Correction to: Increasing the synthesis of bioactive abietane diterpenes in Salvia sclarea hairy roots by elicited transcriptional reprogramming.

Unfortunately, the second author name was wrongly published in the original publication. The correct author name should read as follows.

Increasing the synthesis of bioactive abietane diterpenes in Salvia sclarea hairy roots by elicited transcriptional reprogramming.

KEY MESSAGE: Transcriptional activation of genes belonging to the plastidial MEP-derived isoprenoid pathway by elicitation with methyl jasmonate and coronatine enhanced the content of bioactive abietane diterpenes in Salvia sclarea hairy roots. We have shown that aethiopinone, an abietane diterpene synthesized in Salvia sclarea roots is cytotoxic and induces apoptosis in human melanoma cells. To develop a production platform for this compound and other abietane diterpenes, hairy root technology was combined with the elicitation of methyl jasmonate (MeJA) or the phytotoxin coronatine (Cor). Both MeJA and Cor induced a significant accumulation of aethiopinone, but prolonged exposure to MeJA irremediably caused inhibition of hairy root growth, which was unaffected by Cor treatment. Considering together the fold increase in aethiopinone content and the final hairy root biomass, the best combination was a Cor treatment for 28 days, which allowed to obtain up to 105.34 ± 2.30 mg L-1 of this compound to be obtained, corresponding to a 24-fold increase above the basal content in untreated hairy roots. MeJA or Cor elicitation also enhanced the synthesis of other bioactive abietane-quinone diterpenes. The elicitor-dependent steering effect was due to a coordinated transcriptional activation of several biosynthetic genes belonging to the plastidial MEP-derived isoprenoid pathway. High correlations between aethiopinone content and MeJA or Cor-elicited level of gene transcripts were found for DXS2 (r 2 = 0.99), DXR (r 2 = 0.99), and GGPPS (r 2 = 0.98), encoding enzymes acting upstream of GGPP, the common precursor of diterpenes and other plastidial-derived terpenes, as well as CPPS (r 2 = 0.99), encoding the enzyme involved in the first cyclization steps leading to copalyl-diphosphate, the precursor of abietane-like diterpenes. These results point to these genes as possible targets of metabolic engineering approaches to establish a more efficient production platform for such promising anti-proliferative plant-derived compounds.

Eugenol and thymol, alone or in combination, induce morphological alterations in the envelope of Candida albicans.

The envelope of Candida albicans, with its outermost array of macromolecules protruding towards the environment, is pivotal to the expression of major virulence factors such as adhesiveness, and the morphological transition to hyphal form. We tested the anticandidal activity of eugenol, main component of clove oil, and thymol, main component of thyme oil, alone or in combination, by investigating their ability to interfere with the architecture of the envelope of C. albicans. Both molecules alterated the morphogenesis of the envelope, but the effects of thymol were more pronounced than those of eugenol. Certain combinations of the two molecules led to a synergistic effect, which is interesting in the view of potentiating their inhibition of C. albicans colonisation and infectiousness.

A comparison of fat intake of normal weight, moderately obese and severely obese subjects.

BACKGROUND: Excess dietary fat has been implicated in the etiology of obesity. METHODS: This study examined the fat intake of three weight groups, normal (20.0 < or = BMI < or = 27.0), moderately obese (27.1 < or = BMI < or = 39.9) and severely obese (BMI > or = 40.0). Each group contained 50 subjects. Detailed 3-day food records were used to gather the nutritional data. Anthropometric and sociodemographic information was also collected. RESULTS: Overall fat intake was 89 +/- 42 g/day or 37 +/- 10% of total energy. Total fat (g/1000 kcalories) intake was found to be significantly higher in the obese groups (p < 0.05). Subjects in the moderately and severely obese groups consumed significantly more fat and cholesterol and less carbohydrate than did normal weight subjects. Compared to the normal weight subjects, obese subjects also had higher intakes of saturated, monounsaturated and polyunsaturated fat (as a percentage of dietary energy). There was no difference in energy or protein intake, and P/S ratio among the three groups. BMI was strongly positively correlated with total fat, saturated, monounsaturated, polyunsaturated fat, cholesterol, and protein intake (as g/day only), and negatively correlated with carbohydrate intake and the CHO/FAT ratio. Energy intake was not significantly associated with BMI. CONCLUSION: A high fat diet may promote obesity, independently of its calorie contribution.

