Antibiofilm effect of cleaning agents for ocular prostheses.

Introduction: This study aimed to evaluate the antibiofilm effect of different agents (neutral soap, 4% chlorhexidine, Efferdent effervescent tablets, 1% triclosan, and citronella essential oil) used for ocular prosthesis cleaning. Material and Methods: Biofilms of S. aureus and S. epidermidis were formed on 60 ocular prosthesis acrylic resin specimens. The specimens were cleaned with the studied agents with different techniques. Microorganism counting was performed. Data were submitted to ANOVA and HSD Tukey-Kramer (p<.01). Results: When compared to the control group, all cleaning protocols promoted a reduction in growth of microorganisms. The 4% chlorhexidine, effervescent tablets, and 1% triclosan cleaning agents eliminated biofilm in all groups. Conclusion: Therefore, immersion in 4% chlorhexidine, effervescent tablets, and 1% triclosan could be the best protocols indicated for ocular prosthesis cleaning due to their ability to eliminate biofilm.

Two-month stop in mammographic screening significantly impacts on breast cancer stage at diagnosis and upfront treatment in the COVID era.

INTRODUCTION: The present analysis aims to evaluate the consequences of a 2-month interruption of mammographic screening on breast cancer (BC) stage at diagnosis and upfront treatments in a region of Northern Italy highly affected by the severe acute respiratory syndrome-related coronavirus-2 (SARS-CoV-2) virus. METHODS: This retrospective single-institution analysis compared the clinical pathological characteristics of BC diagnosed between May 2020 and July 2020, after a 2-month screening interruption, with BC diagnosed in the same trimester of 2019 when mammographic screening was regularly carried out. RESULTS: The 2-month stop in mammographic screening produced a significant decrease in in situ BC diagnosis (-10.4%) and an increase in node-positive (+11.2%) and stage III BC (+10.3%). A major impact was on the subgroup of patients with BC at high proliferation rates. Among these, the rate of node-positive BC increased by 18.5% and stage III by 11.4%. In the subgroup of patients with low proliferation rates, a 9.3% increase in stage III tumors was observed, although node-positive tumors remained stable. Despite screening interruption, procedures to establish a definitive diagnosis and treatment start were subsequently carried out without delay. CONCLUSION: Our data showed an increase in node-positive and stage III BC after a 2-month stop in BC screening. These findings support recommendations for a quick restoration of BC screening at full capacity, with adequate prioritization strategies to mitigate harm and meet infection prevention requirements.

Onsite advanced biocleaning system for historical wall paintings using new agar-gauze bacteria gel.

AIMS: This study reports the results of the application of a new agar-gauze biogel system activated with viable bacterial cells to altered wall paintings. METHODS AND RESULTS: Biocleaning using agar biogel and agar-gauze biogel systems was performed onsite by direct application to altered wall painting surfaces (25-1000 cm2 ). The treatments were performed for the restoration of two original Italian sites: (i) at the Vatican Museums, Cristo che salva Pietro dalle acque-La Navicella, a wall painting by Giovanni Lanfranco (1627-1628) and (ii) at Pisa Cathedral Cupola, Incarnato, a wall painting by Orazio Riminaldi (1593-1630) and his brother Girolamo Riminaldi. The novelty of this study is the use of viable Pseudomonas stutzeri A29 cells in an advanced agar-gauze biogel system and the short bio-application contact times of between 3 and 12 h. The historical artworks were altered by lipid and protein residues from past restoration, as confirmed by Py-gas chromatography-mass spectrometry and FT-IR data. The effectiveness of the biological treatment was assessed, and general considerations were discussed. CONCLUSIONS: The short bio-application contact time of advanced agar-gauze gel activated with viable P. stutzeri cells makes this biotechnology promising as an alternative method to the traditional onsite cleaning techniques currently in use for altered historical wall paintings. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, we report for the first time the biocleaning of altered materials located in vertical and vaulted areas using agar-gauze biogel with short application times. These findings are of great significance for future restoration activities and are crucial for determining the best preservation strategies in this field.

