Lippia origanoides essential oil: an efficient and safe alternative to preserve food, cosmetic and pharmaceutical products.

AIMS: The aim of this work was to evaluate the efficacy and safety of Lippia origanoides essential oil as a preservative in industrial products. METHODS AND RESULTS: The composition, antimicrobial activity, mutagenic and toxic potential of L. origanoides were determined. Then, the effect of essential oil as a preservative in food, cosmetics and pharmaceutical products was evaluated. The essential oil of L. origanoides consisted mainly of oxygenated monoterpenes (38·13%); 26·28% corresponded to the compound carvacrol. At concentrations ranging from 0·312 to 1·25 µl ml-1 and in association with polysorbate 80, the essential oil of L. origanoides inhibited the growth of all the tested micro-organisms. The medium lethal dose in mice was 3·5 g kg-1 , which categorizes it as nontoxic according to the European Union criteria, and negative results in the Ames test indicated that this oil was not mutagenic. In combination with polysorbate 80, the essential oil exerted preservative action on orange juice, cosmetic and pharmaceutical compositions, especially in the case of aqueous-based products. CONCLUSIONS: Lippia origanoides essential oil is an effective and safe preservative for orange juice, pharmaceutical and cosmetic products. SIGNIFICANCE AND IMPACT OF THE STUDY: This study allowed for the complete understanding of the antimicrobial action and toxicological potential of L. origanoides essential oil. These results facilitate the development of a preservative system based on L. origanoides essential oil.

Evaluation of a multi-antigen test based on B-cell epitope peptides for the serodiagnosis of pulmonary tuberculosis.

SETTING: Two sample panels: 1) 20 pulmonary tuberculosis (PTB) patients and 10 healthy subjects from a country with a low incidence of TB (Italy); and 2) 47 PTB patients and 26 healthy subjects from a country with a high incidence of TB (Morocco). OBJECTIVE: To identify a combination of Mycobacterium tuberculosis peptides useful for the serodiagnosis of active PTB. METHODS: Fifty-seven B-cell epitope peptides of M. tuberculosis were evaluated by immunoenzymatic assay and the data were analysed using logistic regression analysis and the random forest method. RESULTS: The best discriminating peptide between PTB patients and healthy subjects from the sample of the low TB incidence country was the 23 amino acid peptide of the Rv3878 protein. The sensitivity and specificity were respectively 65% and 100%. The same peptide had a sensitivity and specificity of respectively 47% and 100% for the sample from the high TB incidence country. The best combination of peptides was a pool of nine peptides which had a sensitivity of 70.2% and a specificity of 100% in the high TB incidence country. CONCLUSIONS: The 9-peptide pool can be useful in identifying patients with active PTB.

M. avium binding to HLA-DR expressed alleles in silico: a model of phenotypic susceptibility to sarcoidosis.

Sarcoidosis is a systemic granulomatosis disease of unknown origin where a number of microbes, in particular M. tuberculosis and non-tuberculous mycobacteria, have been hypothesized to play a role in disease pathogenesis, possibly through bacterial antigen-driven hypersensitivity. To test this concept, we used bioinformatic tools allowing the identification of antigenic peptides in whole microbial genomes to analyze the interaction between the expressed HLA-DR gene allelic variants and the HLA-DR immunome of all pathogenic bacteria in a population of 149 sarcoidosis affected subjects and 447 controls, all HLA-typed at high resolution. We show here that patients with the Löfgren's syndrome, express HLA-DR alleles that recognize in silico a significantly higher number of bacterial antigen epitopes compared to the control population (18,496+9,114 vs 17,954+8,742; p<0.00001), and the chronic sarcoidosis affected population (17,954+8,742; p<0.00001 vs Löfgren's and controls). Further, the analysis of the ability of the HLA-DR allele combinations expressed by the Löfgren's and the chronic sarcoidosis affected subjects to recognize M. avium epitopes demonstrates that a significantly larger number of Löfgren's are capable of top affinity recognition, compared to chronic sarcoidosis (45% vs 17%, p<0.0037). Finally, both Löfgren's and chronic sarcoidosis subjects expressed HLA-DR allele combinations capable of M. tuberculosis and M. avium epitope recognition at higher affinity than tuberculosis affected subjects (p<0.01 all comparisons). In conclusion, we propose that - at least in a subgroup of affected subjects - sarcoidosis might be part of a spectrum of granulomatous responses to several agents where the Löfgren's syndrome represents the hyper-reactive end of the spectrum while pulmonary tuberculosis and atypical mycobacterial infections might represent the opposite end.

