Biological activity of roots and aerial parts of Zinnia peruviana on pathogenic micro-organisms in planktonic state and biofilm forming.

Microbial resistance to antibiotics affects the control of clinical infections and is a growing concern in global public health. One important mechanism whereby micro-organisms acquire resistance is biofilm formation. This context has led to the investigation of new antimicrobial substances from plants popularly used in folk medicine. In this work, we studied the antimicrobial and antibiofilm activity of Zinnia peruviana roots, ziniolide (major root metabolite) and aerial parts against Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, Escherichia coli, Pseudomonas aeruginosa and Candida albicans. The minimum inhibitory and minimum microbicidal concentration and inhibition of biofilm production was determined. All Z. peruviana extracts showed antimicrobial activity, but that corresponding to the roots was the most active one. The best inhibitory and microbicidal activity was detected against Gram-positive bacteria (0·039-0·078 mg ml-1 ). The acetonic extract from Z. peruviana leaves showed moderate activity against Gram-positive bacteria (0·625 mg ml-1 ). Acetonic extract of Z. peruviana flowers showed weak activity (1·25-5 mg ml-1 ). All the extracts tested showed inhibition of biofilm formation, as well as the ziniolide, however, roots and flowers extracts showed higher antibiofilm activity particularly against Staphylococcus, Listeria and Candida. The extracts tested may be a promising natural alternative for the control of microbial infections.

Transanal endoscopic microsurgery after neoadjuvant radiochemotherapy for locally advanced extraperitoneal rectal cancer.

PURPOSE: The aim of this study is to provide a prospective analysis of post-operative and oncological outcomes in patients affected by locally advanced rectal cancer (LARC), who obtained a major/complete clinical response after pre-operative radio-chemotherapy (RCT) and were treated with local excision (LE) by trans-anal endoscopic microsurgery (TEM) to confirm a pathological complete response (pCR) after to neo-adjuvant RCT. METHODS: All patients with LARC treated by pre-operative RCT and full-thickness LE by TEM (2000-2014) were included in the study. If the pathological analysis confirmed near complete or pCR, intensive follow up was proposed. If the pathological response was incomplete, a radical resection with TME was proposed. Post-operative (according to Clavien's classification), functional and long-term oncological outcome were analyzed. RESULTS: 36 patients were treated by TEM. The median post-operative hospital stay was 5 days. The post-operative morbidity was 41.6% (no grade ≥3). At pathological analysis, 23 specimens were ypT0 TRG1, and 4 were ypT1 TRG2. In 9 cases (ypT>1 and/or TRG>2), radical surgery with TME was proposed but 3 refused it. Median follow-up was 68 months. One local recurrence and 4 distant metastases occurred. The 5-yr actuarial local control, overall survival and disease-free survival were 96.0%, 92.0% and 82.8%. CONCLUSIONS: In case of major or complete clinical response of LARC after pre-operative RCT, LE by TEM can be used to confirm the pathological response. This avoids the necessity of radical surgery and, in our experience, this approach seems to guarantee oncological safety with the functional advantages of an organ-sparing procedure.

Evaluation of cytotoxicity and genotoxicity of Acacia aroma leaf extracts.

Acacia aroma, native plant from San Luis, Argentina, is commonly used as antiseptic and for healing of wounds. The present study was conducted to investigate the in vitro cytotoxicity and genotoxicity of hot aqueous extract (HAE) and ethanolic extract (EE) of A. aroma. The cytotoxic activity was assayed by neutral red uptake assay on Vero cell. Cell treatment with a range from 100 to 5000 µg/mL of HAE and EE showed that 500 µg/mL and 100 µg/mL were the maximum noncytotoxic concentrations, respectively. The CC50 was 658 µg/mL for EE and 1020 µg/mL for HAE. The genotoxicity was tested by the single-cell gel electrophoresis comet assay. The results obtained in the evaluation of DNA cellular damage exposed to varied concentrations of the HAE showed no significant genotoxic effect at range of 1-20 mg/mL. The EE at 20 mg/mL showed moderate genotoxic effect related to the increase of the DNA percentage contained in tail of the comet; DNA was classified in category 2. At concentrations below 5 mg/mL, the results of cytotoxicity and genotoxicity of aqueous and ethanolic extracts of Acacia aroma guarantee the safety at cell and genomic level. However further studies are needed for longer periods including animal models to confirm the findings.