Abnormal trace elements in a patient on total parenteral nutrition with normal renal function.

OBJECTIVE: To describe trace metal changes in a 74-year-old male patient with mesenteric fibrosis and a small bowel fistula who was maintained on total parenteral nutrition in the hospital and at home. METHODS: Trace elements which included chromium and selenium were monitored over a 14-month period as part of his nutrient follow-up. RESULTS: Serum chromium reached levels > 21-fold the upper reference range, and serum selenium, in contrast, was < 0.5 the lower reference range. Plasma aluminum was also measured, and found to be nearly twice the upper reference range, although the patient had normal renal function. We measured the aluminum content of the parenteral nutrients and additives, and found that replacement of calcium gluconate by calcium chloride helped to reduce the aluminum content in the final parenteral solution by 34%. Aluminum and chromium contaminants found in parenteral solutions need to be reduced or removed to avoid toxic accumulation. CONCLUSION: This study illustrates the importance of adequately adjusting essential trace elements, and monitoring contaminants in parenteral fluids in an individual on total parenteral nutrition.

The In Situ Detection of PCR-Amplified Hepatitis C RNA.

The discovery of PCR in 1985 has had an enormous impact on the field of molecular diagnostics. However, an important drawback of solution-phase PCR is that one cannot localize the target of interest to a specific cell type owing to the obligatory tissue destruction required for DNA extraction. This problem has been circumvented by the field of in situ PCR. This chapter will describe the theoretical basis of in situ PCR using hepatitis C as a model system. The goals of this manuscript are: 1. To describe the various DNA synthesis pathways that may be operative during in situ PCR. 2. To describe the key preparatory steps of reverse transcriptase (RT) in situ PCR 3. To describe the actual protocol of RT in situ PCR 4. To discuss potential problems when doing RT in situ PCR and how to resolve these problems, using hepatitis C infection of liver tissues as the model system.

Evolving cooperation: the role of individual recognition.

To evaluate the role of individual recognition in the evolution of cooperation, we formulated and analyzed genetic algorithm model (EvCo) for playing the Iterated Prisoner's Dilemma (IPD) game. Strategies compete against each other during each generation, and successful strategies contribute more of their attributes to the next generation. Each strategy is encoded on a 'chromosome' that plays the IPD, responding to the sequences of most recent responses by the interacting individuals (chromosomes). The analysis reported in this paper considered different memory capabilities (one to five previous interactions), pairing continuities (pairs of individuals remain together for about one, two, five, or 1000 consecutive interactions), and types of individual recognition (recognition capability was maximal, nil, or allowed to evolve between these limits). Analysis of the results focused on the frequency of mutual cooperation in pairwise interactions (a good indicator of overall success in the IPD) and on the extent to which previous responses by the focal individual and its partner were associated with the partner's identity (individual recognition). Results indicated that a fixed, substantial amount of individual recognition could maintain high levels of mutual cooperation even at low pairing continuities, and a significant but limited capability for individual recognition evolved under selection. Recognition generally increased mutual cooperation more when the recent responses of individuals other than the current partner were ignored. Titrating recognition memory under selection using a fitness cost suggested that memory of the partner's previous responses was more valuable than memory of the focal's previous responses. The dynamics produced to date by EvCo are a step toward understanding the evolution of social networks, for which additional benefits associated with group interactions must be incorporated.

Cytologic correlates of benign versus dysplastic abnormal keratinization.

