Influence of climatic parameters on the probabilistic design of green roofs.

Green roofs are effective tools for stormwater control in highly urbanized areas since they allow the reduction of peak runoffs and volumes discharged in sewer systems. Their design is quite standardized, except for the thickness of the growing medium layer, which is strictly related to vegetation type and rainfall regime. The paper proposes an analytical probabilistic approach that relates the climatic variables, the growing medium thickness, and the water content in the condition of fulfilled field capacity to the probability that runoff from green roofs exceeds a fixed threshold. The developed equations also consider the possibility of a reduced retention capacity due to previous rainfall events, that strongly influence the performance of these green infrastructures, especially when short dry periods and/or low evapotranspiration rates occur. This feature, neglected by the traditional design storm approach, and only partially considered by previous analytical probabilistic models, represent a great potentiality of the proposed equations that are also more user-friendly and less time-consuming than continuous simulation analysis. The focus of the paper is on the influence of climatic parameters on runoff probability. To this aim to perform the monthly analysis is fundamental, especially when there is a strong variability of the climatic parameters throughout the year. The model was tested in a case study in Milano, Italy. The application presented a good agreement between the results obtained from the proposed equations and those obtained from the continuous simulation of recorded data. The results also highlighted the importance of performing analysis on a monthly scale.

Efficacy and activity of PD-1 blockade in patients with advanced esophageal squamous cell carcinoma: a systematic review and meta-analysis with focus on the value of PD-L1 combined positive score.

BACKGROUND: Recently, several randomized controlled trials (RCTs) investigated immunotherapy-based regimens versus chemotherapy alone in patients with advanced esophageal squamous cell carcinoma (ESCC). Here we conducted a systematic review and meta-analysis on the efficacy and activity of programmed cell death protein 1 blockade in these patients, with focus on the value of programmed death-ligand 1 combined positive score (CPS) for selecting patients who may benefit the most. METHODS: RCTs investigating treatment with or without immune checkpoint inhibitors for advanced ESCC were selected. The hazard ratio (HR) and the odds ratio were used to compare the treatment effect on survival outcomes and tumor response, respectively, for immunotherapy-based regimens compared with standard chemotherapy, overall and according to geographic region or treatment line. We carried out a subgroup analysis comparing patients with CPS ≥10 or <10 and the evidence for treatment effect was evaluated by interaction test. RESULTS: A total of 5257 patients and 10 RCTs were included. Overall, the HR for overall survival benefit with immunotherapy-based regimens was 0.71 [95% confidence interval (CI) 0.66-0.76] compared with chemotherapy alone; such effect was independent from geographical region (Asia versus rest of the world) and treatment line (upfront versus second/further lines). The HR for progression-free survival benefit and the odds ratio for overall response rate increase were 0.78 (95% CI 0.66-0.93) and 1.50 (95% CI 1.22-1.83), respectively. The HR for overall survival benefit with immunotherapy-based treatment was 0.60 (95% CI 0.51-0.70) for CPS ≥10 subgroup versus 0.83 (95% CI 0.69-1.00) for CPS <10 (P for interaction 0.009). CONCLUSIONS: Immune checkpoint inhibitors have a consistent benefit in reducing the risk of death for ESCC patients which is dependent on programmed death-ligand 1 CPS status. Further investigations of biomarkers for immunotherapy in the subgroup of patients with CPS <10 are needed.

Clinical impact of neutropenia and febrile neutropenia in metastatic colorectal cancer patients treated with FOLFOXIRI/bevacizumab: a pooled analysis of TRIBE and TRIBE2 studies by GONO.

