Leishmania species: mechanisms of complement activation by five strains of promastigotes

The interaction of fresh serum with promastigotes of Leishmania major, L. donovani, L. mexicana mexicana, L. mexicana amazonensis, and L. braziliensis guyanensis results in lysis of all strains tested with either fresh human or guinea pig serum at 37 C for 30 min. Lysis does not occur in the cold and requires divalent cations and complement that is active hemolytically. Serum deficient in the eighth component of complement is not lytic. Lysis of L. major, L. mexicana, and L. braziliensis proceeds fully in human serum containing EGTA/Mg2+ or in guinea pig serum deficient in the fourth complement component. These species consume only small amounts of C4 from human serum and do not require calcium to optimally bind C3. The data indicate that all are activators of the alternative complement pathway and that the classical pathway is not required for the lysis of these organisms. Promastigotes of L. donovani, in contrast, activate the classical pathway. The presence of calcium is required for both optimal C3 binding and parasite lysis, and L. donovani promastigotes consume C4 when incubated in human serum. In high concentrations, human serum agglutinates all tested Leishmania spp. The agglutinating factor does not require divalent cations, is heat stable, and works at 4 C, suggesting that it is an antibody. This "naturally occurring" antibody cross reacts with all Leishmania spp. and agglutinates them. The adsorption of serum with any Leishmania species or with beads that are Protein A coated, removes the agglutinogen. This factor causes a slight enhancement in alternative pathway activation by L. major and mediates the classical activation by L. donovani. In adsorbed serum, L. donovani promastigotes only weakly activate the alternative complement pathway. Increased concentrations of adsorbed serum are therefore necessary for lysis to proceed. The titer can be partially restored by the addition of heat inactivated serum. Using purified components of the classical cascade, we are unable to visualize surface bound C3 on L. donovani promastigotes unless heat inactivated serum is also present. We conclude that all Leishmania spp. promastigotes are susceptible to lysis by normal serum independent of antibody. The presence of small amounts of naturally occurring antibody in human serum enhances the susceptibility of L. donovani promastigotes to lysis by activating the classical complement pathway.     

Microbiological quality of tomato (<i>Solanum lycopersicum</i> L.) produced under greenhouse conditions in five Municipalities of the State of Mexico

The aim of the current research was to determine tomato (Solanum lycopersicum L.) microbiological quality produced under greenhouse conditions in 5 municipalities of the State of Mexico. Studies were conducted during the 2013 production cycle to know the risks and apply prevention strategies prior to its consumption. A microbiological analysis of samples of irrigation water, soil and 100 tomato fruits variety cid was performed to determine Aerobic Mesophiles, Total Coliforms and Fecal Coliforms. The methodology used were those according to the Official Mexican Standards NOM- 109-SSA1-1994, NOM-110-SSA1-1994, NOM-092-SSA1-1994, NOM-113-SSA1-1994, and the Regulations of the National French Organization for Standardization (AFNOR) NF V08-60, and NOM-093-SSA1-1994, which establish the allowable limits for the study microorganisms. The results showed a zero level of pollution in water and soil samples. For fruits, levels of Aerobic Mesophilic were within the maximum limits permitted by the standards. The municipality of Texcaltitlan showed the highest average for these microorganisms (10083.80 CFU/mL). Huixquilucan showed 2266.84 CFU/mL for Total Coliforms. For Fecal Coliforms, municipalities of Coatepec and Texcaltitlan exceeded the allowed limit.     

A significant range extension and sanctuary for the rare Open Bay Island skink (Oligosoma taumakae)

Abstract

The Open Bay Island skink (Oligosoma taumakae) is one of New Zealand's rarest lizard species. Until 2010, it was known only from two small islands in the Open Bay Island Group, a Māori-owned wildlife sanctuary in South Westland, New Zealand. Skinks on these islands are threatened by predation from weka (Gallirallus australis), a flightless native rail thought to have been introduced to the Open Bay Islands c. 100 years ago. Here, we describe the discovery of Open Bay Island skinks on two vegetated rock stacks located off the coast of Barn Bay, 52 km southwest of the Open Bay Islands. Although small (c. 0.10 and 0.36 ha), the Barn Islands appear to be predator-free, providing an important sanctuary for the skinks. We recommend: (1) a survey of mainland sites with suitable habitat; and (2) an evaluation of the need for island biosecurity measures for detecting and responding to incursions of small mammals.     

Effect of temperature on efficacy of insecticides to differential grasshopper (Orthoptera: Acrididae)

