Effect of hydrogen peroxide on the activity and structure of Escherichia coli chaperone GroEL

Chaperone GroEL was treated with different concentrations of hydrogen peroxide. The conformational states of GroEL were monitored by protein intrinsic fluorescence, 8-anilino-1-naphthalene sulfonate fluorescence, and far-UV CD measurements. The results show that GroEL has unusual ability to resist oxidative stress. GroEL kept its quaternary structure and activity even when treated with 10 mM hydrogen peroxide. Two fragments were formed when GroEL was treated with high concentrations of hydrogen peroxide (more than 20 mM). It is suggested that GroEL, as a molecular chaperone, is related to oxidative process in vivo.     

An efficient method for the rapid establishment of Epstein-Barr virus immortalization of human B lymphocytes

Abstract. Several methods have been developed for the immortalization of B lymphocytes by Epstein-Barr virus (EBV). We developed an efficient method which reduces the time from culture initiation to immortalization and cryopreservation. Two infections of EBV to lymphocytes, and the use of phorbol ester-induced EBV stock significantly improved immortalization efficiency and reduced the time between initiation and immortalization and cryopreservation. The resulting cell bank was used to produce DNA for genetic studies focusing on the genes involved in immune and autistic disorders.     

Lipopolysaccharide pretreatment of the udder protects against experimental Escherichia coli mastitis

Exposure to pathogen-associated molecular patterns such as LPS can cause an immune refractory state in mammals known as endotoxin tolerance (ET), resulting in a decreased inflammatory response after pathogen contact. This ET concept was used to reduce the severity of an experimentally-induced clinical mastitis. Cows were pretreated with 1?μg LPS per udder quarter and challenged 72?h (group L72EC) or 240?h (group L240EC) later with 500 CFU Escherichia coli. Pretreated animals showed no leukopenia after challenge, no (L72EC), or only slightly (L240EC), elevated body temperature and significantly reduced systemic and local clinical scores compared with cows that were not pretreated. Whereas an increase of milk somatic cell count after the E. coli challenge was abrogated in L72EC animals, it was significantly delayed in the L240EC group. In both pretreated groups the bacterial load in milk was markedly reduced. Based on the expression of inflammation-related genes in lobulo-alveolar mammary tissue, the tolerizing effect of LPS pretreatment is based on the inhibited up-regulation of inflammatory (TNF-α, IL-6, CXCL8, CCL20) and anti-inflammatory genes (IL-10, IRAK-M). These findings indicate that the concept of ET may be usefully applied as mastitis prophylaxis facilitating a rapid response to microbial infection and avoiding dysregulated inflammation.     

Comparison of sampling methods for semipelagic animals in two deep basins of Lake Saimaa

The efficiency of different methods for sampling semipelagic animals was compared in the deep basins of Lake Paasivesi and Lake Puruvesi of the complex Lake Saimaa, Finland, during 1990–92. Certain semipelagic animals, such as Mysis relicta, amphipods and larvae of Chaoborus fiavicans, are important bioindicators of water and benthic quality. However, owing to their high mobility and vertical migrations they are difficult to sample quantitatively with traditional benthic methods. The equipment used for sampling was comprised of:

1. Vertical nets (0.56 and 0.13 m²) placed on the bottom for 5 minutes or 12 hours and hauled up vertically.
2. Plexiglass traps placed on the bottom for 12–14 hours.
3. Mysis-trawl hauled horizontally c. 250–300 metres on the bottom.
4. Ekman-grab.

In 1992, parallel samples were taken with vertically hauled nets (12 h and plexiglass traps baited with fish or liver or provided with a chemical light. Large vertically hauled nets were the most efficient equipment for catching Gammaracanthus lacustris, M. relicta and larvae of Ch. flavicans whilst baited or lighted nets were less effective. Lighted plexiglass traps were most effective in catching Pallasea quadrispinosa and water mites, but baited and control traps sampled inefficiently. The trawl sampled M. relicta and P. quadrispinosa relatively well, but was less efficient in sampling other groups. The Ekman-grab was effective only for sampling Monoporeia affinis. The semipelagic community differed between the studied basins. The dominant semipelagic animals in the humic, deep L. Paasivesi were M. relicta, amphipods and larvae of Ch. flavicans whilst the number of water mites was low in the catches. In the shallower, clear-water L. Puruvesi the densities of crustaceans and Chaoborus-larvae were low whilst some water mites, such as Piona pusilla, occurred in high numbers. Nocturnal sampling was more effective, especially for M. relicta.     