A model of phenotypic susceptibility to tuberculosis: deficient in silico selection of Mycobacterium tuberculosis epitopes by HLA alleles.

HLA-DR allelic variants have been associated with tuberculosis (TB) susceptibility in different populations with risk ratios of 3.7 to 7.2. We hypothesized that the genetic susceptibility to TB depends upon the reduced capability of HLA-class II alleles of TB patients to bind and select peptide antigen from the Mycobacterium tuberculosis (MTB) expressed genome. To test this hypothesis, we developed a software that can predict HLA-DR restricted epitopes within the whole MTB genome based on quantitative peptide binding matrices. We analyzed the number of MTB epitopes recognized in two previously described populations of TB patients and matched controls and in a control population comprised of individuals affected by a sarcoid-like granuloma induced by beryllium and by healthy exposed controls. The number of putative epitopes within the whole MTB genome which could be bound by any HLA-DR allele (HLA-DR immunome of MTB) was 405,422 out of 1,304,277 possible 9-mers i.e., 31.08% of the global capability, instead of the expected 35%. When tested at an affinity level equivalent of the 1% of the best binder peptides, the HLA-DR alleles (HLA-DRB1*0801, *0802, *1401, *1501 and *1502) associated with TB susceptibility recognized a significantly lower mean number of MTB-epitopes (7,862 +/- 4,258) than the MTB-epitopes recognized by HLA-DR alleles (HLA-DRB1*0301, *0701, *1101, *1102, *1301 and *1302) negatively associated with TB (11,376 +/- 1,984, p<0.032). The number of epitopes bound at high affinity out of the whole MTB genome by the combination of the two HLA-DR alleles carried by each individual was lower in TB patients [TB-population 1: 11,341 +/- 908 (mean+SEM); TB-population 2: 15,303 +/- 657] than in matched healthy controls (CTR-population 1: 13,587 +/- 605, p<0.03 vs TB-population 1; CTR-population 2: 1,6841 +/- 555, p<0.04 vs TB-population 2). No difference was seen in individuals with the sarcoid-like granuloma induced by beryllium compared to the exposed healthy (beryllium-hypersensitivity: 17,593 +/- 447; controls 18,014 +/- 421; p=0.57). The data suggest that HLA-DR alleles associated with susceptibility to tuberculosis may be endowed with a reduced capability to bind at high affinity T-cell epitopes and select them for antigen presentation. The same alleles may contribute to determine the reaction to mycobacteria in non tuberculous granulomatous disorders.

Routine laparoscopic cholecystectomy after endoscopic sphincterotomy for choledocholithiasis in octogenarians: is it worth the risk?

BACKGROUND: No unanimous consensus has been reached as to the need for routine laparoscopic cholecystectomy (LC) after endoscopic sphincterotomy (ES) for choledocholithiasis in very elderly patients, who are considered as high-risk subjects for surgery. METHODS: From 1991 through 1997, 170 patients were referred to undergo preoperative ES and routine LC for common bile duct (CBD) stones. The results for 27 patients (age 80 years or older) were compared with those achieved for younger patients. Successively, in a retrospective case-control study, the results for the selected patients were compared with those for 27 very elderly patients who underwent endoscopic retrograde cholangiopancreatography (ERCP), but did not receive LC. The mean follow-up period was 126 months. RESULTS: Octogenarians showed longer surgery time (79 vs 51 min) and postoperative hospital stay (2.8 vs 1.2 days), as well as more early low-grade complications (15% vs 3%), whereas there were no differences in conversion rate or serious complications. Recurrent symptoms or complications developed in 48% of octogenarians not undergoing routine LC, and 30% finally needed surgery. One patient in the control group died after emergency cholecystectomy for acute cholecystitis. The results of surgery were significantly poorer for the control group. CONCLUSIONS: Although a "wait-and-see" policy allowed two-thirds of LCs to be avoided in octogenarians, biliary-related events developed for every second patient, often requiring delayed surgery, with poorer results. Sequential treatment (ES followed by elective LC) is a safe procedure for octogenarians, and should be considered as a standard, definitive treatment for cholecystocholedocholithiasis even after the age of 80 years.