BACKGROUND: TRIBE and TRIBE-2 studies demonstrated higher benefit from FOLFOXIRI (fluorouracil, leucovorin, oxaliplatin, and irinotecan)/bevacizumab compared with FOLFIRI (fluorouracil, leucovorin, and irinotecan) or FOLFOX/bevacizumab as an upfront option for metastatic colorectal cancer patients, with more toxicities. We focused on the incidence and longitudinal dynamics of neutropenia and febrile neutropenia (FN) in the two studies, to evaluate their clinical relevance, the magnitude of impact of FOLFOXIRI/bevacizumab, and the role of risk factors in predicting their occurrence. METHODS: The overall incidence of grade 3-4 (G3-4) neutropenia and FN, the time to their onset, the use of granulocyte colony-stimulating factor, and the association with risk factors were evaluated in the overall population and according to treatment arm. FN episodes were assessed by Multinational Association for Supportive Care in Cancer (MASCC) score. RESULTS: Among 1155 patients, 568 (49%) received FOLFOXIRI/bevacizumab. Overall, 410 (35%) experienced G3-4 neutropenia and 70 (6%) FN, 21 (2%) at high risk. FOLFOXIRI/bevacizumab was associated with higher incidence of neutropenia (51% versus 21%, P < 0.001), FN (8% versus 4%, P = 0.02), and high-risk FN [18 (3%) versus 3 (1%), P = 0.015]. No related deaths were observed. The first episode of G3-4 neutropenia and FN occurred mainly in the first 2 months in both arms. Longitudinal analysis showed different patterns of evolution over cycles between the arms (P < 0.001) G3-4 neutropenia being more frequent in the first cycles with FOLFOXIRI/bevacizumab. Older patients (P = 0.01) and females (P < 0.001) had a significantly higher risk of G3-4 neutropenia. No significant interaction effect between arm and analysed risk factors in terms of risk of G3-4 neutropenia or FN was observed. The incidence of FN among older females receiving FOLFOXIRI/bevacizumab was 12%. Neither G3-4 neutropenia nor FN impaired efficacy in terms of overall response rate, progression-free survival, and overall survival. CONCLUSIONS: FOLFOXIRI/bevacizumab has a higher risk of G3-4 neutropenia and FN than doublets/bevacizumab. FN occurred in <10% of patients, mostly as low-risk episodes. A closer monitoring during the first 2 months is recommended; prophylactic use of granulocyte colony-stimulating factor may be considered for older females.

Impact of age and gender on the efficacy and safety of upfront therapy with panitumumab plus FOLFOX followed by panitumumab-based maintenance: a pre-specified subgroup analysis of the Valentino study.

BACKGROUND: The safety and efficacy outcome of elderly metastatic colorectal cancer (mCRC) patients fit enough to receive combination chemotherapy plus biological agents is an issue of growing interest. Also, gender-specific differential toxicity and efficacy of anti-epidermal growth factor receptor (EGFR)-based upfront treatments need to be explored. PATIENTS AND METHODS: Valentino was a multicenter, randomized, phase II trial, investigating two panitumumab-based maintenance strategies following first-line panitumumab plus FOLFOX in RAS wild-type mCRC patients. We carried out a subgroup analysis, aimed at assessing the differences in efficacy, safety and quality of life (QoL) according to age (<70 versus ≥70 years) and gender (male versus female). Efficacy endpoints were progression-free survival (PFS), overall survival (OS) and overall response rate (ORR); safety endpoints were rates of any grade and grade 3/4 adverse events (AEs). RESULTS: No significant differences in terms of PFS, OS and ORR were observed between patients aged <70 or ≥70 years and the effect of the maintenance treatment arm on survival outcomes was similar in the two subgroups. The safety profile of both induction and maintenance treatment and the impact on QoL were similar in elderly and younger patients. No significant differences in PFS, OS, ORR or clinical benefit rate were observed according to gender. A significantly higher rate of overall grade 3/4 AEs (P = 0.008) and of grade 3/4 thrombocytopenia (P = 0.017), any grade and grade 3/4 neutropenia (P < 0.0001) and any grade conjunctivitis (P = 0.033) was reported in female as compared to male patients. Conversely, we reported a significantly higher incidence of any grade skin rash (P = 0.0007) and hypomagnesemia (P = 0.029) in male patients. CONCLUSIONS: The upfront choice of an anti-EGFR-based doublet chemotherapy followed by a maintenance strategy represents a valuable option in RAS wild-type mCRC irrespective of gender and age, though a careful evaluation of patients to maximize the risk/benefit ratio is warranted.

Temozolomide and irinotecan (TEMIRI regimen) as salvage treatment of irinotecan-sensitive advanced colorectal cancer patients bearing MGMT methylation.

