Biomonitoring of workers cleaning up ammunition waste sites

2,4,6-Trinitrotoluene (TNT) is an important occupational and environmental pollutant. In TNT-exposed humans, notable toxic manifestations have included aplastic anaemia, toxic hepatitis, cataracts, hepatomegaly and liver cancer. Therefore, it is important to develop protection measures and to monitor workers involved in the clean-up of ammunition sites. Haemoglobin (Hb) adducts of TNT, 4-amino-2,6-dinitrotoluene (4ADNT) and 2-amino-4,6-dinitrotoluene (2ADNT), and the urine metabolites of TNT, 4ADNT and 2ADNT were found in 22–50% of the exposed workers, but not in the control group. The exposed workers were wearing protective equipment. The levels of erythrocytes, haemoglobin, creatinine, serum glutamic pyruvic transaminase and lymphocyte levels were significantly lower in the exposed workers than in the non-exposed workers. The levels of blood urea and reticulocytes were significantly higher in the exposed workers than in the non-exposed workers. Headache (26%), mucous membrane irritation (16%), sick leave (18%), lassitude (8%), anxiety (6%), shortness of breath (3%), nausea (5%) and allergic reactions (8%) were reported by the exposed workers. In a further analysis the U-4ADNT levels and the Hb-adduct levels were compared to the blood parameter and the health effects. The blood parameters were not significantly different between the U-4ADNT positive and U-4ADNT-negative group. Headache, mucous membrane irritation, sick leave, lassitude, anxiety, shortness of breath and allergic reactions were statistically not different between the two groups. Also in the workers with Hb-4ADNT adducts no significant negative changes were seen in regards to the changes of the blood parameters or the health effects. According to the results of the present study, it appears that the blood parameter changes and the health effects are more influenced by other factors than by the internal exposure to TNT.     

Retinoic acid and/or progesterone differentiate mouse induced pluripotent stem cells into male germ cells in vitro

Numerous reagents were employed for differentiating induced pluripotent stem cells (iPSCs) into male germ cells; however, the induction procedure was ineffective. The aim of this study was to improve the in vitro differentiation of mice iPSCs (miPSCs) into male germ cells with retinoic acid (RA) and progesterone (P). miPSCs were differentiated to embryoid bodies (EBs) in suspension with RA with or without progesterone for 0, 4, and 7 days. Then, the expression of certain genes at different stages of male germ cell development including Ddx4 (pre meiosis), Stra8 (meiosis), AKAP3 (post meiosis), and Mvh protein was examined in RNA and/or protein levels by real-time polymerase chain reaction or flow cytometry, respectively. The Stra8 gene expression increased in the RA groups on all days. But, expression of this gene declined in RA + P groups. In addition, an increased expression of Ddx4 gene was observed on day 0 in the P group. Also, a significant upregulation was observed in the expression of AKAP3 gene in the RA + P group on days 0 and 4. However, gene expression decreased in P and RA groups on day 7. The expression of Mvh protein significantly increased in the RA group on day 7. The Mvh expression was also enhanced in the P group on day 4, but it decreased on day 7, while this protein upregulated on day 0 and 7 in the RA + P group. The miPSCs have the capacity for in vitro differentiation into male germ cells by RA and/or progesterone. However, the effects of these inducers depend on the type of combination and an effective time.     

Mass spectrometry imaging: An emerging technology for the analysis of metabolites in insects

Mass spectrometry imaging (MSI) can visualize the composition, abundance, and spatial distribution of molecules in tissues or cells, which has been widely used in the research of life science. Insects, especially the agricultural pests, have received a great deal of interests from the scientists in biodiversity and food security. This review introduces the major characteristics of MSI, summarizes its application to the investigation of insect endogenous metabolites, exogenous metabolites, and the spatiotemporal changes of metabolites between insects and plants, and discusses its shortfalls and perspectives. The significance of these concerns is beneficial for future insect research such as physiology and metabolism.     

Perylenequinones from Curvularia lunata

Three perylenequinones, the methylated 12-epi-stemphytriol (1), dihydroalterperylenol (2), and alterperylenol (3), were isolated from cultures of Curvularia lunata (LBQM-04) on malt extract broth. All these compounds were obtained from this particular fungus for the first time. Structures of compounds were determined based on MS and NMR spectra, as well as by comparison with the literature reports. The isolation of these phytotoxic reduced perylenequinones from the Curvularia genus suggested that they may occur in any anamorphic fungi belonging to the Pleosporaceae family.     

Influence of carbon and nitrogen sources on antifungal metabolite production by bacterium associated with entomopathogenic nematode against Penicillium expansum

A specific symbiotic Bacillus sp. isolated from a rhabditid entomopathogenic nematode, Rhabditis (Oscheius) sp. was found to produce large number of bioactive compounds. The present study was conducted to determine the effect of carbon and nitrogen sources for the production of antimicrobial substances by Bacillus sp. The yield of the crude antimicrobial substances and antimicrobial activity against the test micro-organism also differed significantly when carbon and nitrogen sources in the fermentation media were changed. The antifungal activity was significantly high in yeast extract plus fructose (46.5?±?2.12?mm) followed by yeast extract plus maltose, beef extract plus fructose and meat infusion plus glucose. High pressure liquid chromatography analysis of the crude antimicrobial substances revealed different peaks with different retention time indicating that they produced different compounds. When the carbon source was not included in the fermentation media, the antimicrobial production was substantially reduced. The results indicate that carbon source in the fermentation media plays a vital role in the production of antifungal substances. It is concluded that yeast extract and fructose as nitrogen and carbon sources produced maximum activity, which can effectively control the blue mould caused by Penicillium expansum in apples and pears.     