Evidence based medicine and effective interventions of pharmacological therapy for the prevention of osteoporotic fractures.

Osteoporosis and fractures associated with it constitute a real and serious socio-medical problem, which only recently has come to the forefront of social consciousness. The authors are carrying out a critical re-examination of the medical literature of osteoporosis pharmacological treatment. Particular attention has been paid to studies which show a clear reduction of the primary endpoint that, in the case of this pathology, consists of a reduction of the fracturing event. According to the examination of the clinical studies introduced, antiresorptive bone agents, such as alendronate and risenderonate, turn out to be molecules with higher levels of evidence implicated on the reduction of the main osteoporotic fractures, in particular the reduction of vertebral and femoral fractures. The 10 years long-term extension studies, in particular those that have seen the employment of alendronate, found a positive outcome regarding densitometry data and a favorable trend in antifracture effectiveness. Ibandronate is another amino-bisphosphonate which was recently validated as an effective drug for the treatment of osteoporosis with its documented ability to meaningfully reduce vertebral fractures. Also ranelate of strontium, a drug that seems to explain its own result in a different way from the other antiresorptive bone agents, constitutes another valid alternative in the treatment of this pathology. Both of these molecules however, need further studies in order to estimate their antifracture effectiveness in the long term, particularly those related to femoral fractures. Teriparatide and the entire molecule paratohormone are usually not prescribed for its high cost in treatment and because, typically, patients with high-risk level fractures that are already affected, produce more vertebral fractures from moderate to severe intensity.

The application of positive matrix factorization in the analysis, characterisation and detection of contaminated soils.

Multivariate factor analytical techniques are widely used for the approximation, in terms of a linear combination of factors, of multivariate experimental data. The chemical composition of soil samples are multivariate in nature and provide datasets suitable for the application of these statistical techniques. Recent developments of multivariate factor analytical techniques have led to the approach of Positive Matrix Factorization (PMF), a weighted least squares fit of a data matrix in which the weights are determined depending on the error estimates of each individual data value. This approach relies on more physically significant assumptions than methods like Principal Components Analysis which is frequently used in the analysis of soil datasets. In this paper we apply PMF to characterise the pollutant source in a set of geographically referenced soil samples taken within a 200 m radius of a site characterised by a high concentration of heavy metals. Each sample has been analysed for major and minor elements (using wavelength-dispersive X-ray fluorescence spectrometry), carbon, hydrogen and nitrogen (using a CHN elemental analyzer) and mercury (using cold-vapour atomic absorption spectrometry). Analysis of the soils using PMF resulted in a successful partitioning of variances into sources related to background soil geochemistry, organic influences and those associated with the contamination. Combining these results with a geostatistical approach successfully demarcated the main source of the combined organic and heavy metal contamination.

Dysphagia aortica: a neglected symptom of aortoesophageal fistula.

Aortoesophageal fistula, secondary to thoracic aortic aneurysm, is an uncommon cause of gastrointestinal bleeding that is uniformly fatal without surgical intervention. Typical symptoms are midthoracic pain and/or dysphagia followed by a usually short, albeit unpredictable, symptom-free interval and by a 'herald' haemorrhage, which is observed in 80% of patients before fatal exsanguinations. Dysphagia is present in 45% of patients, sometimes for several weeks, before the first bleeding occurs. However, dysphagia aortica is rarely considered in the differential diagnosis of dysphagia and lack of awareness, as well as symptom's underevaluation, both contribute to a significant diagnostic and therapeutic delay. We present a case of a 77-year-old woman who died for a bleeding AEF consequent to a thoracic aortic aneurysm and whose main symptom during the past 2 months was dysphagia, which was not taken seriously into consideration by her general practitioner. This case report emphasises that primary care physicians should be alerted to evaluate carefully the alarming symptoms like dysphagia -- especially in elderly patients -- before life threatening complications occur, as they are the ones who could suspect early the diagnosis and make a proper referral.