Background: Non-randomized studies showed that temozolomide (TMZ) achieves an average 10% response rate in heavily pretreated metastatic colorectal cancer (mCRC) patients with promoter methylation of the DNA repair gene O6-methylguanine-DNA methyltransferase (MGMT). In this phase II trial, irinotecan and temozolomide (TEMIRI) combination regimen was assessed in irinotecan-sensitive, MGMT methylated/microsatellite stable (MSS) pretreated mCRC patients. Patients and methods: Key inclusion criteria were centrally confirmed MGMT methylation by methylation-specific PCR, MSS mCRC, progression after at least two prior chemotherapy regimens for advanced disease and irinotecan-free interval >3 months. TEMIRI (TMZ 150 mg/m2 on days 1-5 plus irinotecan 100 mg/m2 on days 1, 15 q28 days) was administered for six cycles, followed by maintenance with TMZ. The primary end point was overall response rate (ORR). Exploratory translational analyses included MGMT immunohistochemistry (IHC) and methyl-BEAMing (MB). Results: Between December 2014 and June 2017, 25 patients were enrolled. The primary end point was met, since six patients achieved a partial response [ORR 24%, 95% confidence interval (CI) 11% to 43%]. At a median follow-up of 15.6 months, median progression-free survival (mPFS) and overall survival (mOS) were 4.4 and 13.8 months, respectively. Only four (16%) patients had ≥ grade 3 (CTCAE 4.0) adverse events. All patients whose cancer was MGMT-positive IHC were non-responders. Consistently, patients with MGMT-negative/low tumors had a significantly longer mPFS than others (6.9 versus 2.0 months; hazard ratio = 0.29, 95% CI 0.02-0.41; P = 0.003) and a non-significant trend for longer mOS. MB testing showed similar accuracy. Conclusions: TEMIRI regimen is a safe and active option in pre-treated, irinotecan-sensitive mCRC patients with MGMT methylation.

mTOR as a target of everolimus in refractory/relapsed Hodgkin lymphoma.

Despite impressive treatment advances, few options for refractory or relapsed Hodgkin Lymphoma (HL) are available and there is a need for new compounds development. A number of promising agents with multiple mechanisms of action are under investigation. Microenvironment and neoangiogenesis are acquiring a rising relevance in the pathophysiology and progression of HL. Everolimus (RAD001) is an oral antineoplastic agent derived from rapamycin, a macrocyclic lactone antibiotic, targeting the mammalian target of rapamycin (mTOR). Although the importance of mTOR signaling in the deregulated cell growth of human neoplastic cells has been recognized, this pathway is also emerging as a key regulator of the tumor response to hypoxia, as well as endothelial and stromal cells function, thereby regulating neoangiogenesis. Furthermore, mTOR plays an important role in anticancer drug resistance. The actions of everolimus within the mTOR pathway in HL result in decreased protein synthesis and cell cycle arrest, as well as in decreased angiogenesis. Everolimus has shown preliminary evidence of efficacy as a single-agent in heavily pretreated relapsed/refractory HL, with an overall fair safety profile. The purpose of this review is to discuss the employment of everolimus as an antiproliferative and antiangiogenic agent in HL and to report the critical role of the mTOR pathway and angiogenesis in this malignancy.

In vitro and in vivo activities of novel, semisynthetic thiopeptide inhibitors of bacterial elongation factor Tu.

Recently, we identified aminothiazole derivatives of GE2270 A. These novel semisynthetic congeners, like GE2270 A, target the essential bacterial protein elongation factor Tu (EF-Tu). Medicinal chemistry optimization of lead molecules led to the identification of preclinical development candidates 1 and 2. These cycloalklycarboxylic acid derivatives show activity against difficult to treat Gram-positive pathogens and demonstrate increased aqueous solubility compared to GE2270 A. We describe here the in vitro and in vivo activities of compounds 1 and 2 compared to marketed antibiotics. Compounds 1 and 2 were potent against clinical isolates of methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci (MIC(90) ≤ 0.25 µg/ml) but weaker against the streptococci (MIC(90) ≥ 4 µg/ml). Like GE2270 A, the derivatives inhibited bacterial protein synthesis and selected for spontaneous loss of susceptibility via mutations in the tuf gene, encoding EF-Tu. The mutants were not cross-resistant to other antibiotic classes. In a mouse systemic infection model, compounds 1 and 2 protected mice from lethal S. aureus infections with 50% effective doses (ED(50)) of 5.2 and 4.3 mg/kg, respectively. Similarly, compounds 1 and 2 protected mice from lethal systemic E. faecalis infections with ED(50) of 0.56 and 0.23 mg/kg, respectively. In summary, compounds 1 and 2 are active in vitro and in vivo activity against difficult-to-treat Gram-positive bacterial infections and represent a promising new class of antibacterials for use in human therapy.