The purpose of this study was to determine the cytologic and histologic features that differentiate benign from squamous intraepithelial lesion (SIL)-associated cervical abnormal keratinization, defined as hyperkeratosis, parakeratosis, or individual cell dyskeratosis. Fifty-four cervical Papanicoloau (Pap) smears that contained abnormally keratinized cells were reviewed without knowledge of the concurrent biopsy. Twenty-three Pap smears were diagnosed as SIL and the corresponding biopsy showed SIL in 21 (91%) of these cases. Of the 23 Pap smears diagnosed as negative for SIL, the corresponding biopsy in 20 cases (87%) showed benign (SIL negative) abnormal keratinization. Eight Pap smears showed squamous atypia, of these 5 showed SIL on biopsy, and the other 3 revealed benign keratinization. The Pap smear correlates of the 25 biopsies that were negative for SIL included marked hyperkeratosis (18/25-72 vs. 5/29-17% for biopsies with SIL) and regular nuclear membranes (16/18-89% cases with nucleated dyskeratotic cells vs. 5/29-17% for biopsies with SIL). The cytologic correlates of the 29 biopsies that showed SIL included irregular chromatin clumping (27/29-93% vs. 3/18-17% for biopsies without SIL), and a disorganized growth pattern (24/29-83 vs. 5/25-20% for biopsies without SIL). It is concluded that the cytologic distinction between benign and SIL-related Pap smears with abnormal keratinization can be reliably made by the degree of hyperkeratosis, nuclear chromaticity pattern and contour, and the growth pattern of the dyskeratotic cells.

Can elevated chromium induce somatopsychic responses?

The possible somatopsychological effects of chromium (Cr) was investigated in a population of patients, from a surgical ward of our hospital, who required total parenteral nutrition (TPN) solutions, and who became exposed to various amounts of this metal from this treatment. The study involved a questionnaire as well as biochemical tests which included serum Cr and other selected trace metals. The renal status for all eligible patients was within normal parameters. The patient population varied in age, pathology, surgical treatment, and duration on TPN. The results showed that every patient who received TPN had an increased serum Cr level; some increases were up to 50-fold above the normal reference level for serum Cr. Although statistical analysis failed to show any significant statistical relationship between an increased serum Cr and the investigated somatopsychological disturbances, this effect cannot be ruled out since one case did show all the dream disturbances. Considering these cases, the action of sedative medications that may suppress the effects of Cr, cannot be ruled out. As Cr(III) may be potentially genotoxic at high concentrations, infusion of this metal over long time periods should be avoided. Supplementation of Cr in TPN solutions appears to be unnecessary for short-term TPN because this metal is a known contaminant of these solutions. Efforts are required to find TPN nutrients with low or no Cr contamination.

Selenium and zinc levels in surgical patients receiving total parenteral nutrition.

The zinc and selenium levels of 40 surgical patients were monitored pre- and post-TPN. The initial selenium level was low normal, and the initial zinc level was also low. Both selenium and zinc are potent antioxidants involved in cellular defense against free radicals. Surgical patients are at risk for selenium and zinc deficiencies secondary to both increased needs and losses. TPN blood work protocols should include monitoring of selenium and zinc with supplementation of the nutrient solutions, as required.

Classic and common migraine suggestive clinical evidence of two separate entities.

To help clarify whether or not classic migraine (C1M) and common migraine (CoM) should be considered as a single entity, we compared the clinical features of a group of C1M patients with those of a group of CoM patients. Significant differences between the two forms of migraine were found in the following areas: age distribution of sufferers; family history of migraine, ischemic cerebrovascular disease, and coronary heart disease; age at onset; frequency of attacks; duration, location, and severity of pain; accompanying symptoms; and, evolution during weekends, menstruation, pregnancy, and puerperium. Our comparative investigation suggests that CIM and CoM are more distinct than previously considered.

An integrated team approach to the management of patients with oropharyngeal dysphagia.