Perioperative management of diabetic subjects. Subcutaneous versus intravenous insulin administration during glucose-potassium infusion.

The purpose of this study was to validate methods for the perioperative management of diabetic patients that meet the prerequisites of simplicity, applicability in the absence of a diabetologist, and flexibility, to rapidly meet changing metabolic requirements. The patients were divided into two groups that were comparable for age, sex distribution, type of diabetes, and type of surgical procedures. The results show that intravenous insulin administration achieved better glycemic control during the intraoperative period, whereas it did not offer advantages over the subcutaneous route during the pre- and postoperative periods. The satisfactory degree of steady glycemic control achieved and the absence of hypoglycemic episodes indicate that the separate administration of insulin and glucose plus electrolytes is an effective and safe management modality for diabetic patients undergoing major surgery.

Does laparoscopic cholecystectomy worsen the prognosis of unsuspected gallbladder cancer?

BACKGROUND: Several reports claim that there is a risk that laparoscopic cholecystectomy (LC) might worsen the prognosis of unsuspected gallbladder cancer. HYPOTHESIS: Several factors rather than LC could influence prognosis. METHODS: A retrospective clinicopathologic study was performed on 20 patients, 9 patients (3 men and 6 women, aged from 36 to 75 years [mean age, 62.3 years]) undergoing LC and 11 patients (2 men and 9 women, aged from 53 to 91 years [mean age, 65.3 years]) undergoing open cholecystectomy (OC), with postoperatively diagnosed gallbladder cancer. The correlation was evaluated between cumulative survival rates and the following 7 prognostic factors: age, sex, histopathological grade, pathologic stage, occurrence of bile spillage, type of cholecystectomy (LC or OC), and additional surgical treatments. RESULTS: Seven patients (87%) after LC and 9 patients (82%) after OC had cancer recurrence: the difference is of no statistical significance (P =.9). There were no recurrences of cancer in the abdominal wall after either LC or OC. Survival rate was statistically correlated to tumor stage (P =.007) and to the occurrence of bile spillage (P =.002). Survival rate did not change according to whether the operation was carried out using LC or OC (P =.60). CONCLUSION: These results would seem to lend support to the opinion that LC does not worsen the prognosis for unsuspected gallbladder cancer.

In vitro evaluation of the cytotoxic, hemolytic and clastogenic activities of Rhizostoma pulmo toxin(s).

Cytotoxic, hemolytic and clastogenic activities of Rhizostoma pulmo toxin(s) contained in the jelly tissue free of nematocysts were investigated in mammalian cells with in vitro procedures. At the concentration of 37.6 microg/ml the tissue protein produced the death of 50% V79 cells; a similar potency was observed in terms of hemolytic activity. The toxin(s) was not clastogenic for human lymphocytes in culture at the concentration of 5 microg/ml.

Endoscopic treatment of gastro-oesophageal reflux disease (GORD): a systematic review.

Gastro-oesophageal reflux disease represents an extremely common disorder which has a substantial impact on patients' quality of life and use of health care resources. Gastro-oesophageal reflux disease is a chronic relapsing disease for which a lifelong solution is needed. Until now the two competing therapeutic modalities have been the medical and surgical therapies. Quite recently a third option has become available. A number of endoscopic anti-reflux procedures have been described, with the common goal of creating an anti-reflux barrier, thus obviating long-term proton pump inhibitors and the cost and potential risk of laparoscopic Nissen fundoplication. In this review the different techniques are thoroughly examined and the results are critically evaluated, giving special emphasis to efficacy, safety and durability of these new anti-reflux procedures. Available data show that these anti-reflux techniques produce significant improvement in gastro-oesophageal reflux disease symptomatology and quality of life as well as reduce the use of anti-reflux medication, without causing serious morbidity or mortality. However, the majority of these techniques have failed to adequately control oesophageal acid reflux. Endoscopic anti-reflux therapies therefore sound very attractive-being less invasive than surgery-and show a significant promise, but are still in the early stages of assessment. Large-scale randomized multi-centre trials comparing control groups with sham procedures are essential to confirm their efficacy. Further studies are also necessary to determine what modifications these techniques require in order to produce maximum clinical efficacy and durability. However, considering that current therapies (both medical and surgical) of gastro-oesophageal reflux disease are highly effective, the need for such new endoscopic modalities may be questionable. Moreover, appropriate trials in dedicated centres should be carried out to assure that the enthusiasm commonly associated with new technology is justified and can be generalized to open-access endoscopists.