Antagonistic effect of two isolates of Trichoderma harzianum against postharvest pathogens of tomato (Lycopersicon esculentum)

Pathogenicity test of all fungi (14 different isolates) isolated from both infected tomato fruits and the surface wash of other healthy fruits had different pathogenicity rates. The genus Rhizopus sp. was the most pathogenic one followed by Fusarium sp. Trichoderma harzianum isolates (T3 and T4) had a different pattern of antagonism against the tested pathogens. In dual plate test of the antagonistic action of T3 and T24 against the postharvest pathogens, clear zone size ranged between 1 and 4 and 3 and 6 mm by T3 and T24, respectively. The antagonists (T3 and T24) didn't show inhibition zone against Rhizopus sp., but they could overgrow it by 100% after 9 days of incubation. Mostly, all the other postharvest isolates showed high degree of overgrowth by T3 than T24. The two antagonists failed to overgrow Aspergillus species except for A. niger (3) which was overgrown by T3. Volatile and non-volatile metabolite tests indicated that mycelial growth of Penicillium stekii was significantly inhibited by T3 and T24 more than the other tested pathogens. The inhibition of A. niger (1) was 12% by non-volatile metabolites of T24 produced after 1 day incubation, and reached to 97% inhibition by the metabolites of 3 days. Interestingly, inhibition of Aspergillus sp. by volatile compounds of T3 and T24 was 2% and 20%, respectively, whereas the inhibition of the same pathogen by non-volatile compounds reached 75% and 87%, respectively. The results of slice assay clearly indicate that T. harzianum (T3 and T24) could provide a complete protection to tomato slices from the infection of the tested pathogens. After 3 days of incubation, Trichoderma suppressed the linear growth of these pathogens on tomato slices and the percentage of suppression was significant and ranged between 80 and 100%, except with Rhizopus sp. the suppression reached 33% only.     

Solid-State Fermentation Systems—An Overview

Abstract

Starting with a brief history of solid-state fermentation (SSF), major aspects of SSF are reviewed, which include factors affecting SSF, biomass, fermentors, modeling, industrial microbial enzymes, organic acids, secondary metabolites, and bioremediation. Physico-chemical and environmental factors such as inoculum type, moisture and water activity, pH, temperature, substrate, particle size, aeration and agitation, nutritional factors, and oxygen and carbon dioxide affecting SSF are reviewed. The advantages of SSF over Submerged Fermentation (SmF) are indicated, and the different types of fermentors used in SSF described. The economic feasibilities of adopting SSF technology in the commercial production of industrial enzymes such as amylases, cellulases, xylanase, proteases, phytases, lipases, etc., organic acids such as citric acid and lactic acid, and secondary metabolites such as gibberellic acid, ergot alkaloids, and antibiotics such as penicillin, cyclosporin, cephamycin and tetracyclines are highlighted. The relevance of applying SSF technology in the production of mycotoxins, biofuels, and biocontrol agents is discussed, and the need for adopting SSF technology in bioremediation of toxic compounds, biological detoxication of agro-industrial residues, and biotransformation of agro-products and residues is emphasized.     

ALTERATIONS WITH AGING OF THE ENDOCRINE AND NEUROENDOCRINE CIRCADIAN SYSTEM IN HUMANS

This was an open-label study in 19 children aged 9–13 years, weighing 27–44 kg, with bronchial asthma. Twenty-four-hour steady-state concentrations of theophylline and its metabolites 1,3-dimethyl uric acid, 3-methyl xanthine and 1-methyl uric acid were assessed after daily dosing of 600 mg (ca18 mg/kg/day) of the sustainedrelease theophylline micro-pellet sprinkle system BY158K, for 4 days. The dosing regimen used was an unequal twice-daily dose of 200 mg in the morning after breakfast and 400 mg in the evening after dinner. Twenty-four-hour peak expiratory flow (PEF) profiles were compared before treatment and at steady-state, along with lung function parameters after bronchial provocation. Mean values±SD (n=16) of the steady-state characteristics were C_min6.8±2.1 mg/1, C_max14.5±4.8 mg/1 and C_av10.S±2.9 mg/1, the plateau time was 11.7±4.8 hr and peak-trough fluctuation and swing were 72±21 and 118±52%, respectively. There was an excellent reproducibility of theophylline pre-dose levels at corresponding time points of the 24-hr sampling period [r=0.864 (p< 0.001)]. Mean values±SD of the 24 hr average serum metabolite levels were 0.9±0.2 mg/1 for 1, 3-dimethyl uric acid, 0.6±0.1 mg/1 for 3-methyl xanthine and 0.4±0.1 mg/1 for 1-methyl uric acid. Lung function (n=17) following bronchial provocation, improved in 10 children after theophylline treatment of 4 days, remained stable in 2 patients and deteriorated in 5 patients. Serum theophylline profiles and PEF profiles ran largely in parallel over the 24-hr period. Six children exhibited typical theophylline induced side-effects, headache (n=3), nausea (n=4), dizziness (n=l), vomiting (n=4), sleep disturbances (n=1), pallor (n=1) and tremor(n=1), necessitating in 3 children one dose omission/reduction (n=2) or subsequent dose reduction (n=1). It has been shown that a twice daily dosing regimen with unequal doses of anhydrous theophylline (BY158K) is well suited to this population of fast metabolisers. The patients were well protected throughout the day, including the critical early morning hours.