Asymmetric adaptation to indolic and aliphatic glucosinolates in the B and Q sibling species of Bemisia tabaci (Hemiptera: Aleyrodidae).

The role glucosinolates play in defending plants against phloem feeders such as aphids and whiteflies is currently not clear as these herbivores may avoid bringing glucosinolates from the phloem sap into contact with myrosinase enzymes. Here, we investigated the effects of high levels of aliphatic and indolic glucosinolates on life history traits and detoxification gene expression in two sibling species, B and Q, of the whitefly Bemisia tabaci. High levels of aliphatic glucosinolates decreased the average oviposition rate of both species and reduced the survival and developmental rate of Q nymphs. High levels of indolic glucosinolates decreased the oviposition rate and survival of nymphal stages of the B species and the developmental rate of both species. Molecular analyses revealed two major asymmetries between the B and Q species. First, specific GST genes (BtGST1 and BtGST2) were significantly induced during exposure to indolic glucosinolates only in Q. This may reflect the genes putative involvement in indolic glucosinolates detoxification and explain the species' good performance on plants accumulating indolic glucosinolates. Second, the constitutive expression of eight of the 10 detoxification genes analysed was higher in the Q species than in the B species. Interestingly, four of these genes were induced in B in response to high levels of glucosinolates. It seems, therefore, that the B and Q species differ in their 'optimal defence strategy'. B utilizes inducible defences that are profitable if the probability of experiencing the stress is small and its severity is low, while Q invests significant resources in being always 'ready' for a challenge.

Co-mapping studies of QTLs for fruit acidity and candidate genes of organic acid metabolism and proton transport in sweet melon (Cucumis melo L.).

Sweet melon cultivars contain a low level of organic acids and, therefore, the quality and flavor of sweet melon fruit is determined almost exclusively by fruit sugar content. However, genetic variability for fruit acid levels in the Cucumis melo species exists and sour fruit accessions are characterized by acidic fruit pH of <5, compared to the sweet cultivars that are generally characterized by mature fruit pH values of >6. In this paper, we report results from a mapping population based on recombinant inbred lines (RILs) derived from the cross between the non-sour 'Dulce' variety and the sour PI 414323 accession. Results show that a single major QTL for pH co-localizes with major QTLs for the two predominant organic acids in melon fruit, citric and malic, together with an additional metabolite which we identified as uridine. While the acidic recombinants were characterized by higher citric and malic acid levels, the non-acidic recombinants had a higher uridine content than did the acidic recombinants. Additional minor QTLs for pH, citric acid and malic acid were also identified and for these the increased acidity was unexpectedly contributed by the non-sour parent. To test for co-localization of these QTLs with genes encoding organic acid metabolism and transport, we mapped the genes encoding structural enzymes and proteins involved in organic acid metabolism, transport and vacuolar H+ pumps. None of these genes co-localized with the major pH QTL, indicating that the gene determining melon fruit pH is not one of the candidate genes encoding this primary metabolic pathway. Linked markers were tested in two additional inter-varietal populations and shown to be linked to the pH trait. The presence of the same QTL in such diverse segregating populations suggests that the trait is determined throughout the species by variability in the same gene and is indicative of a major role of the evolution of this gene in determining the important domestication trait of fruit acidity within the species.

Prevalence and fate of Giardia cysts in wastewater treatment plants.

The present study was conducted to review factors affecting the prevalence and concentration of Giardia in raw wastewater. The removal and inactivation efficiency of Giardia by wastewater treatment technologies was also reviewed. Data published for the prevalence of Giardia in wastewater and the removal by wastewater treatment plants was reviewed. Giardia cysts are highly prevalent in wastewater in various parts of the world, which may reflect the infection rate in the population. In 23 of 30 (76.6%) studies, all of the tested raw wastewater samples were positive for Giardia cysts at concentrations ranging from 0.23 to 100 000 cysts l(-1). The concentration of Giardia in raw wastewater was not affected by the geographical region or the socio-economic status of the community. Discharge of raw wastewater or the application of raw wastewater for irrigation may result in Giardia transmission. Activated sludge treatment resulted in a one to two orders of magnitude reduction in Giardia, whereas a stabilization pond with a high retention time removed up to 100% of the cysts from wastewater. High-rate sand filtration, ultrafiltration and UV disinfection were reported as the most efficient wastewater treatment methods for removal and disinfection of Giardia cysts. Wastewater treatment may not totally prevent the environmental transmission of Giardia cysts. The reviewed data show that a combination of wastewater treatment methods may results in efficient removal of Giardia cysts and prevent their environmental transmission.

