Identification and Construction of Strong Promoters in Yarrowia lipolytica Suitable for Glycerol-Based Bioprocesses.

Yarrowia lipolytica is a non-pathogenic aerobic yeast with numerous industrial biotechnology applications. The organism grows in a wide variety of media, industrial byproducts, and wastes. A need exists for molecular tools to improve heterologous protein expression and pathway reconstitution. In an effort to identify strong native promoters in glycerol-based media, six highly expressed genes were mined from public data, analyzed, and validated. The promoters from the three most highly expressed (H3, ACBP, and TMAL) were cloned upstream of the reporter mCherry in episomal and integrative vectors. Fluorescence was quantified by flow cytometry and promoter strength was benchmarked with known strong promoters (pFBA1in, pEXP1, and pTEF1in) in cells growing in glucose, glycerol, and synthetic glycerol media. The results show that pH3 > pTMAL > pACBP are very strong promoters, with pH3 exceeding all other tested promoters. Hybrid promoters were also constructed, linking the Upstream Activating Sequence 1B (UAS1B8) with H3(260) or TMAL(250) minimal promoters, and compared to the UAS1B8-TEF1(136) promoter. The new hybrid promoters exhibited far superior strength. The novel promoters were utilized to overexpress the lipase LIP2, achieving very high secretion levels. In conclusion, our research identified and characterized several strong Y. lipolytica promoters that expand the capacity to engineer Yarrowia strains and valorize industrial byproducts.

Chios mastic gum decreases renin levels and ameliorates vascular remodeling in renovascular hypertensive rats.

Chios mastic gum (CMG) exerts robust anti-inflammatory and antioxidant properties and it affects pathways that are implicated in the pathophysiology of endothelial and vascular inflammation. Aim of this study was to test the hypothesis that CMG administration lowers blood pressure (BP) and improves hypertension-induced target organ damage. 2-kidney, 1-clip (2K1C) hypertensive rats were treated with CMG (40 mg/kg body weight/day) for 2-weeks after the establishment of hypertension. Acute CMG administration lowered systolic, diastolic and mean arterial BP, while these hemodynamic effects were sustained throughout the 2-week administration period. CMG group also exhibited alleviated target organ damage as proposed by amelioration of biomechanical properties of the aorta -including cross-sectional area (CSA), aortic wall stiffness and thickness-, reversal of myocardial small vessel hypertrophy and maintenance of serum albumin levels. The anti-hypertensive effects of CMG are likely to be mediated by the decrease in renin serum levels. Regression analysis indicated that the effect of CMG on organ damage was BP-lowering dependent and was not associated with direct effects of renin or with its anti-inflammatory properties. We suggest a BP lowering effect of CMG via down-regulation of renin excretion associated with attenuation of target organ damage and inflammatory status. These observations provide profound evidence for the beneficial role of CMG in hypertension, which could possibly translate to further clinical research.

Effects of Chios mastic gum on cholesterol and glucose levels of healthy volunteers: A prospective, randomized, placebo-controlled, pilot study (CHIOS-MASTIHA).

BACKGROUND: Chios mastic gum (CMG) possesses anti-oxidant, anti-inflammatory, anti-atheromatic, lipid- and glucose-lowering properties. We evaluated the effects of CMG on cholesterol and fasting plasma glucose (FPG) levels of healthy volunteers. DESIGN: A prospective, randomized, placebo-controlled, pilot study. METHODS: One hundred and seventy nine volunteers with total cholesterol levels >200 mg/dl were randomized to four groups. Finally, 156 volunteers completed the follow-up period and were analysed: (1) control group (C, n = 23), receiving placebo; (2) total mastic (TM, n = 72) receiving daily a total dose of 1 g of crude CMG (330 mg capsules, tid); (3) polymer-free mastic (PFM, n = 33), receiving daily a total dose of 1 g of polymer free mastic (330 mg caps, tid); and (4) powder mastic (PM, n = 28), receiving daily a total dose of 2 g of crude CMG. RESULTS: After eight weeks, the TM group reduced total cholesterol by 11.5 mg/dl (p < 0.05) and FPG by 4.5 mg/dl (p < 0.05) adjusted for age, gender, BMI and baseline characteristics. The effect was stronger in overweight and obese patients (BMI > 25), with an estimated mean reduction of total cholesterol by 13.5 mg/dl (p < 0.05) and FPG by 5.1 mg/dl (p < 0.05). Administration of PFM and PM resulted in no statistically significant alteration. No effect was observed on LDL, HDL, triglycerides, uric acid and CRP. No gastrointestinal, liver or renal adverse events were recorded. CONCLUSIONS: CMG has a significant lowering effect on total cholesterol and glucose levels of healthy volunteers, with excellent tolerance and no detectable side effects, especially in overweight and obese individuals.

Beneficial health effects of Chios Gum Mastic and peroxisome proliferator-activated receptors: indications of common mechanisms.