Transanal endoscopic microsurgery after neoadjuvant radiochemotherapy for locally advanced extraperitoneal rectal cancer: short-term morbidity and functional outcome.

BACKGROUND: Transanal endoscopic microsurgery (TEM) after radiochemotherapy (RCT) has been reported in selected cases of locally advanced rectal cancer as an alternative to traditional radical resection with total mesorectal excision with a curative intent or as diagnostic tool to confirm a pathological complete response of the primary tumor. No study has evaluated functional outcome after TEM in preoperatively irradiated patients. METHODS: This study was designed to evaluate short-term morbidity (according to Clavien's classifications) and establish (by a questionnaire) continence and evacuative function after RCT and TEM, at 1 year from surgery, analyzing the impact of RCT on postoperative outcomes. Patients with locally advanced rectal cancer treated by RCT and TEM (group 1) or with early T1 or adenomas treated only by TEM (group 2) entered this cohort comparative study. RESULTS: Twenty-two patients entered the study as group 1 and 25 as group 2. No postoperative mortality occurred. The morbidity rate was 36.4 % in group 1 vs. 16 % in group 2 (p = 0.114). The rate of suture dehiscence was 22.7 % in group 1 vs. 4 % in group 2 (p = 0.068). No grade III complications, reoperation, or hospital readmission within 30 days was recorded in either group. One year after surgery, continence and evacuative scores in group 1 were 1.05 ± 1.25 and 24.72 ± 2.79, respectively, which were similar to group 2 (p = 0.081 and 0.288, respectively). CONCLUSIONS: TEM after RCT in selected rectal cancer patients has an acceptable morbidity and functional results at 1 year from surgery. Preoperative irradiation could increase postoperative short-term morbidity, but it does not seem to influence evacuative or sphincter function after 1 year from surgery.

Antibacterial activity of extracts of Acacia aroma against methicillin-resistant and methicillin-sensitive Staphylococcus

Antibacterial activity of organic and aqueous extracts of Acacia aroma was evaluated against methicillin-resistant Staphylococcus aureus (MRSA), methicillin sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus epidermidis. Inhibition of bacterial growth was determined using agar diffusion and bioautographic methods. Among all assayed organic extracts only ethanolic and ethyl acetate extracts presented highest activities against all tested Staphylococcus strains with minimal inhibitory concentration (MIC) values ranging from 2.5 to 10 mg/ml and from 2.5 to 5 mg/ml respectively. The aqueous extracts show little antibacterial activity against Staphylococcus strains. The bioautography assay demonstrated well-defined growth inhibition zones against S. aureus in correspondence with flavonoids and saponins. A. aroma would be an interesting topic for further study and possibly for an alternative treatment for skin infections.

Surgical treatment of small bowel neoplasms.

Small intestinal neoplasms are uncommon cancers. Benign small intestinal tumors (e.g., leiomyoma, lipoma, hamartoma, or desmoid tumor) usually are asymptomatic but may present with complications. Primary malignancies of the small intestine, including adenocarcinoma, leiomyosarcoma, carcinoid, and lymphoma, are often symptomatic and may present with intestinal obstruction, jaundice, bleeding, or pain. Metastatic neoplasms may involve the small intestine via contiguous spread, peritoneal metastases or hematogenous metastases. Because the small intestine is relatively inaccessible to routine endoscopy, diagnosis of small intestinal neoplasms is often delayed for months after onset of symptoms. During last years the increase of small bowel endoscopy and other diagnostic tools allow earlier non-operative diagnosis. Even though radical resection of small bowel cancer plays an important role, the 5 yr overall survival remains low.

Antibacterial activity of extracts of acacia aroma against methicillin-resistant and methicillin-sensitive Staphylococcus.