Comparison of two treatments for the removal of selected organic micropollutants and bulk organic matter: conventional activated sludge followed by ultrafiltration versus membrane bioreactor.

The potential of membrane bioreactor (MBR) systems to remove organic micropollutants was investigated at different scales, operational conditions, and locations. The effluent quality of the MBR system was compared with that of a plant combining conventional activated sludge (CAS) followed by ultrafiltration (UF). The MBR and CAS-UF systems were operated and tested in parallel. An MBR pilot plant in Israel was operated for over a year at a mixed liquor suspended solids (MLSS) range of 2.8-10.6 g/L. The MBR achieved removal rates comparable to those of a CAS-UF plant at the Tel-Aviv wastewater treatment plant (WWTP) for macrolide antibiotics such as roxythromycin, clarithromycin, and erythromycin and slightly higher removal rates than the CAS-UF for sulfonamides. A laboratory scale MBR unit in Berlin - at an MLSS of 6-9 g/L - showed better removal rates for macrolide antibiotics, trimethoprim, and 5-tolyltriazole compared to the CAS process of the Ruhleben sewage treatment plant (STP) in Berlin when both were fed with identical quality raw wastewater. The Berlin CAS exhibited significantly better benzotriazole removal and slightly better sulfamethoxazole and 4-tolyltriazole removal than its MBR counterpart. Pilot MBR tests (MLSS of 12 g/L) in Aachen, Germany, showed that operating flux significantly affected the resulting membrane fouling rate, but the removal rates of dissolved organic matter and of bisphenol A were not affected.

Complication of laparoscopic detorsion of adnexal mass.

Detorsion of an ischemic adnexal mass has recently been advocated for most cases of twisted adnexa. Usually, the affected ovary regains some or all of its vitality and function. However, when the ovary is completely necrotic, it may form an abscess if it contains tissue components that cannot be eliminated by the peritoneal immune system. We report a case of pelvic abscess formation in a detorsed ovary that previously contained an unsuspected dermoid cyst. We call for an extensive inspection of the detorsed ovary before ending the laparoscopic operation, and if it remains necrotic and is suspected of containing a dermoid cyst, it should be removed promptly.

UV disinfection of wastewater effluents for unrestricted irrigation.

Wastewater reuse in arid regions is important for the production of a water resource to be utilised for non-potable purposes and to prevent the environmental transmission of disease-causing agents. This study was conducted to evaluate the effect of water quality on the comparative disinfection efficiency of viruses, bacteria and spores by UV irradiation. Furthermore, the microbial quality of effluent produced by coagulation, high rate filtration (HRF) and either UV irradiation or chlorination was determined. Using low pressure collimated beam, a UV dose of 80 mWs/cm2 was needed to achieve a 3-log10 inactivation of either rotavirus SA-11 or coliphage MS2, whereas over 5-log10 inactivation of E. coli was reached with a dose of only 20 mWs/cm2. B. subtilis inactivation was found to be linear up to a dose of 40 mWs/cm2 and then a tailing up to a UV dose of 120 mWs/cm2 was observed. It is worth noting that effluent turbidity of < 5 NTU did not influence the inactivation efficiency of UV irradiation. Operation of a pilot plant to treat secondary effluent by coagulation, HRF and UV disinfection at a UV dose of 80 mWs/cm2 resulted in the production of high quality effluent in compliance with the Israel standards for unrestricted irrigation (< 10 CFU/100 mL faecal coliform and turbidity of < 5 NTU). Sulphite reducing clostridia (SRC) were found to be more resistant than coliphages and F coliform for UV irradiation. The results of this study indicated that UV disinfection is suitable for the production of effluents for unrestricted irrigation of food crops.