Effect of photopolymerized glaze application on bacterial adhesion on ocular acrylic resin surfaces submitted to accelerated ageing.

Conditions of the acrylic resin (AR) surface, such as roughness, can promote a favourable environment for the adhesion of micro-organisms, even on the surface of ocular prostheses. This study evaluated the influence of photopolymerized glaze application on the roughness of ARs and adhesion of Staphylococcus aureus and Staphylococcus epidermidis on ocular AR surfaces submitted to accelerated ageing. Two hundred and eighty-eight samples of white colour (N1) and colourless ARs were distributed in eight groups (n = 9), based on surface treatments (glaze or ARs submitted to only a final polishing), accelerated ageing (before and after) and periods of microbial growth (24- and 48-h). The roughness average (Ra) and total height of roughness profile (Rt) values were greater for the groups with glaze and increased for all groups after ageing. The microbial adhesion among the groups with and without glaze did not present a statistically significant difference. The ageing did not statistically affect the adhesion of Staph. epidermidis, but affected the adhesion of Staph. aureus, which presented an increase after 24 h of growth on only N1 AR with glaze. These results demonstrate that the glaze did not contribute to adhesion of Staph. aureus and Staph. epidermidis, which are responsible for most ocular prosthetic infections. SIGNIFICANCE AND IMPACT OF THE STUDY: Some recent evidence suggested that the surface finish of ocular prostheses influences the accumulation of deposits that can affect the interaction with pathogenic bacteria, increasing the probability of infections. In addition, surface deterioration over time can increase the roughness and, consequently, biofilm formation. Thus, a better understanding of the influence of surface finish on bacterial adhesion becomes extremely important. In this study, we tested a glaze for surface polishing compared to mechanical polishing, before and after ageing. The results suggest that the glaze did not contribute to microbial adhesion and might be useful in preventing possible prosthetic infections.

Hi-tech restoration by two-steps biocleaning process of Triumph of Death fresco at the Camposanto Monumental Cemetery (Pisa, Italy).

AIMS: In this work, the 'hi-tech' complex biocleaning and restoration of the 14th-century fresco Triumph of Death (5·6 × 15·0 m) at the Camposanto Monumental Cemetery (Pisa, Italy) is reported. Since 2000, the restoration based on the biological cleaning of noble medieval frescoes, has been successfully utilized in this site. METHODS AND RESULTS: The novelty of this study is the two-steps biocleaning process using Pseudomonas stutzeri A29 viable cells, previously applied for recovering other valuable frescoes. In this case, after the fresco detachment from the asbestos-cement support (eternity), both the animal glue and the residues of calcium caseinate were biologically removed respectively from the front and from the back of the fresco in 3 h as indicated by GC-MS and PY/GC-MS analyses. The data obtained during the monitoring of the biorestoration process confirmed that the adopted procedure does not leave residual cells on the fresco surfaces as showed by plate count method, ATP determination and also SEM observation. In addition, to avoid the risk of condensation phenomena after the relocation of the restored fresco sections onto the original walls, the use of a new support has been set up together with the design of a control system that allows a continuous monitoring of environmental parameters for prevention and conservation purposes. CONCLUSIONS: This large-scale biorestoration work clearly shows and confirms that this biotechnology is highly efficient, safe, noninvasive, risk-free and very competitive compared to the traditional cleaning methods, offering an unusual 'resurrection' of the degraded artworks also in very complicated and delicate conditions such as the Triumph of Death fresco, defined for its dimension and artistic importance the 'Pisa's Sistina frescoes'. SIGNIFICANCE AND IMPACT OF THE STUDY: These findings can be of significant importance for other future new restoration activities and they are crucial for determining preservation strategies in this field.

Impact of time to surgery after neoadjuvant chemotherapy in operable breast cancer patients.