Antibacterial activity of organic and aqueous extracts of Acacia aroma was evaluated against methicillin-resistant Staphylococcus aureus (MRSA), methicillin sensitive Staphylococcus aureus (MSSA) and methicillin-resistant Staphylococcus epidermidis. Inhibition of bacterial growth was determined using agar diffusion and bioautographic methods. Among all assayed organic extracts only ethanolic and ethyl acetate extracts presented highest activities against all tested Staphylococcus strains with minimal inhibitory concentration (MIC) values ranging from 2.5 to 10 mg/ml and from 2.5 to 5 mg/ml respectively. The aqueous extracts show little antibacterial activity against Staphylococcus strains. The bioautography assay demonstrated well-defined growth inhibition zones against S. aureus in correspondence with flavonoids and saponins. A. aroma would be an interesting topic for further study and possibly for an alternative treatment for skin infections.

The role of local excision in rectal cancer after complete response to neoadjuvant treatment.

Correlation between pathological response of primary tumour and mesorectal lymph node involvement was prospectively evaluated to assess the role of local excision (LE) in rectal cancer after complete response to neoadjuvant treatment. A series of 272 consecutive rectal cancer, submitted to neoadjuvant radiochemotherapy (RCT) and surgery with total mesorectal excision (TME) were analysed. Tumour downstaging (pT) and tumour regression grade (TRG) together with sex, age, location of the tumour, pre-treatment clinical stage, type of chemoradiation and operation performed entered in an univariate and multivariate analysis. Pathological complete response on primary tumour was found in 56 patients (20.6%). Lymph node metastases were found in 72 patients (26.5%). The rate of positive nodes was 1.8% for pT0 and TRG1 cases, respectively, to go up to 6.3% for pT1 and 24.1% for TRG 2 cases, respectively. At the univariate analysis, factors with a statistically significant correlation with the risk of lymph node metastasis were: clinical pre-treatment N stage (p<0.05), pT stage (p<0.001) and TRG (p<0.001). At the multivariate analysis, the best predictors of pathologic lymph node involvement were pT stage (p=0.0013 ) and TRG (p=0.0011). Because LE is an adequate technique to assess the tumour pathological response and nodal involvement in pT0 or TRG1 cases seems extremely infrequent, radical resection is probably not justified after pathological complete response. Prospective randomized trials are necessary to establish if, in these cases, LE can guarantee the same oncologic results offered by the currently adopted protocols of RCT followed by radical resections.

Functional results after radiochemotherapy and total mesorectal excision for rectal cancer.

BACKGROUND AND PURPOSE: The aim of this study was to prospectively define and measure evacuation and continence disorders after preoperative radiochemotherapy and total mesorectal excision (TME) for rectal cancer 1 year after surgery. MATERIALS AND METHODS: We submitted 100 patients, who underwent neoadjuvant treatment and anterior resection with TME from 1996 to 2003, to a questionnaire on postoperative continence and evacuation. Anal sphincter function was further assessed by the Memorial Sloan-Kettering score. Factors influencing anorectal function were examined in univariate and multivariate analysis. RESULTS: Median evacuation score was 16.12 +/- 5.12 (range 0-28). Sensation of incomplete evacuation was reported in 58% of cases, necessity to return to the bathroom <15 min in 37% and inability to evacuate completely <15 min in 35%. Median continence score was 13.7 +/- 4.79 (range 0-20). Incontinence to flatus was reported in 46% of cases. Colonic J-pouch allows better evacuation and continence. Continence was also better in absence of postoperative complications. Sphincter function resulted excellent or good in 75% of patients according to the Memorial Sloan-Kettering score. CONCLUSIONS: The most frequent symptoms in our series are the sensation of incomplete evacuation, the incontinence to flatus, and the necessity to return to the bathroom <15 min. Colonic J-pouch warrants a better function. Postoperative complications compromise good functional results.

Genotypic, phenotypic, and clinical characteristics of isolates of Helicobacter pylori from San Luis, Argentina.