For thousands of years, Chios Gum Mastic (CGM), the resin produced by the trunk of Pistachia lentiscus var Chia, has been used for culinary and medicinal purposes and several therapeutic properties have been attributed to it. CGM has been used in traditional medicine of various nations in the eastern Mediterranean area. This survey was carried out to identify biological mechanisms that could explain traditional usage and recent pharmacological findings. We reviewed the related scientific literature available from the NCBI PUBMED database on CGM studies and on natural products showing peroxisome proliferator-activated receptor (PPAR) agonist effects. We investigated whether CGM qualifies as a PPAR modulator. A large number of studies demonstrate that CGM has antioxidant, anti-inflammatory, hypolipidemic, and anticancer properties. Recently, the first evidence of CGM antidiabetic effect became known. CGM chemical composition has been extensively analyzed and the presence of several compounds, especially triterpenoids is well documented. Some of them, oleanonic acid, oleanolic acid, and gallic acid are considered to act as PPAR modulators. PPARs are nuclear receptors functioning as transcription factors and thereby controlling cellular functions at the level of gene expression. PPARs are involved in the pathways of significant diseases, such as metabolic syndrome, diabetes mellitus, dyslipidemia, inflammation, atheromatosis, and neoplasias, constituting a key target for pharmacological interventions. This article proposes that the synergistic action of some constituents of CGM on PPARs and more precisely on both PPARs isotypes-α and -γ, may be one of the major biological mechanisms via which CGM exerts its multiple effects.

Evaluation of Chios mastic gum on lipid and glucose metabolism in diabetic mice.

Chios mastic gum (MG), a resin produced from Pistacia lentiscus var. Chia, is reported to possess beneficial cardiovascular and hepatoprotective properties. This study investigated the effect of crude Chios MG on metabolic parameters in diabetic mice. Streptozotocin-induced diabetic 12-week-old male C57bl/6 mice were assigned to three groups: NC (n=9) control; LdM (n=9) animals receiving low dose mastic for 8 weeks (20 mg/kg body weight [BW]); and HdM (n=9) animals receiving high dose mastic (500 mg/kg BW) for the same period. Serum lipid and glucose levels were determined at baseline, at 4 and 8 weeks. Serum total protein, adiponectin, and resistin levels were also measured at the end of the experiment. Histopathological examination for liver, kidney, aorta, and heart lesions was performed. After 4 weeks, MG administration resulted in decreased serum glucose and triglyceride levels in both LdM and HdM, whereas BW levels were reduced in LdM group compared with controls. At the end of the experiment, LdM presented significantly lower serum glucose, cholesterol, low-density lipoprotein cholesterol, and triglyceride levels and improved high-density lipoprotein cholesterol levels compared with control group. HdM group had ameliorated serum triglyceride levels. Hepatic steatosis observed in control group was partially reversed in LdM and HdM groups. MG administered in low dosages improves glucose and lipid disturbances in diabetic mice while alleviating hepatic damage.

Forest fires are associated with elevated mortality in a dense urban setting.

OBJECTIVES: The climate and vegetation of the greater Athens area (population over three million) make forest fires a real threat to the environment during the summer. A few studies have reported the adverse health effects of forest fires, mainly using morbidity outcomes. The authors investigated the short-term effects of forest fires on non-accidental mortality in the population of Athens, Greece, during 1998-2004. METHODS: The authors used generalised additive models to investigate the effect of forest fires on daily mortality, adjusting for time trend and meteorological variables, taking into account air pollution as measured from fixed monitors. Forest fires were classified by size according to the area burnt. RESULTS: Small fires do not have an effect on mortality. Medium sized fires are associated with an increase of 4.9% (95% CI 0.3% to 9.6%) in the daily total number of deaths, 6.0% (95% CI -0.3% to 12.6%) in the number of cardiovascular deaths and 16.2% (95% CI 1.3% to 33.4%) in the number of respiratory deaths. Cardiovascular effects are larger in those aged <75 years, while respiratory effects are larger in older people. The corresponding effects of the one large fire are: 49.7% (95% CI 37.2% to 63.4%), 60.6% (95% CI 43.1% to 80.3%) and 92.0% (95% CI 47.5% to 150.0%). These effects cannot be completely explained by an increase in ambient particle concentrations. CONCLUSION: Forest fires have an immediate effect on mortality, not associated with accidental deaths, which is a significant public health problem, especially if the fire occurs near a densely populated area.

Aspergillus fumigatus abscesses of the thyroid with obstruction of the esophagus.

Thyroid invasion by Aspergillus spp. can occur with invasive aspergillosis, although it is rarely diagnosed antemortem. We describe a case of multiple thyroid abscesses from A. fumigatus that caused esophageal obstruction in a patient with myelodysplasia. Despite aggressive antifungal treatment, the outcome was rapidly fatal.