients were matched for age, weight, height, parity, Bishop index, temperature, arterial pressure, and cardiotopography. Twenty-five patients were assigned to the control group, the remaining twenty-five received peridural analgo-anesthesia during labour and birth so as to create a continuous lumbar epidural block. The results confirmed that this method of anesthesia is an aid against psycho-physical stress and pain in pregnant women.

[Validity of psychoprophylaxis in obstetrics. Authors' experience]. / Validità della psicoprofilassi ostetrica. Nostra esperienza.

The Authors report the results based on 20 years of practice on obstetric psycho-prophylaxis (PPO). Data on presence at course, on frequency, on primipares/pluripares ratio, on labour, on timing and mode of delivery, are assembled. Moreover, neonatal status at birth and at 10th day of life, are investigated. The data obtained were compared with a control group, constituted by women without any treatment before delivery. The acquired experience confirm the utility of PPO in the ordinary clinical practice.