Duropleural fistula revealed by neurological manifestations: an unusual cause of pleural effusion.

Symptomatic pleural collection of cerebrospinal fluid is a rare but accepted complication in hydrocephalic paediatric patients treated with ventriculopleural shunts. Few cases have been described in adults, usually as complication of trauma, tumours or spinal surgery, particularly post-laminectomy. It should be considered in the differential diagnosis of pleural effusion after neurosurgical procedures involving the spine. We describe two patients with large cerebrospinal fluid collections in the pleural cavity caused by postoperative duropleural fistula, who presented with neurological symptoms, cerebrospinal fluid pressure headache and meningitis.

Early aseptic loosening of a porous tantalum knee prosthesis.

Porous tantalum represents a relatively new solution for primary and revision total knee arthroplasty, offering several unmatched properties. Tantalum is a transition metal, with excellent biocompatibility and bioactivity due to its intrinsic physical and structural characteristics. A widespread clinical use of porous tantalum tibial components for primary total knee arthroplasty has been partly hindered by the difficulty in removing this type of implant after bone in growth, often leading to a significant bone defect. On the contrary, in the case here reported, removal of the trabecular metal tibial component was unexpectedly easy at a 7-month follow-up due to the absence of bone ingrowth but with a complete preservation of the tibial plate bone stock. Causes for the lack of bone ingrowth are discussed.

Topical kanamycin: an effective therapeutic option in aerobic vaginitis.

Eighty-one patients with clinical diagnosis of aerobic vaginitis (AV) were included in the study. The patients were randomized for treatment, 45 with kanamycin (100 mg vaginal ovules for 6 days, consecutively) and 36 with meclocycline (35 mg vaginal ovules for 6 days, consecutively). The patients were examined before starting the study, 1-2 days after treatment and 30 days after the end of the study. At the first follow-up the patients showed different levels of symptom reduction. Reduction in the presence of leukocytes, vaginal mucosa burning and itching were statistically significant in the group treated with kanamycin with respect to the group treated with meclocycline. Moreover, there was also reduced isolation of Enterobacteriaeae (97%) in the group treated with kanamycin versus those treated with meclocycline (76%). At the second follow-up, vaginal homeostasis (normalization of pH and presence of lactobacilli) was more evident in the kanamycin-treated group. In conclusion, our data suggest that the topical use of kanamycin could be considered a specific antibiotic for the therapy of this new pathology.

Fibroblast growth factor-2 expression during experimental oral carcinogenesis. Its possible role in the induction of pre-malignant fibrosis.

BACKGROUND: The fibroblastic growth factor (FGF)-2 has been shown to induce angiogenesis in several tumor types. To date, the activity of FGF during the development of oral pre-cancerous lesions has not been analyzed. We herein evaluated the role of FGF-2 in the pre-cancerous and cancerous lesions in the hamster cheek pouch oral cancer model. METHODS: Expression of FGF-2 and its receptors FGFR-2 and FGFR-3 was assessed by immunohistochemistry at different stages of the carcinogenesis protocol. Activity of FGF-2 isoforms was analyzed by Western blots. RESULTS: Increase and abnormal localization of FGF-2 expression was evident in cancerized epithelium before it was possible to detect morphologic alterations. The changes in FGF-2 are concomitant with the evolution of subepithelial fibrosis. Immunolabeling of carcinomas was faint or completely negative. Increases of FGF-2 activity are mainly due to the increase in the 18 kDa isoform. Receptors 2 and 3 of FGF are present in epithelium, fibroblasts, and vascular endothelia of control samples and in all stages of malignant transformation. CONCLUSIONS: Our results would suggest a role for FGF-2 in the epithelium-connective interactions and a deregulation of its expression in the early stages of oral cancerization. In pre-cancerous tissue FGF-2 would play a central role in the development of fibrosis and a more collateral role in the induction of angiogenesis. The data would indicate its involvement in the process via the 18 kDa isoform.