BACKGROUND: The optimal time interval between the end of neoadjuvant systemic therapy (NST) and breast surgery is still unclear. It is not known if a delay in surgery might influence the benefit of primary chemotherapy. The aim of this study is to evaluate the relationship between time to surgery (TTS) and survival outcomes. PATIENTS AND METHODS: According to TTS, women with diagnosis of BC treated with NST were divided into two cohorts: group A = 21 days or fewer and group B = longer than 21 days. OS and RFS were estimated and compared according to TTS and known prognostic factors. RESULTS: A total of 319 patients were included in the study: 61 in group A and 258 in group B. Median TTS was 34 days. No association between clinical stage, nuclear grade, type of chemotherapy, type of surgery and TTS was detected. OS and RFS were significantly worse for group B compared with group A, with a hazard ratio of 3.1 (95% CI, 1.1-8.6 p = 0.03) and 3.1 (95% CI, 1.3-7.1 p = 0.008) respectively. Multivariate analysis confirmed that TTS was an independent prognostic factor in term of OS (p = 0.03) and RFS (p = 0.01). Even in the subgroup of patients with pCR, TTS continued to be an independent prognostic factor for both OS and RFS (p = 0.05 and p = 0.03). CONCLUSIONS: TTS after NST seems to influence survival outcomes. BC patients underwent surgery within 21 days experienced maximal benefit from previous treatment: this advantage is consistent and maintained over time.

Rare cases reports of gastrointestinal stromal tumour (GIST).

The GISTs are rare tumours but even rarer is the localization in some districts. We reported two GISTs of the duodenum, two of the omentum and peritoneum, one of the rectum and one of a Meckel's diverticulum. These exceptional locations are confirmed by the relative difficult diagnosis, obtained in some cases only by the surgical treatment despite the CT and MR. The endoscopy is useful in hemorrhagic and duodenum forms, only for the diagnosis and for the control of blood loss. Surgical treatment in all cases was decisive without the need to make use of adjuvant therapy, with positive long-term results, which excluded the disappearance of relapses or secondary lesions.

A rapid genetic counselling and testing in newly diagnosed breast cancer is associated with high rate of risk-reducing mastectomy in BRCA1/2-positive Italian women.

BACKGROUND: Risk-reducing mastectomy (RRM) decreases breast cancer (BC) risk in BRCA1/2 mutation carriers by up to 95%, but the Italian attitude towards this procedure is reluctant. PATIENTS AND METHODS: This is an observational study with retrospective design, using quantitative and qualitative research methods, aimed at evaluating the attitude towards RRM by rapid genetic counselling and testing (RGCT), at the time of BC diagnosis, compared with traditional genetic counselling and testing (TGCT), after previous BC surgery. Secondary aims were to investigate patient satisfaction after RRM and the rate of occult tumour in healthy breasts. A total of 1168 patients were evaluated: 1058 received TGCT, whereas 110 underwent RGCT. RESULTS: In TGCT, among 1058 patients, 209 (19.7%) mutation carriers were identified, with the rate of RRM being 4.7% (10 of 209). Conversely in RGCT, among 110 patients, 36 resulted positive, of which, 15 (41.7%) underwent bilateral mastectomy at the BC surgery time, showing an overall good satisfaction, measured by interpretative phenomenological analysis 12 months after the intervention. CONCLUSIONS: Our study shows that RGCT in patients with a hereditary profile is associated with a high rate of RRM at the BC surgery time, this being the pathway offered within a multidisciplinary organization.

Multi-technique study of a ceramic archaeological artifact and its content.

In this paper we report the results of a study performed with different physical and chemical methods on a ceramic vase originally attributed to the I century CE. The joint use of infrared spectroscopic analysis and laser techniques, as well as pyrolysis-gas chromatography coupled with mass spectrometry and thermoluminescence, allowed us to characterize the vase material and its content. The chemical data were combined with morphological and stylistic examinations of the object and helped in defining its actual geographical and chronological pertinence.

Laparoscopic treatment of paraesophageal and mixed diaphragmatic hernias. Report of two cases.