The effect of temperature on activity of insecticides for controlling grasshoppers in leafy green vegetables was evaluated. Insecticides evaluated had differing modes of action and included diflubenzuron, azadirachtin, Beauveria bassiana, spinosad, endosulfan, esfenvalerate, and naled. We evaluated these insecticides for efficacy to third instars of differential grasshopper, Melanoplus differentialis (Thomas), at temperatures ranging from 10 to 35 degrees C. In the laboratory, treatment with esfenvalerate resulted in 100% mortality at temperatures of 10 to 35 degrees C, and efficacy was not temperature dependent. Treatment with spinosad resulted in similar mortality as with esfenvalerate at all temperatures except 10 degrees C. The activity of B. bassiana was greatest at 25 degrees C and was adversely affected by high and low temperatures. Treatment with diflubenzuron resulted in increased mortality at high temperatures, and at 35 degrees C its activity was similar to that of esfenvalerate and spinosad. The activity of azadirachtin ranged from 19 to 31% and was not influenced by temperature. In field studies, spinosad, diflubenzuron, and esfenvalerate provided differing levels of mortality both at application and when nymphs were exposed to 1-h-old residues. However, only spinosad and diflubenzuron provided similar levels of mortality when nymphs were exposed to 24-h-old residues. The residual activity of endosulfan, naled, esfenvalerate, and spinosad decreased with increasing time (0-24 h) after exposure to sunlight and high summer temperatures. Compared with other insecticides, naled had a short residual activity period and activity was dependent upon immediate contact with the nymphs or their substrate. B. bassiana was inactive under high temperatures and intense sunlight as occurs in summer.     

Delivery of photoreactive psoralen derivatives to specific biological targets

Psoralens are bifunctional molecules which photoreact with the pyrimidine bases of nucleic acids to form monoadducts and diadducts, or interstrand cross-links. We have prepared psoralen derivatives with additional functional groups which can be specifically directed to chosen biological targets. A sulfhydryl-containing psoralen which can form site-specific cross-links in plasmid DNA has been used to study psoralen repair and mutagenesis. Cloned DNA containing psoralen monoadducts has been cross-linked to specific regions of viral RNA and used to probe virus assembly. A biotinylated psoralen derivative which binds specifically to avidin has been used to detect small amounts of DNA. Finally, a psoralen derivative of insulin has been used to deliver psoralen specifically to activated lymphocytes.     

The roles of degranulation and superoxide anion generation in neutrophil aggregation

Human neutrophils when exposed to appropriate stimuli aggregate, generate O(2) and secrete lysosomal constituents. To determine whether a causal relationship may exist between these responses neutrophils were exposed to either N-formyl-methionyl-leucyl-phenylalanine, phorbol myristate acetate, or the two calcium ionophores, A23187 and prostaglandin Bx. Each agent elicited all of the above responses. The concentrations required to elicit the aggregation of 30 . 10(6) neutrophils/ml were comparable to that required for O(2) generation or lysozyme release. In a series of experiments designed to dissociate these responses, cells were suspended in a concentration too dilute (3 . 10(6) neutrophils/ml) to permit aggregation to occur. O(2) generation and lysozyme release was measurable and varied in a dose-dependent fashion to the concentration of stimulus. In a second series of experiments, neutrophils were treated with 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid to inhibit degranulation without affecting O(2) generation. Aggregation was inhibited in a parallel fashion with lysozyme release. When detectable O(2) was removed from the medium by superoxide dismutase and catalase, aggregation and lysozyme release unaffected showing that aggregation can not be due to the presence of O(2) or its products in the extracellular medium. Neither aggregation of resting cells nor augmentation of fMet-Leu-Phe-induced aggregation was observed when cells were exposed to either supernatants of degranulated neutrophils or constituents of specific granules (lysozyme, lactoferrin). Kinetic analysis showed that in the absence of cytochalasin B degranulation preceded aggregation, while in its presence aggregation preceded degranulation.     

Bilateral detection thresholds in dextrals and sinistrals reflect the more sensitive side of the nose, which is not lateralized [published erratum appears in Chem Senses 1998 Dec;23(6):761]

Several fundamental questions remain enigmatic concerning human olfactory sensitivity, including (i) whether detection threshold differences exist between the two sides of the nose (and, if so, whether such differences are influenced by handedness) and (ii) whether bilateral (i.e. binasal) stimulation leads to lower thresholds than unilateral stimulation (and, if so, whether the degree of facilitation is inversely related to general olfactory ability). In this study, and well-validated single staircase procedure was used to establish bilateral and unilateral detection thresholds for the cranial nerve I stimulant phenyl ethyl alcohol in 130 right- and 33 left-handed subjects. No differences in sensitivity between the left and right sides of the nose were observed in either group. Bilateral thresholds were lower, on average, than unilateral thresholds when the latter were categorized in terms of left and right nares. However, the bilateral thresholds did not differ significantly from those of the side of the nose with the lower threshold. Overall smell ability, as measured by the University of Pennsylvania Smell Identification Test, did not interact with any of the test measures. These data imply that (i) the left and right sides of the nose do not systematically differ in detection threshold sensitivity for either dextrals or sinistrals and (ii) if central integration of left:right olfactory threshold sensitivity occurs, its effects do not exceed the function of the better side of the nose.      

Possible congenital hemolytic anemia in prehistoric coastal inhabitants of Israel.

The spread of thalassemia among prehistoric populations of the Mediterranean Basin has been linked to the increased risk to early agriculturalists posed by the Plasmodium falciparum parasite. The diagnosis of the disease in human skeletal remains, however, has usually been based on a single pathological criterion, porotic hyperostosis. This paper reports on what we believe to be the earliest case of thalassemia yet identified in the prehistoric record. Our diagnosis of the disease in an individual from the submerged Prepottery Neolithic B village of Atlit-Yam off the Israeli coast is based on a pathological humerus demonstrating a pattern of deformation characteristic of clinical thalassemia. The implications of these findings for our understanding of human societies undergoing the transition from foraging to agriculture in the Near East are discussed.