The vacA and cagA genotypes of Helicobacter pylori exhibited distinct geographic distribution and correlation with severity of disease. In the above genotypes (obtained from 150 H. pylori-positive patients--139 with gastritis, 10 with ulcer and 1 patient with gastric cancer) combinations vacA s1/m1 and s2/m2 were detected using PCR in 75 and 25% of isolates, respectively, in patients with chronic gastritis. The of s1/m1 and s2/m2 combinations were also detected from ulcers (60 and 40%, respectively). The cagA was detected in 30% of isolates. Concentrated culture supernatants of 7 (64%) out of 11 H. pylori strains induced vacuolization in Vero cells in titers ranging from 1:5 to 1:40. The vacA s1 genotype was significantly associated with, but not predictive of the presence of vacuolating cytotoxin activity and the cagA gene.

DNA fingerprinting by ERIC-PCR for comparing Listeria spp. strains isolated from different sources in San Luis, Argentina.

In this study, a total of 24 Listeria spp. strains were analyzed. Twenty-two isolates were obtained in San Luis (Argentina) from human, animal, and food samples. Two types of strains, Listeria monocytogenes CLIP 22762 and Listeria innocua CLIP 74915, were included as reference strains. All isolates were biochemically identified and characterized by serotyping, phage typing, and amplification of the flaA gene by polymerase chain reaction (PCR). Repetitive intergenic consensus (ERIC) sequence-based PCR was used to generate DNA fingerprints. On the basis of ERIC-PCR fingerprints, Listeria spp. strains were divided into three major clusters matching origin of isolation. ERIC-PCR fingerprints of human and animal isolates were different from those of food isolates. In addition, groups I and II included ten L. monocytogenes strains, and only one Listeria seeligeri strain. Group III included nine L. innocua strains and four L. monocytogenes strains. Computer evaluation of ERIC-PCR fingerprints allowed discrimination between the tested serotypes 1/2b, 4b, 6a, and 6b within each major cluster. The index of discrimination calculated was 0.94. This study suggests that the ERIC-PCR technique provides an alternative method for the identification of Listeria species and the discrimination of strains within one species.

DNA fingerprinting by ERIC-PCR for comparing Listeria spp. strains isolated from different sources in San Luis: Argentina / Caracterización molecular por ERIC-PCR de cepas de Listeria spp. aisladas de diversos orígenes en San Luis: Argentina

In this study, a total of 24 Listeria spp. strains were analyzed. Twenty-two isolates were obtained in San Luis (Argentina) from human, animal, and food samples. Two types of strains, Listeria monocytogenes CLIP 22762 and Listeria innocua CLIP 74915, were included as reference strains. All isolates were biochemically identified and characterized by serotyping, phage typing, and amplification of the flaA gene by polymerase chain reaction (PCR). Repetitive intergenic consensus (ERIC) sequence-based PCR was used to generate DNA fingerprints. On the basis of ERIC-PCR fingerprints, Listeria spp. strains were divided into three major clusters matching origin of isolation. ERIC-PCR fingerprints of human and animal isolates were different from those of food isolates. In addition, groups I and II included ten L. monocytogenes strains, and only one Listeria seeligeri strain. Group III included nine L. innocua strains and four L. monocytogenes strains. Computer evaluation of ERIC-PCR fingerprints allowed discrimination between the tested serotypes 1/2b, 4b, 6a, and 6b within each major cluster. The index of discrimination calculated was 0.94. This study suggests that the ERIC-PCR technique provides an alternative method for the identification of Listeria species and the discrimination of strains within one species.

En este estudio se analizaron 24 cepas de Listeria spp. De ellas, 22 fueron obtenidas en San Luis (Argentina), a partir de muestras humanas, de animales y alimentos. Se incluyeron 2 cepas de referencia Listeria monocytogenes CLIP 22762 y Listeria innocua CLIP 74915. Todos los aislamientos fueron identificados bioquímicamente y caracterizados por serotipificación, fagotipificación y detección del gen flaA por reacción en cadena de la polimerasa (PCR). Se generaron perfiles de bandas de ADN mediante la amplificación de secuencias repetitivas de consenso intergénico de enterobacterias (ERIC-PCR). De acuerdo a los resultados obtenidos por ERIC-PCR, las cepas de Listeria spp. fueron divididas en 3 grupos según su origen. Los perfiles de los aislamientos humanos y animales fueron distintos de los correspondientes a alimentos. Por otra parte, dentro de los grupos I y II se incluyeron 10 cepas de L. monocytogenes y solamente una de Listeria seeligeri. Dentro del grupo III no sólo estuvieron incluidas las 9 cepas de L. innocua sino también 4 de L. monocytogenes. La evaluación de los perfiles de bandas obtenidos por ERIC-PCR permitió la discriminación entre los serotipos ensayados 1/2b, 4b, 6a y 6b dentro de cada grupo. El índice de discriminación calculado para ERIC-PCR fue de 0,94. Los resultados sugieren que la técnica de ERIC-PCR provee un método alternativo válido para la identificación de especies de Listeria y, asimismo, permite la diferenciación de cepas dentro de una misma especie.