Hiatal hernias are classified into 3 types: sliding hernia (type I), paraesophageal hernia (type II) and mixed hernia (type III), that is a combination of type I and II. The paraesophageal and mixed hernias represent about 5-10% of the surgically treated hiatal hernias. The surgical treatment of the paraesophageal and mixed hernias is unavoidable because of the high risk of severe complications and it has to be considered in a high percentage of cases. The most important technical difficulty in the video-laparoscopic treatment is represented by the hugeness of the hernial defect and by the challenging reduction of the stomach into the abdomen. A cautious dissection of hernial sac and diaphragmatic cruses as well as a careful crural repair make the video-laparoscopic procedure feasible. The operative times are not prolonged and the results are similar to the open technique ones. In literature, the incidence of both intra and postoperative complications doesn't exhibit statistically significant differences between laparoscopic and open techniques. Because of the complexity of the laparoscopic procedure, the minimally invasive access has to be reserved to surgeons who are well trained in those techniques. In this paper we describe 2 cases: one of paraesophageal hernia and the other of mixed hernia which were video-laparoscopically treated with the help, in the second case, of a Gore-Tex mesh. In both cases the technical results were positive. Intra and postoperative complications didn't occur and, one year after the surgical procedure, both patients were in good health and recurrence-free.

Analytical investigation of calcium oxalate films on marble monuments.

This paper describes an analytical approach to investigate the origin of oxalate films on marble. Calcium oxalate films were collected on buildings of historical importance in Lucca and Pisa (Italy) and characterised by optical microscopy (OM), scanning electron microscopy-energy dispersive X-ray spectrometry (SEM-EDX), Fourier transform infrared (FTIR) spectroscopy (equipped with diamond cell), and gas chromatography-mass spectrometry (GC-MS). The morphology of the films was investigated by optical and electronic microscopy. FTIR analyses highlighted the presence of calcium oxalate (both as whewellite and weddellite), gypsum, calcite, nitrates, silicates and apatite, while EDX maps showed the distribution of elements. Several samples showed traces of organic compounds, identified by GC-MS as paraffin wax, lipids of animal origin and egg. The correlation between organic material and oxalate contents suggests the origin of the films from degradation processes of past surface treatment.

Does solvent influence the ground-state tautomeric population of hypericin?

Nuclear magnetic resonance measurements indicate that hypericin exists in the same "normal" tautomeric form irrespective of whether the solvent is dimethyl sulfoxide or tetrahydrofuran. This result is discussed in the context of previous experimental and theoretical work. It is concluded that solvent perturbations cannot induce tautomerization in hypericin.

Conformational changes in pediocin AcH upon vesicle binding and approximation of the membrane-bound structure in detergent micelles.

Pediocin AcH is a 44-residue antimicrobial peptide with bactericidal potency against Gram-positive bacteria such as Listeria. It belongs to a family of bacteriocins that, when membrane-associated, is predicted to contain beta-sheet and alpha-helical regions. All bacteriocins in this family have a conserved N-terminal disulfide bond. An additional C-terminal disulfide bond in pediocin AcH is thought to confer enhanced potency and broader specificity range against sensitive bacteria. The C-terminal disulfide bond may also affect the conformation of the C-terminus. The secondary structures of pediocin AcH in aqueous solution and vesicles from susceptible cells, as well as the ability of trifluoroethanol (TFE) and detergent systems to induce secondary structures like those induced in vesicles, were studied by circular dichroism (CD) spectroscopy. Like related peptides, pediocin AcH was highly unordered in aqueous solution, 56%. However, it also contained 20% beta-strand and 15% beta-turn structures. Upon complete binding to vesicles, 32% alpha-helical structure formed, the unordered structure decreased to 32%, and the beta-strand and beta-turn structures remained largely unchanged. Thus, a betaalpha domain structure formed in vesicles. The helical structure likely forces the C-terminal tail to loop back on the helix so that the C24-C44 disulfide bond can form. Detergent micelles were superior to TFE in their ability to induce secondary structural fractions in pediocin AcH comparable to those observed in vesicles. This demonstrates the importance of a hydrocarbon-water interface to pediocin AcH structure induction and suggests that it is preferable to use detergent micelles as solvents in NMR studies of pediocin AcH structure.