Ploidy analysis of field cancerization and cancer development in the hamster cheek pouch carcinogenesis model.

BACKGROUND: The hamster cheek-pouch carcinogenesis model is a well-known animal system that closely mimics the development of premalignant and malignant lesions in human oral cancer. Our aim was to numerically characterize the premalignant and malignant lesions and expressions of field cancerization in this model using ploidy as the end-point. METHODS: To study the DNA content and proliferation status of the cells in this model we assessed the Feulgen reaction and the immunohistochemical reaction for 5-bromo-2-deoxiuridine (BrdU) in different histological areas of serial tissue sections of the cheek pouches of animals injected with BrdU. RESULTS: Ploidy values were higher in cancerized epithelia with no unusual microscopic features (NUMF), in preneoplastic and tumor areas than in control epithelia. The aneuploidy index was higher in NUMF areas than in control and differed significantly from control in preneoplastic areas and carcinoma. CONCLUSIONS: The unexpected alteration in DNA content observed in NUMF epithelia is of great relevance as a biomarker of field cancerized areas.

Consenso argentino de ventilación no invasiva / Argentine consensus of non-invasive ventilation

Non-invasive ventilation (NIV) is nowadays increasingly used. The significant decrease in tracheal intubation related complications makes it particularly attractive in patients with moderately acute respiratory failure (ARF) who still have some degree of respiratory autonomy. It has also been used to support patients with chronic respiratory failure. However, final outcomes are variable according to the conditions which determined its application. This Consensus was performed in order to review the evidence supporting the use of positive pressure NIV. The patho-physiological background of NIV and the equipment required technology are described. Available evidence clearly suggests benefits of NIV in acute exacerbation of chronic obstructive pulmonary disease (COPD) and in cardiogenic pulmonary edema (Recommendation A). When considering ARF in the setting of acute respiratory distress syndrome results are uncertain, unless dealing with immunosupressed patients (Recommendation B). Positive results are also shown in weaning of mechanical ventilation (MV), particularly regarding acute exacerbation of COPD patients (Recommendation A). An improved quality of life in chronic respiratory failure and a longer survival in restrictive disorders has also been shown (Recommendation B) while its benefit in stable COPD patients is still controversial (Recommendation C). NIV should be performed according to pre-established standards. A revision of NIV related complications is performed and the cost-benefit comparison with invasive MV is also considered.

La ventilación no invasiva (VNI) ha alcanzado notable difusión en los últimos años. El ahorro delas complicaciones causadas por la intubación traqueal la hace especialmente atractiva en pacientesque presentan insuficiencia respiratoria aguda (IRA) no muy grave y que conservan cierta autonomía respiratoria.También se han descripto efectos terapéuticos en pacientes con insuficiencia respiratoria crónica de etiologías diversas. No obstante, los resultados obtenidos son variables según las circunstancias que motivan su aplicación. A fin de revisar la evidencia a favor de su uso se elaboró este Consenso referido particularmente a la VNI a presión positiva. Se describen su fundamento fisiopatológico, esencial para su correcta aplicación, y elequipamiento necesario para implementarla. La evidencia existente en la literatura establece definida utilidad de la VNI en la exacerbación de la EPOC y en el edema agudo de pulmón cardiogénico (Recomendación A).Su beneficio es aún incierto en pacientes con IRA secundaria a síndrome de dificultad respiratoria aguda, salvoen el subgrupo de pacientes inmunosuprimidos (Recomendación B). Los resultados son también favorablesen la desvinculación de la asistencia respiratoria mecánica (ARM), especialmente en pacientes ventilados por exacerbación de EPOC (Recomendación A). En la insuficiencia respiratoria crónica se ha hallado mejoría en la calidad de vida y mayor sobrevida en pacientes con enfermedades restrictivas (Recomendación B), mientrasque existe aún controversia sobre su utilidad en pacientes con EPOC estable (Recomendación C). La VNI debeser aplicada con estándares de cuidados que son establecidos. Se revisan las eventuales complicaciones derivadasde su uso y el beneficio costo-efectividad ahorrando recursos de mayor complejidad y disminuyendolos riesgos que implica la ARM invasiva.