This paper describes a team approach to the assessment and management of patients with oropharyngeal dysphagia of neurologic origin. The team's major focus was to determine the need for adjustments to the patient's diet to maintain or restore the safety of oral feeding. This involved the development of a detailed radiographic examination and a series of dysphagia diets, in addition to comprehensive evaluations by an occupational therapist, physiotherapist, and speech pathologist. The effects of deteriorating swallowing ability on the physical, cognitive, and emotional status of the patient are discussed in the context of a multidisciplinary approach.

[Wet oxidation of toxic industrial waste with oxygenated water]. / L'ossidazione a umido con acqua ossigenata dei reflui tossici di origine industriale.

The industrial toxic waste streams hot treatment technology with hydrogen peroxide and catalysts, developed by the research laboratories of Montefluos in Bollate, allows the abatement of many organic and bio-toxic pollutants. Some treatment examples are here reported. The examples, performed on a laboratory scale, relate to industrial waste streams with a high COD (100000-200000 mg/l) in which it was possible to obtain an abatement over the 90% of pollutants like phenols, formaldehyde, dimethylformamide and phenyl acetate. The application range of this technology is similar to that of oxygen or air wet oxidation, but it has remarkable advantages due to the lower plant, maintenance and energy costs, because the treatment is performed using much more bland conditions (atmospheric pressure and 90-100 degrees C of temperature). The aim of the bio-toxic pollutants abatement and COD reduction (70-80%) is to allow the final bio-digestion waste streams with high organic content, but too diluted to be directly incenerated at a suitable cost.

Case mix at the European Institute of Oncology: first report of the Tumour Registry, 2000-2002.

INTRODUCTION: An institutional and centralized hospital-based tumour registry (TR) is the ideal supporting tool for the organization and management of clinical data in a comprehensive cancer centre. The purpose of this paper is to describe the development of the TR at the European Institute of Oncology (IEO), Milan, Italy, from its origin to its current applications. MATERIAL AND METHODS: After a series of meetings with members of administrative, clinical, research and informatics departments, the TR was activated in March 2006 with the aim of collecting data on all the individuals referred to the institute, with or at risk of developing a tumour. It was implemented on an Oracle™-based interface. A minimum dataset of variables was defined and data collection was divided into four forms, which together gather all the relevant data on patients, tumours, treatments and subsequent events. RESULTS: After a six-month pilot period, which involved the training of the tumour registrars, adjustments to the structure of the registry, development of a data quality control procedure and finalization of the operative protocol, since September 2006 the data collection has been fully operative. Five registrars have been chronologically entering data of all individuals who visited the IEO for the first time since 1 January 2000. As of March 2009, data on 69,637 individuals and 43,567 tumours has been reviewed, recoded and registered in the TR. Twenty-two per cent of the tumours (n=9578) were first invasive primaries, diagnosed and treated in the IEO; the most common sites were breast (n=4972), lung (n=627), intestines (n=479) and prostate (n=376). CONCLUSION: The IEO TR has been proven functional and reliable in monitoring the activity of the hospital, allowing extraction of data from any subpopulation with characteristics of interest. The structured and centralized TR represents an important tool for our research-oriented institution.

Inhibitory activity of thymol against the formation and viability of Candida albicans hyphae.

As the capacity of Candida albicans to produce hyphae is considered an important virulence factor in the pathogenesis of candiasis, the aim of this study was to investigate whether thymol, the major component of thyme oil, can interfere with the filamentous forms of Candida albicans and their viability. The morphological transition from yeasts to filamentous forms was investigated by analysing the morphological index (MI), which classifies the differentiated forms and blastoconidia; viability was investigated by means of fluorescence microscopy using a new SYTO-9 and propidium iodide method previously used to stain only blastoconidia. Without thymol, there was an average of 94.00 +/- 3.06% hyphal forms. After 6 h of incubation with 1x MIC (125 microg ml(-1)), 1/2x MIC and 1/4x MIC of thymol, filamentation was, respectively, 14.33 +/- 8.25%, 28.33 +/- 7.17% and 45.67 +/- 8.09% in comparison with control (all statistically significant). In the absence of thymol, viable cells accounted for an average of 93.00 +/- 4.00% whereas, after 6 h of incubation with 1x MIC, 1/2x MIC and 1/4x MIC of thymol, the presence of 54.33 +/- 1.86%, 29.00 +/- 3.61% and 23.00 +/- 2.52% of yellow-orange coloured forms indicated damaged membranes and reduced viability. Our findings show that thymol interferes with the formation and viability of hyphae. This can be attributed to the characteristics of thymol disturbing Candida cell membranes and metabolism, probably by affecting fungal cell-wall synthesising enzymes.