A specific intermolecular association between the regulatory domains of a Tec family kinase.

Interleukin-2 tyrosine kinase (Itk), is a T-cell specific tyrosine kinase of the Tec family. We have examined a novel intermolecular interaction between the SH3 and SH2 domains of Itk. In addition to the interaction between the isolated domains, we have found that the dual SH3/SH2 domain-containing fragment of Itk self-associates in a specific manner in solution. Tec family members contain the SH3, SH2 and catalytic domains common to many kinase families but are distinguished by a unique amino-terminal sequence, which contains a proline-rich stretch. Previous work has identified an intramolecular regulatory association between the proline-rich region and the adjacent SH3 domain of Itk. The intermolecular interaction between the SH3 and SH2 domains of Itk that we describe provides a possible mechanism for displacement of this intramolecular regulatory sequence, a step that may be required for full Tec kinase activation. Additionally, localization of the interacting surfaces on both the SH3 and SH2 domains by chemical shift mapping has provided information about the molecular details of this recognition event. The interaction involves the conserved aromatic binding pocket of the SH3 domain and a newly defined binding surface on the SH2 domain. The interacting residues on the SH2 domain do not conform to the consensus motif for an SH3 proline-rich ligand. Interestingly, we note a striking correlation between the SH2 residues that mediate this interaction and those residues that, when mutated in the Tec family member Btk, cause the hereditary immune disorder, X-linked agamaglobulinemia.

[Gallbladder carcinoma. Personal case histories]. / Carcinoma della colecisti. Casistica personale.

BACKGROUND AND AIM: Carcinoma of the gallbladder is a rare pathology which is extremely malignant with a fatal prognosis owing to its difficult and late diagnosis. It represents 3% of all digestive tumours and is predominant in females. The major risk factor is gallbladder stones. METHODS: A total of 14 gallbladder neoplasms were diagnosed in 798 cholecystectomies performed from October 1990 to October 1995, with a male/female ratio of 6: 1 and a mean age of 68 years and 11 months. Biliary colic and jaundice represented approximately 93% of symptoms present in this series. Fourteen cholecystectomies were performed in association with other forms of surgery (1 GEA, 2 drainage acc. Kehr, 1 hemicolectomy, 1 resection of the hepatic bed, 1 hepaticojejunostomy). RESULTS: The operative mortality was nil. Mean survival was 1 year, 4 months and 15 days, and was obviously higher in the early stages. Four patients (all female) are still alive (mean survival 2 years 10 months 9 days). The five-year survival rate reported in the literature does not exceed 5-10%. Surgery does not currently give satisfactory results. In the light of present knowledge it is important to intervene on the risk factors in the form of laparoscopic cholecystectomies for patients with lithiasis or gallbladder polyps. CONCLUSIONS: Diagnosis must be as early as possible during the "early cancer" phase in order to ensure that surgery is oncologically radical and therefore also remedial.

Elevated insulin levels contribute to the reduced growth hormone (GH) response to GH-releasing hormone in obese subjects.