Consenso argentino de ventilación no invasiva / Argentine consensus of non-invasive ventilation

Non-invasive ventilation (NIV) is nowadays increasingly used. The significant decrease in tracheal intubation related complications makes it particularly attractive in patients with moderately acute respiratory failure (ARF) who still have some degree of respiratory autonomy. It has also been used to support patients with chronic respiratory failure. However, final outcomes are variable according to the conditions which determined its application. This Consensus was performed in order to review the evidence supporting the use of positive pressure NIV. The patho-physiological background of NIV and the equipment required technology are described. Available evidence clearly suggests benefits of NIV in acute exacerbation of chronic obstructive pulmonary disease (COPD) and in cardiogenic pulmonary edema (Recommendation A). When considering ARF in the setting of acute respiratory distress syndrome results are uncertain, unless dealing with immunosupressed patients (Recommendation B). Positive results are also shown in weaning of mechanical ventilation (MV), particularly regarding acute exacerbation of COPD patients (Recommendation A). An improved quality of life in chronic respiratory failure and a longer survival in restrictive disorders has also been shown (Recommendation B) while its benefit in stable COPD patients is still controversial (Recommendation C). NIV should be performed according to pre-established standards. A revision of NIV related complications is performed and the cost-benefit comparison with invasive MV is also considered.(AU)

La ventilación no invasiva (VNI) ha alcanzado notable difusión en los últimos años. El ahorro delas complicaciones causadas por la intubación traqueal la hace especialmente atractiva en pacientesque presentan insuficiencia respiratoria aguda (IRA) no muy grave y que conservan cierta autonomía respiratoria.También se han descripto efectos terapéuticos en pacientes con insuficiencia respiratoria crónica de etiologías diversas. No obstante, los resultados obtenidos son variables según las circunstancias que motivan su aplicación. A fin de revisar la evidencia a favor de su uso se elaboró este Consenso referido particularmente a la VNI a presión positiva. Se describen su fundamento fisiopatológico, esencial para su correcta aplicación, y elequipamiento necesario para implementarla. La evidencia existente en la literatura establece definida utilidad de la VNI en la exacerbación de la EPOC y en el edema agudo de pulmón cardiogénico (Recomendación A).Su beneficio es aún incierto en pacientes con IRA secundaria a síndrome de dificultad respiratoria aguda, salvoen el subgrupo de pacientes inmunosuprimidos (Recomendación B). Los resultados son también favorablesen la desvinculación de la asistencia respiratoria mecánica (ARM), especialmente en pacientes ventilados por exacerbación de EPOC (Recomendación A). En la insuficiencia respiratoria crónica se ha hallado mejoría en la calidad de vida y mayor sobrevida en pacientes con enfermedades restrictivas (Recomendación B), mientrasque existe aún controversia sobre su utilidad en pacientes con EPOC estable (Recomendación C). La VNI debeser aplicada con estándares de cuidados que son establecidos. Se revisan las eventuales complicaciones derivadasde su uso y el beneficio costo-efectividad ahorrando recursos de mayor complejidad y disminuyendolos riesgos que implica la ARM invasiva.(AU)

In vitro sensitivity of human gastric cancer cells (HGC-27) to Helicobacter pylori cytotoxin.