Inter-ring communication is disrupted in the GroEL mutant Arg13 --> Gly; Ala126 --> Val with known crystal structure.

The crystal structures of the chaperonin GroEL Arg13 --> Gly; Ala126 --> Val double mutant, without and in complex with ATP gamma S, have been determined at atomic resolution. Here, we show that the double mutation Arg13 --> Gly; Ala126 --> Val disrupts negative co-operativity between GroEL rings, with respect to ATP, but has little effect on the positive co-operativity within each ring. Our results help to explain why the double mutation facilitated the crystallization of GroEL and why breaking of dyad symmetry between rings is not observed in crystal structures of this mutant. Our results may also help to explain why the observed structural differences between the GroEL double mutant and its ATP gamma S-bound form are small.

Manipulating duckweed through genome duplication.

Significant inter- and intraspecific genetic variation exists in duckweed, thus the potential for genome plasticity and manipulation is high. Polyploidy is recognised as a major mechanism of adaptation and speciation in plants. We produced several genome-duplicated lines of Landoltia punctata (Spirodela oligorrhiza) from both whole plants and regenerating explants using a colchicine-based cocktail. These lines stably maintained an enlarged frond and root morphology. DNA ploidy levels determined by florescence-activated cell sorting indicated genome duplication. Line A4 was analysed after 75 biomass doublings. Frond area, fresh and dry weights, rhizoid number and length were significantly increased versus wild type, while the growth rate was unchanged. This resulted in accumulation of biomass 17-20% faster in the A4 plants. We sought to determine if specific differences in gene products are found in the genome duplicated lines. Non-targeted ultra performance LC-quadrupole time of flight mass spectrometry was employed to compare some of the lines and the wild type to seek identification of up-regulated metabolites. We putatively identified differential metabolites in Line A65 as caffeoyl hexoses. The combination of directed genome duplication and metabolic profiling might offer a path for producing stable gene expression, leading to altered production of secondary metabolites.

The bitter side of the nightshades: Genomics drives discovery in Solanaceae steroidal alkaloid metabolism.

Steroidal alkaloids (SAs) and their glycosylated forms (SGAs) are toxic compounds largely produced by members of the Solanaceae and Liliaceae plant families. This class of specialized metabolites serves as a chemical barrier against a broad range of pest and pathogens. In humans and animals, SAs are considered anti-nutritional factors because they affect the digestion and absorption of nutrients from food and might even cause poisoning. In spite of the first report on SAs nearly 200 years ago, much of the molecular basis of their biosynthesis and regulation remains unknown. Aspects concerning chemical structures and biological activities of SAs have been reviewed extensively elsewhere; therefore, in this review the latest insights to the elucidation of the SAs biosynthetic pathway are highlighted. Recently, co-expression analysis combined with metabolic profiling revealed metabolic gene clusters in tomato and potato that contain core genes required for production of the prominent SGAs in these two species. Elaborating the knowledge regarding the SAs biosynthetic pathway, the subcellular transport of these molecules, as well as the identification of regulatory and signaling factors associated with SA metabolism will likely advance understanding of chemical defense mechanisms in Solanaceae and Liliaceae plants. It will also provide the means to develop, through classical breeding or genetic engineering, crops with modified levels of anti-nutritional SAs.

Hair chromium content of women with gestational diabetes compared with nondiabetic pregnant women.

Hair chromium concentration (HCC) of normal and diabetic pregnant women was determined by atomic-absorption spectroscopy. For nondiabetic pregnant women the value from 68 hair samples was 472 +/- 61 ng/g (mean +/- 95% CI); for gestational diabetics it was 734 +/- 155 ng/g from 42 hair samples. The difference was highly significant (P less than 0.005). Intermediate hair chromium concentrations were observed in 20 pregnant women with pregestational, overt diabetes mellitus (mean: 575 +/- 182 ng/g). Fifty-two women had a second hair sample taken later during pregnancy that showed a significant decrease in HCC (P less than 0.05). However, this decrease was confirmed only for the diabetic pregnant group. Age and parity did not influence the HCC. The data suggest that impaired utilization of chromium may be a possible etiology for gestational diabetes mellitus.