The role of surgical treatment in colon diverticulitis: indications and results.

PURPOSE: Colon diverticulitis is a common illness with affects 37-45% of western populations. Indications regarding therapy guidelines, operative timing and which surgical procedure to perform are still controversial. MATERIAL AND METHOD: Between January 1977 and December 1997, 239 patients, diagnosed with diverticulitis, have been admitted, on emergency, to our Department of General Surgery; 135 males (56%) and 104 females (44%), (mean age of 63 years). RESULTS AND DISCUSSION: Forty-two patients (18%), clearly diagnosed with diffuse or local peritonitis, underwent delayed emergency surgical procedure; 44 (22%) out of 197 patients, treated with medical therapy and subsequently underwent elective surgery procedures for complications (fistulas or stenosis). Among the 42 patients treated in emergency, 26 cases (62%) underwent to resection with immediate reconstruction. Among the elective surgery group 39 (89%) out of 44 underwent to resection with immediate reconstruction. Complications reached 40% in the group of emergency patients (mortality rate 12%) and 16% in the elective surgery group (mortality rate 2%). Several features possible influencing mortality rate have been analysed; age > 70 years, acute associated diseases, generalised peritonitis and surgical timing show a statistical significance. CONCLUSION: Therefore, a careful evaluation of the patients, an appropriate pre and post-operative medical treatment, with a wider use of the most recent techniques such as CT scan guided drain, intra-operative wash-out and peritoneal lavage are recommended in order to reduce morbidity and mortality.

[Helicobacter pylori infection diagnosis in dyspeptic patients from the City of San Luis, Argentina, using invasive methods]. / Diagnóstico de infección por Helicobacter pylori en pacientes dispépticos de la Ciudad de San Luis, Argentina, utilizando métodos invasivos.

In this study we present a relationship between different gastroduodenal pathologies and Helicobacter pylori infection. We used four diagnosis invasive methods for H. pylori infection: urease test (UT), histopathology (H), Gram stain (G) and culture (C). The upper gastrointestinal endoscopy of 300 dyspeptic patients showed that 71.6% had erosive congestive gastropathies, 13.6% had duodenopathies, 5.6% had gastric ulcer, 6.3% had duodenal ulcer and 2.6% had probable gastric neoplasia. We also correlated the data of water intake source with the pathologies. The percentage of infected patients with H. pylori was determined using: a) two simultaneous reference tests (UT and H), 54.3%, b) each test UT = 55.0%, H = 59.0%, G = 51.3%, and C = 43.0%. Sex, age and the source of water ingested did not show statistically significant differences.

Diagnóstico de infección por Helicobacter pylori en pacientes dispépticos de la Ciudad de San Luis, Argentina, utilizando métodos invasivos / Helicobacter pylori infection diagnosis in dyspeptic patients from the City of San Luis, Argentina, using invasive methods

In this study we present a relationship between different gastroduodenal pathologies and Helicobacter pylori infection. We used four diagnosis invasive methods for H. pylori infection: urease test (UT), histopathology (H), Gram stain (G) and culture (C). The upper gastrointestinal endoscopy of 300 dyspeptic patients showed that 71.6 had erosive congestive gastropathies, 13.6 had duodenopathies, 5.6 had gastric ulcer, 6.3 had duodenal ulcer and 2.6 had probable gastric neoplasia. We also correlated the data of water intake source with the pathologies. The percentage of infected patients with H. pylori was determined using: a) two simultaneous reference tests (UT and H), 54.3, b) each test UT = 55.0, H = 59.0, G = 51.3, and C = 43.0. Sex, age and the source of water ingested did not show statistically significant differences.