Reliability of ultrasounds in detecting common bile duct stones before biliary surgery. A retrospective analysis.

UNLABELLED: The aim of this study is to evaluate the diagnostic benefit of US in the preoperative screening for CBD stones in patients submitted to surgery for biliary lithiasis. MATERIALS AND METHODS: We analyzed the data of 135 patients admitted for biliary lithiasis, jaundice or biliary pancreatitis in the last three years. Mean age was 63 years (54.8% females), 48% of the cases had an urgent admission. US of the liver and biliary ducts was the first examination performed and the patients were divided in 2 groups on the bases of the results: those with the diameter of the CBD < 1 cm (101 pts-group A) and those with a diameter > 1 cm (34 pts-group B). RESULTS: In group A were subsequently performed: 35 IV cholangiographies (6 showed CBD stones, 1 was a false negative for CBD stones), 15 intraoperative (I.O.) cholangiographies (1 positive for CBD stones), 9 ERCP for jaundice or pancreatitis (2 positive for CBD stones). The incidence of CBD stones was 8.9% in this group. In group B were performed: 7 IV cholangiographies (2 showed CBD stones), 12 I.O. cholangiographies (6 showed CBD stones), 10 ERCP for jaundice or pancreatitis (7 showed CBD stones). The incidence of CBD stones was 44% in this group. CBD stones were detected by US in 2 cases (22%) in group A and in 6 cases (40%) in group B. CONCLUSIONS: In our patients there is a relatively high incidence of CBD stones (8.9%) in spite of a normal CBD at US. Considering that the minimally invasive approach for the gallbladder stones is becoming the "gold standard" and the laparoscopic treatment of the CBD stones is not widely diffused, it is important to know the presence of CBD stones before surgery. With this in mind US does not seem to be a reliable diagnostic tool and we suggest that before surgery an IV cholangiography (sensitivity > 95% in our experience) should be performed together with US and ERCP in selected cases. Moreover this could reduce the number of preoperative ERCP leaving the possibility to treat more selectively by endoscopic papillosphincterotomy the patients with CBD stones before laparoscopic cholecystectomy.

Hand-assisted laparoscopic splenectomy for a splenic hamartoma.

We present a patient with a hamartoma of the spleen. The case was thought to be a good indication for a hand-assisted laparoscopic approach and treatment was successful. Until now, only one case of a laparoscopic removal of a splenic hamartoma has been reported. Focal lesions of the spleen should be removed intact to allow a complete histological examination and to avoid peritoneal dissemination in case of malignancy. In these patients, the hand-assisted approach makes possible the removal of an unminced organ and has the advantage of being a purely laparoscopic technique. It should thus be considered the method of choice for surgical treatment.

Does the learning phase influence the late outcome of patients with gastroesophageal reflux disease after laparoscopic fundoplication?

BACKGROUND: Although the degree of surgical experience clearly affects early outcome of laparoscopic antireflux surgery, its influence on long-term results has not been fully evaluated. The aim of this study was to verify whether the initial experience in laparoscopic antireflux surgery could also influence the late clinical outcome. METHODS: Clinical and endoscopic findings, together with quality of life, of the first 25 patients successfully submitted to laparoscopic fundoplication were compared with those of 25 matched controls operated on later. RESULTS: At more than 2 years', follow-up, reflux symptoms, endoscopic findings, use of antisecretory drugs, side effects, and quality of life were not significantly different in both groups, despite a high occurrence of major anatomical failures (three vs one) in the first set of patients. CONCLUSION: The late clinical outcome of patients with gastroesophageal disease operated on during the learning phase or after gaining experience is not different, provided the surgeon is adequately trained in laparoscopic surgery.

Gallbladder injury in blunt abdominal trauma.