Prolactin response to electroconvulsive shock in anesthetized rats.

Serum prolactin level (SPL) of anesthetized rats was measured in response to electroconvulsive shock (ECS). This measurement served as an indicator of dopaminergic and serotonergic activity. Such neurotransmitter activity was modulated by pretreatment with cyproheptadine, parachlorophenylalanine (PCPA), pimozide, bromocriptine or L-dopa. A statistically significant lowered SPL was found in rats given ECS plus antiserotonergic agents, in comparison with animals receiving antiserotonergic agent or ECS alone. When ECS was administered to rats pretreated with the dopaminergic blocker, pimozide, there was a significant decrease in SPL, compared with animals receiving pimozide alone. When the dopaminergic system was stimulated by dopamine agonists, there was no elevation of SPL in response to ECS. Thus, there may be a balance between serotonergic and dopaminergic systems. The agents used in the experiment may upset this balance and produce a change in prolactin response to a given ECT stimulus.

The application of DNA microarrays in gene expression analysis.

DNA microarray technology is a new and powerful technology that will substantially increase the speed of molecular biological research. This paper gives a survey of DNA microarray technology and its use in gene expression studies. The technical aspects and their potential improvements are discussed. These comprise array manufacturing and design, array hybridisation, scanning, and data handling. Furthermore, it is discussed how DNA microarrays can be applied in the working fields of: safety, functionality and health of food and gene discovery and pathway engineering in plants.

Cytosolic aldolase is a ripening related enzyme in strawberry fruits (Fragaria x ananassa).

Two aldolase isoenzymes have been isolated from ripe strawberry fruits (Fragaria x ananassa cv. Camarosa and Elsanta) and partially purified by DEAE anion exchange and Sephacryl size exclusion chromatography. The isoenzymes were identified as class I cytosol and plastid aldolase on the basis of their chromatographic behavior on DEAE-cellulose columns, native molecular weight, pH optimum pattern, Km value for D-fructose-1,6-bisphosphate, tendency to be inactivated by lower pH values and SDS-PAGE subunit determination of 40 and 38 kDa, respectively. Total aldolase activity and distribution of both aldolase isoenzymes was also investigated at different stages of strawberry fruit ripening. Strawberries in the green and white ripening stage showed the same ratio of the two isoenzymes as green leaves with 15 and 8% cytosol aldolase activity, respectively. During strawberry fruit development the overall total aldolase activity decreased until the pink ripening stage and then increased due to a rise of cytosol aldolase yielding up to 75% in red strawberries. A cDNA putatively encoding the cytosolic form of aldolase in strawberry was cloned during the course of this study. Both microarray and RNA gel blot analyses showed that the cytosolic aldolase gene expression is induced during ripening as detected for the cytosolic aldolase enzyme. We suggest that induction of the cytosolic aldolase both at the levels of transcription and translation might be part of a ripening related stress response in the receptacle tissue.

Kallmann syndrome: a case of twin pregnancy and review of the literature.

In order to evaluate the possibilities for induction of ovulation, the functional competence of the pituitary gland of a woman with Kallmann syndrome was examined by two consecutive dynamic GnRH tests. The second test was conducted after 1 week's treatment by a GnRH pump. The results, which showed some rise of LH but no response of FSH, favored induction by hMG/hCG therapy. Three treatment cycles resulted in a twin pregnancy which was normal and was carried to term. Review of the literature shows only six previously reported pregnancies in women with Kallmann syndrome. Five of them were treated by hMG/hCG, and one by pulsatile GnRH. The two methods of induction are discussed in relation to the heterogeneity of the pituitary and ovarian function in Kallmann syndrome. We show that this heterogeneity dictates that the treatment for induction of ovulation should be individually adjusted according to the pituitary and ovarian competence.