Diagnóstico de infección por Helicobacter pylori en pacientes dispépticos de la Ciudad de San Luis, Argentina, utilizando métodos invasivos / Helicobacter pylori infection diagnosis in dyspeptic patients from the City of San Luis, Argentina, using invasive methods

In this study we present a relationship between different gastroduodenal pathologies and Helicobacter pylori infection. We used four diagnosis invasive methods for H. pylori infection: urease test (UT), histopathology (H), Gram stain (G) and culture (C). The upper gastrointestinal endoscopy of 300 dyspeptic patients showed that 71.6 had erosive congestive gastropathies, 13.6 had duodenopathies, 5.6 had gastric ulcer, 6.3 had duodenal ulcer and 2.6 had probable gastric neoplasia. We also correlated the data of water intake source with the pathologies. The percentage of infected patients with H. pylori was determined using: a) two simultaneous reference tests (UT and H), 54.3, b) each test UT = 55.0, H = 59.0, G = 51.3, and C = 43.0. Sex, age and the source of water ingested did not show statistically significant differences.(AU)

Diagnóstico de infección por Helicobacter pylori en pacientes dispépticos de la Ciudad de San Luis, Argentina, utilizando métodos invasivos. / [Helicobacter pylori infection diagnosis in dyspeptic patients from the City of San Luis, Argentina, using invasive methods]

In this study we present a relationship between different gastroduodenal pathologies and Helicobacter pylori infection. We used four diagnosis invasive methods for H. pylori infection: urease test (UT), histopathology (H), Gram stain (G) and culture (C). The upper gastrointestinal endoscopy of 300 dyspeptic patients showed that 71.6

had erosive congestive gastropathies, 13.6

had duodenopathies, 5.6

had gastric ulcer, 6.3

had duodenal ulcer and 2.6

had probable gastric neoplasia. We also correlated the data of water intake source with the pathologies. The percentage of infected patients with H. pylori was determined using: a) two simultaneous reference tests (UT and H), 54.3

, b) each test UT = 55.0

, H = 59.0

, G = 51.3

, and C = 43.0

. Sex, age and the source of water ingested did not show statistically significant differences.

[Isolation of Helicobacter pylori from dental plaque]. / Aislamiento de Helicobacter pylori en placa dental.

It has been suggested that oral dissemination might be the major transmission vehicle for Helicobacter pylori, and that dental plaque might act as its reservoir. The presence of H. pylori was investigated in 62 odontological male and female patients (average age: 35 years old). Samples were taken from supragingival plaque, placed in 0.3 ml of thioglycolate broth, cultured within 12 h in Mueller-Hinton agar with the addition of 5-7% of sheep blood and antibiotic supplement, and incubated at 37 degrees C in microaerophilia for 5-7 days. Typical colonies were identified by gram, urease, oxidase and catalase. H. pylori was detected in a 15 year-old patient suffering from gastric acidity (1.61% positivity index). The medium used facilitated recovery of the agent from a sample abundant in germs. H. pylori was not recovered from the same patient 12 months later, suggesting that there might have been a transitory passage by gastric reflux or that the bacterium was acquired from an exogenous source.

Aislamiento de Helicobacter pylori en placa dental / Isolation of Helicobacter pylori in dental plaque

Se ha sugerido que la diseminación oral sería la principal vía de transmisión de Helicobacter pylori y la placa dental podría actuar como reservorio. Se investigó su presencia en 62 pacientes odontológicos de ambos sexos, edad promedio 35 años. Las muestras, tomadas de placas supragingival, colocadas en 0,3 ml de caldo tioglicolato, fueron cultivadas dentro de las 12 h en agar Mueller-Hinton con 5-7 por ciento de sangre de oveja y suplemento antibiótico e incubadas a 37ºC en microaerofilia 5-7 días. Las colonias típicas se identificaron por gram, ureasa, oxidasa y catalasa. H. pylori fue detectado en un paciente de 15 años que padecía acidez gástrica (índice de positividad 1,61 por ciento). El medio utilizado facilitó la