BACKGROUND: The VacA cytotoxin produced by Helicobacter pylori is considered an important co-factor in the pathogenesis of chronic gastritis, peptic ulcer and gastric carcinoma. The toxin remains partly bound on the bacterial surface, but a certain amount is secreted and can bind receptors on gastric epithelium. The vacuolizing activity of this toxin is related to alteration of endo-lysosomic function and pore formation into plasmatic membrane. METHODS: We investigated the 'in vitro' effect of filtrates obtained from two broth cultures of H. pylori with different genotype (vacA+ and vacA-) as verified by PCR. The effect was studied on three cell lines of epithelial origin: HeLa cells (reference strain for testing vacuolization), human transformed keratinocytes HaCaT, human gastric carcinoma cells HGC-27, and on a murine leukaemia WEHI-3B. The filtrate concentrations capable of giving vacuolization (NRU test), antiproliferative and cytotoxic effects (MTT test) were determined. The modulating effect of filtrates on drug toxicity was investigated on HeLa and HGC-27 cells by testing topoisomerase inhibitors (Ciprofloxacin and Camptothecin) and non-steroidal anti-inflammatory molecules (Aspirin and Indomethacin). RESULTS: Our results confirm that vacuolizing activity is present only in VacA+ filtrate and that HaCaT and HeLa cells show a similar sensitivity, whereas gastric HGC-27 cells appear significantly resistant to VacA+ activity. Although VacA filtrate does not produce vacuolisation, it affects the cell proliferation and is cytotoxic to the four cell lines. Both the VacA+ and VacA- filtrates (at non-cytotoxic concentrations) produce a decrease in drug toxicity with the unique exception of Ciprofloxacin to gastric HGC-27 cells, which in the presence of VacA+ and VacA- produces a significant increase in toxicity. CONCLUSIONS: These data suggest that products from H. pylori (other than those that have antiproliferative and toxic activity) may modulate the sensitivity of cells to drugs 'in vitro'. If this also occurs 'in vivo', we can assume that H. pylori products interfere with drug activity on gastric tissue and also with other factors (such as cytokines) with a role in the genesis of diseases in which Helicobacters are potentially involved.

A new technique for staining mast cells using ferroin.

We describe here a new method for specific staining of mast cells using ferroin. Different hamster tissues were fixed in 4% formalin and processed for paraffin embedding. Sections were stained with hematoxylin followed by ferroin acidified with 2.5 N sulfuric acid to pH 4.0. Mast cells stained an intense orange color that contrasted markedly with bluish violet nuclei. High contrast was also observed when ferroin colored sections were counterstained with light green instead of hematoxylin. To evaluate the specificity of the stain, hamster cheek pouch sections were stained with toluidine blue, alcian blue-safranin O, and ferroin. Quantitative evaluation of mast cells stained with the three techniques showed no statistical difference. The simplicity and selectivity of this method is sufficient for image analysis of mast cells.

Selection criteria and long-term results of surgery in symptomatic rectocele.

BACKGROUND: The aim of this study was to confirm some selection criteria for the transrectal repair of the anterior rectocele and to compare our surgical results with those reported in the literature. METHODS: From January 1992 to December 1999, 30 females (mean age 52.9 years, range 28-70 yrs) affected by anterior rectocele were prospectively evaluated with a standard questionnaire, clinical examination, proctosigmoidoscopy, colonic transit time, dynamic defecography, anal EMG, anal manometry. Then, they were submitted to transrectal repair of rectocele with anterior plication of the rectal muscular wall. Fourteen (46.6%) of them were also submitted to perineal levatorplasty. Patients were followed postoperatively (mean 25.7 months) with the same standard questionnaire, clinical examination, defecography, and manometry. Results were tested by Fisher's Exact text, Wilcoxon's test, and "t"-test. RESULTS: Rectal dyschezia, incomplete evacuation, digital help in defecating, mean stool frequency, and rectal bleeding significantly improved. After 3 months, 30% of patients had no complaints, 40% had only 1-2 episodes/month complaints, 13.3% had evacuation only using laxatives, and 16.6% were unchanged. Defecography showed a significant reduction of the rectocele in 70% of patients after 3 months. Manometric parameters were not significantly modified. Four (28.6%) out of 14 patients submitted to perineal levatorplasty complained of dyspareunia. CONCLUSIONS: Our surgical results were comparable with those reported in the literature, with more than 80% of successful outcome. Preoperative clinical data and defecography were confirmed to be basic parameters in selecting patients for surgery. Colonic transit time, anal EMG, and anorectal manometry demonstrated to be useful to recognize conditions as slow colonic transit time, peripheral denervation, and reduced voluntary contraction that could lead to a less satisfactory outcome after surgery, and might benefit with a postoperative perineal rehabilitation by biofeedback and anal electrostimulations. The perineal levatorplasty is not suitable in young females, due to the risk of dyspareunia.