We have recently presented experimental evidence indicating that insulin has a physiologic inhibitory effect on growth hormone (GH) release in healthy humans. The aim of the present study was to determine whether in obesity, which is characterized by hyperinsulinemia and blunted GH release, insulin contributes to the GH defect. To this aim, we used a simplified experimental protocol previously used in healthy humans to isolate the effect of insulin by removing the interference of free fatty acids (FFAs), which are known to block GH release. Six obese subjects (four men and two women; age, 30.8 +/- 5.2 years; body mass index, 36.8 +/- 2.8 kg/m2 [mean +/- SE]) and six normal subjects (four men and two women; age, 25.8 +/- 1.9 years; body mass index, 22.7 +/- 1.1 kg/m2) received intravenous (i.v.) GH-releasing hormone (GHRH) 0.6 microg/kg under three experimental conditions: (1) i.v. 0.9% NaCl infusion and oral placebo, (2) i.v. 0.9% NaCl infusion and oral acipimox, an antilipolytic agent able to reduce FFA levels (250 mg at 6 and 2 hours before GHRH), and (3) euglycemic-hyperinsulinemic clamp (insulin infusion rate, 0.4 mU x kg(-1) x min(-1)). As expected, after placebo, the GH response to GHRH was lower for obese subjects versus normals (488 +/- 139 v 1,755 +/- 412 microg/L x 120 min, P < .05). Acipimox markedly reduced FFA levels and produced a mild reduction of insulin levels; under these conditions, the GH response to GHRH was increased in both groups, remaining lower in obese versus normal subjects (1,842 +/- 360 v 4,871 +/- 1,286 microg/L x 120 min, P < .05). In both groups, insulin infusion yielded insulin levels usually observed under postprandial conditions and reduced circulating FFA to the levels observed after acipimox administration. Again, the GH response to GHRH was lower for obese subjects versus normals (380 +/- 40 v 1,075 +/- 206 microg/L x 120 min, P < .05), and in both groups, it was significantly lower than the corresponding response after acipimox. In obese subjects, as previously reported in normals, the GH response to GHRH was inversely correlated with the mean serum insulin (r = -.70, P < .01). In conclusion, our data indicate that in the obese, as in normal subjects, the GH response to GHRH is a function of insulin levels. The finding that after both the acipimox treatment and the insulin clamp the obese still show higher insulin levels and a lower GH response to GHRH than normal subjects suggests that hyperinsulinemia is a major determinant of the reduced GH release associated with obesity.

Evidence for an inhibitory effect of physiological levels of insulin on the growth hormone (GH) response to GH-releasing hormone in healthy subjects.

It has been previously reported that in healthy subjects, the acute reduction of free fatty acids (FFA) levels by acipimox enhances the GH response to GHRH. In the present study, the GH response to GHRH was evaluated during acute blockade of lipolysis obtained either by acipimox or by insulin at different infusion rates. Six healthy subjects (four men and two women, 25.8 +/- 1.9 yrs old, mean +/- SE) underwent three GHRH tests (50 micrograms iv, at 1300 h) during: 1) iv 0.9% NaCl infusion (1200-1500 h) after oral acipimox administration (250 mg) at 0700 h and at 1100 h; 2) 0.1 mU.kg-1.min-1 euglycemic insulin clamp (1200-1500 h) after oral acipimox administration (250 mg at 0700 h and at 1100 h); 3) 0.4 mU.kg-1.min-1 euglycemic insulin clamp (1200-1500 h) after oral placebo administration (at 0700 and 1100 h). Serum insulin (immunoreactive insulin) levels were significantly different in the three tests (12 +/- 2, 100 +/- 10, 194 +/- 19 pmol/L, P < 0.06), plasma FFA were low and similar (0.04 +/- 0.003, 0.02 +/- 0.005, 0.02 +/- 0.003, not significant), and the GH response to GHRH was progressively lower (4871 +/- 1286, 2414 +/- 626, 1076 +/- 207 micrograms/L 120 min), although only test 3 was significantly different from test 1 (P < 0.05). Pooling the three tests together, a significant negative regression was observed between mean serum immunoreactive insulin levels and the GH response to GHRH (r = -0.629, P < 0.01). Our results indicate that in healthy subjects, acipimox and hyperinsulinemia produce a similar decrease in FFA levels and that at similar low FFA, the GH response to GHRH is lower during insulin infusion than after acipimox. These data suggest that insulin exerts a negative effect on GH release. Because the insulin levels able to reduce the GH response to GHRH are commonly observed during the day, for instance during the postprandial period, we conclude that the insulin negative effect on GH release may have physiological relevance.