AIDS dementia is associated with massive, activated HIV-1 infection and concomitant expression of several cytokines.

BACKGROUND: We recently showed that acquired immunodeficiency syndrome (AIDS) dementia is associated with activated infection of microglia, neurons, and astrocytes by HIV-1. However, it is doubtful whether infection per se is responsible for the dramatic symptoms associated with AIDS dementia. The purpose of this study was to determine the histologic distribution of messenger RNAs (mRNAs) of several cytokines that have been implicated in AIDS pathogenesis and to correlate this expression pattern with the in situ localization of polymerase chain reaction (PCR)-amplified HIV-1 nucleic acids in the central nervous system (CNS). MATERIALS AND METHODS: HIV-1 DNA was detected by PCR in situ hybridization. HIV-1 RNA and cytokine expression, including tumor necrosis factor alpha (TNF), inducible nitric oxide synthetase (iNOS), and macrophage inflammatory protein alpha (MIP-1 alpha) and MIP-1 beta mRNA were detected by reverse transcriptase (RT) in situ PCR. RESULTS: Amplified viral DNA was detected in each of the seven HIV-1-positive cases and in none of the five negative controls. In people with AIDS dementia, many HIV-1 DNA-positive cells were detected in regions of the CNS that corresponded to clinical symptomatology. In AIDS patients with minimal CNS involvement, rare HIV-1-infected microglial cells were noted. Viral RNA was detected primarily in cases of AIDS dementia. TNF, iNOS, MIP-1 alpha and MIP-1 beta expression localized to tissues from AIDS dementia cases where HIV-1 infected cells were plentiful. Colocalization experiments showed that these cytokines were transcribed mostly by viral-negative cells. CONCLUSIONS: These results suggest that two key elements in AIDS dementia are massive productive viral infection, involving microglia, neurons, and astrocytes, and concomitant stimulation of cytokine transcription in the neighboring uninfected cells.

Hereditary colobomatous anomalies of the optic nerve head.

The authors describe a family showing coloboma of the optic nerve associated with chorioretinal coloboma and coloboma of the iris. The absence of its occurrence in association with extraocular malformations points to an autosomal dominant mode of transmission, with reduced penetration. The authors describe the probable pathogenetic mechanism of the disorder and discuss the differential diagnosis both for chorioretinal coloboma (inflammatory chorioretinitis, especially by toxoplasma) and for colobomas of the optic disc (papillary pits and morning glory syndrome). The molecular basis of the malformation is however still unknown and thus a prenatal diagnosis is impossible.

Alcohol and poor compliance as factors in Wernicke's encephalopathy diagnosed 13 years after gastric bypass.

A severely obese man achieved rapid and significant weight loss after Roux-en-Y gastric bypass. Thirteen years later Wernicke's encephalopathy developed as a result of the patient's alcoholism, poor compliance with his micronutrient intake, poor oral intake and the decreased absorptive ability of the small bowel. In selecting any operation to treat morbid obesity the possibility of metabolic problems must be considered as well as the potential for substantial weight loss.

Congenital retinal macrovessel.

The authors describe a case of congenital retinal macrovessel with decreased visual acuity. Ophthalmoscopy and fluorescein angiography showed the malformation to be congenital and benign.