A 36-year-old woman was admitted to the hospital for an abdominal blunt trauma. At ultrasound (US) and computed tomography (CT), a gallbladder lesion was suspected, along with a tear of the liver. The patient was submitted to a diagnostic laparoscopy. The gallbladder was partially avulsed and bile was infiltrated in the hepatoduodenal ligament. Intraoperative cholangiography and Kocher's maneuver excluded other lesions. Laparoscopic cholecystectomy was performed, but due to severe hemorrhage from the liver tear, the operation was converted to an open procedure. Gallbladder lesions in blunt trauma are rare occurrences, but they are often associated with other organ injuries. US and CT scan are valuable for their diagnosis, but if a lesion is suspected, diagnostic laparoscopy is advisable in stable patients. It should be accompanied by a cystic duct cholangiography and a Kocher's maneuver to evaluate the integrity of the biliary tree. Laparoscopic cholecystectomy is generally feasible. Associated lesions require laparotomy when they are not amenable to laparoscopic treatment.

Quality of life for patients with gastroesophageal reflux disease 2 years after laparoscopic fundoplication. Evaluation of the results obtained during the initial experience.

BACKGROUND: Health-related quality of life (HRQL) is significantly affected by gastroesophageal reflux disease (GERD), and its evaluation is emerging as a factor important to select treatment options for GERD. Antireflux laparoscopic surgery improves HRQL. The aim of this study was to compare the preoperative and medium-term postoperative HRQL in patients submitted to laparoscopic fundoplication during the initial experience to verify its possible influence on HRQL outcomes. METHODS: Clinical assessment, endoscopy, and a previously validated HRQL index were performed before and 2 years after surgery in 32 patients who underwent at the beginning of our experience. RESULTS: The esophageal mucosa returned to normal 2 years after laparoscopic fundoplication in 81% of the patients. Heartburn was absent or occasional in 93%, and 65% were free of antisecretory drugs. All postoperative HRQL items were significantly improved (p <0.0001), with the postoperative curve of HRQL scores superposable to those of healthy Italian subjects (Italian normative sample). CONCLUSIONS: The initial phase of learning does not affect the improvement of HRQL observed after laparoscopic antireflux surgery, which is consistent with durable relief of symptoms and endoscopic healing. Evaluation of HRQL should be added to, and probably could replace in most cases, the objective postoperative testing.

Abuse and therapeutic potential of gamma-hydroxybutyric acid.

Gamma-hydroxbutyric acid is a compound found in mammalian brain that is structurally related to the neurotransmitters gamma-aminobutyric acid and glutamic acid. Gamma-hydroxybutyric acid effects dopaminergic systems in the brain and may be a neurotransmitter. Gamma-hydroxybutyric acid was first reported as a drug of abuse in 1990 and continues to be abused by bodybuilders, participants of "rave" dance parties, and polydrug abusers. Physical dependence can develop after prolonged, high-dose use, and overdoses have been widely reported. Its use in sexual assaults as a "date rape" drug and availability on the internet have recently emerged. Gamma-hydroxybutyric acid has established efficacy as an anesthetic agent, and preliminary evidence supports its utility in the treatment of alcohol dependence, opiate dependence, and narcolepsy.

Abdominal aortic aneurysms: some controversial points.

A retrospective review of the surgical treatment of 51 abdominal aortic aneurysms is reported. Thirty-five (69%) patients were operated on electively, and 16 (31%) had emergency surgery. Fissurated aneurysms were included in the elective surgery group. The operative death rate was respectively 2.7% and 50%. Controversial points about diagnosis, treatment of associated diseases, surgical technique, and selection of the patients for surgery are presented and discussed.

Surgical treatment of left main coronary stenosis. Notes on 179 consecutive patients.

The authors report the clinical haemodynamic and angiographic data and early and late postoperative results of 179 patients consecutively operated on for left main coronary artery stenosis. Operative mortality was 3.9%; incremental risk factors were identified by univariate analysis and included female sex, unstable angina pectoris and congestive heart failure. Late postoperative mortality was 4.9%. After a mean follow-up of 31 months overall survival is 91.6%; the great majority of survivors are doing well, free from anginal pain, while 16.4% of patients experience recurrent angina pectoris of lesser severity than before operation (I-II C.C.S. class) and well controlled by medical treatment alone.