Pulmonary complications in burn patients resuscitated with a low-volume colloid solution.

The occurrence of the adult respiratory distress syndrome (ARDS) and pneumonia was assessed in 482 patients with severe burns, but without evidence of inhalation injury. The patients were resuscitated during the burn shock period with a low-volume formula consisting mainly of plasma. The incidence of ARDS was 2.5 per cent for the entire burn population, and 9.4 per cent for 65 patients with burns covering more than 50 per cent of the body surface area. The general incidence of pneumonia was 4.4 per cent but was 12.5 per cent for the 65 patients with the extensive burns. These incidences were compared with other studies, and a pathophysiological explanation has been offered to explain differences in the frequency of pulmonary complications. The present review indicates that resuscitation with a low-volume formula consisting mainly of colloids can act as prophylaxis for reducing the incidence of ARDS and pneumonia.

Burn resuscitation with a low-volume plasma regimen--analysis of mortality.

The results of a mortality analysis from the Burn Center in Rambam Medical Center, Haifa, Israel, over a 5-year period are presented. The management of severe burns during the 'shock' period included a low-volume resuscitation formula consisting mainly of plasma and a small volume of crystalloid solution. The efficacy of burn management was assessed by probit analysis of the mortality data. We derived the percentage of body surface area burned associated with a 50 per cent probability of death (LA50) for four different age groups. The LA50 values obtained were compared to those obtained from other series; they compared favourably, demonstrating that resuscitation with low volumes consisting mainly of colloids may reduce complications associated with fluid overloading and therefore improve the LA50.

Early spontaneous rupture of the post myomectomy gravid uterus.

Rupture of a pregnant uterus is a serious threat to the mother's life and her fetus. Most of these cases have predisposing factors of which a post myomectomy scar is rare. Rupture of a post myomectomy gravid uterus usually occurs in the third trimester of pregnancy or during labor. We present a case of a very early spontaneous rupture which occurred at the 20th week of gestation in a post myomectomy uterus. To the best of our knowledge no previous report of a ruptured myomectomy scarred uterus has been described at such an early stage.

Prospective randomized comparison between an open and closed vaginal cuff in abdominal hysterectomy.

OBJECTIVE: To compare an operative and postoperative course of open vaginal cuff hysterectomy and closed vaginal cuff hysterectomy, and to correlate the length of stay, febrile morbidity and the incidence of pelvic fluid collections to the type of surgery. PARTICIPANTS: One-hundred women scheduled for hysterectomy were prospectively randomized into two groups that underwent either a closed or an open vaginal cuff technique. RESULTS: The open vaginal cuff technique took on average 19% more time than the closed vaginal cuff operation (P < 0.05, t-test). The incidence and size of pelvic fluid collections was significantly higher after the closed vaginal cuff hysterectomy than after the open technique (P < 0.01, t-test). However, the postoperative length of stay, febrile morbidity and the rate of complications were similar. CONCLUSIONS: Both techniques of hysterectomy produced a similar postoperative course despite the fact that the closed vaginal cuff technique resulted in a higher incidence of pelvic fluid collections. Therefore considering a shorter operation time for the closed vaginal cuff hysterectomy, this technique seems slightly preferable.

Biosynthesis of antinutritional alkaloids in solanaceous crops is mediated by clustered genes.

Steroidal glycoalkaloids (SGAs) such as α-solanine found in solanaceous food plants--as, for example, potato--are antinutritional factors for humans. Comparative coexpression analysis between tomato and potato coupled with chemical profiling revealed an array of 10 genes that partake in SGA biosynthesis. We discovered that six of them exist as a cluster on chromosome 7, whereas an additional two are adjacent in a duplicated genomic region on chromosome 12. Following systematic functional analysis, we suggest a revised SGA biosynthetic pathway starting from cholesterol up to the tetrasaccharide moiety linked to the tomato SGA aglycone. Silencing GLYCOALKALOID METABOLISM 4 prevented accumulation of SGAs in potato tubers and tomato fruit. This may provide a means for removal of unsafe, antinutritional substances present in these widely used food crops.