Influence of temperature on the development, reproduction and longevity of Ceratothripoides claratris (Thysanoptera: Thripidae) on tomatoes

Ceratothripoides claratris (Shumsher) is a serious pest attacking tomatoes in Thailand. Temperature-dependent development of C. claratris was studied at seven constant temperatures, i.e. 22, 25, 27, 30, 34, 35 and 40 degrees C. Pre-adult survivorship was greatest (95%) at 25 and 30 degrees C and shortest at 22 degrees C. Egg-to-adult time decreased within the range of 20 to 30 degrees C and at 34 degrees C it started to increase. The lower thermal threshold for egg-to-adult development was estimated at 16 and 18 degrees C by linear regression and the modified Logan model, respectively. The optimum temperature for egg-to-adult development was estimated at 32-33 degrees C by the modified Logan model. The influence of temperature on reproduction and longevity of C. claratris was determined at 25, 30 and 35 and 40 degrees C. Both inseminated and virgin females failed to reproduce at 40 degrees C. Virgin females produced only male offspring, confirming arrhenotoky. The sex ratio of the offspring of fertilized females was strongly female-biased, except at 25 degrees C. Mean total fecundity per female and mean daily total fecundity per female were highest for both virgin and inseminated females at 30 degrees C. Female longevity was longest at 25 degrees C and shortest at 40 degrees C. Male longevity was longest at 30 degrees C and shortest at 40 degrees C. The net reproductive rate (R0) and intrinsic rate of natural increase (rm) was greatest at 30 degrees C while, mean generation time (G) and the doubling time (t) were highest at 25 degrees C. The finite rate of increase (lambda) was fairly constant (1.1-1.5 days) over the three temperatures tested. The pest potential of C. claratris for tropical Asia is discussed.     

Prevalence and Correlates of Food Insecurity among Palestinian Refugees in Lebanon: Data from a Household Survey

Lebanon hosts the highest per capita refugee concentration worldwide. The Palestinian presence in Lebanon dates from 1948 and they remain a marginalized population. No information on their food security status has been reported previously. A survey of a representative sample of Palestinian refugee households in Lebanon (n = 2501) was conducted using a stratified two stage cluster sampling approach. We measured food insecurity using a modified USDA household food security module, locally validated. We collected data on household demographic, socioeconomic, health, housing, coping strategies and household intake of food groups and analysed these by food security status. About 41% (CI: 39-43) of households reported being food insecure and 20% (CI: 18-22) severely food insecure. Poor households were more likely to be severely food insecure (OR 1.41 (1.06-1.86)) while higher education of the head of household was significantly associated with protection against severe food insecurity (OR 0.66 (0.52-0.84)). Additionally, higher food expenditure and possession of food-related assets were significantly associated with food security (OR 0.93 (0.89-0.97) and OR 0.74 (0.59-0.92), respectively). After adjusting for confounders, households where at least one member suffered from an acute illness remained significantly more likely to be severely food insecure (OR 1.31(1.02-1.66)), as were households whose proxy respondent reported poor mental health (OR 2.64 (2.07-3.38)) and poor self-reported health (OR 1.62 (1.22-2.13). Severely food insecure households were more likely to eat cheaper foods when compared to non-severely food insecure households (p<0.001) and were more likely to rely on gifts (p<0.001) or welfare (p<0.001). They were also more likely to have exhausted all coping strategies, indicating significantly more frequently that they could not do anything (p = 0.0102). Food insecurity is a significant problem among Palestinian refugees in Lebanon and is likely to be exacerbated at this time when the Syrian crisis amplifies the problem.     

Effect of Zn2+ during reactivation and refolding of urea-denatured creatine kinase

The courses of refolding and reactivation of urea-denatured creatine kinase (CK) (ATP:creatine N-phosphotrans-ferase, EC 2.7.3.2) have been studied in the absence and presence of zinc ions. The presence of Zn²⁺ at low concentrations blocks the reactivation and refolding of urea-denatured CK and keeps it in a partially folded state. The partially folded state proved to be a monomeric state which resembles the molten globule state in the CK folding pathway. During refolding in the presence of Zn²⁺ , creatine kinase forms aggregates with the aggregation dependent on zinc concentration and temperature. In the presence of EDTA, the partially folded creatine kinase can be reactivated and refolded following a biphasic course, suggesting the existence of a monomeric intermediate during the refolding of CK. The results also suggest that low concentrations of zinc ions might be toxic to some proteins such as creatine kinase by disrupting their proper folding.     

Liposomal cargo unloading induced by pH-sensitive peptides.

Amphiphilic peptides designed to have a pH-dependent conformational change and membrane activity are described. At physiologic pH, the peptides would exist in a random coil conformation, but at endosomal pH values they would switch to amphiphilic alpha-helices, disrupt membranes, and release liposomal contents. A series of peptides have been investigated that contain a high percentage of Glu residues for the pH-induced conformational switch, and Leu residues for optimal lipid binding. Circular dichroism (CD) results in aqueous and liposomal environments were performed and demonstrate a pH-dependent shift to helicity upon acidification. Liposomal release data at neutral and acidic pH, also document the success of this design strategy.