Pollination and fruit set in pumpkin (Cucurbita pepo) by honey bees

Species of Cucurbitaceae are cultivated worldwide and are depend on bee pollination for fruit set. Field and lab experiments were conducted at Cornell University, Ithaca, NY, during 1996 and 1997 to determine "Howden" pumpkin (Cucurbita pepo L.) pollen removal and deposition by honeybees and factors relating to male flower attractiveness. Several parameters were evaluated in flowers at anthesis: (1) removal of pollen from anthers by honey bees, (2) pollen deposition on the stigma by honey bees, (3) amount of pollen on the body of honey bees, (4) fruit set after bee pollination, and (5) male flower nectary's pores and flower attractiveness. Honey bees carried between 1,050 to 3,990 pollen grains and 13,765 were removed from an anther after one visit. The amount of pollen deposited on the stigma by the honey bees varied according to the number of visits, from 53 grains with one visit, to 1,253 grains with 12 visits, and the mean number of grains in each visit varied from 53 to 230 grains. The percentage of established fruits was higher (100%) when the flowers received 12 visits of Apis mellifera, corresponding to a load 1,253 pollen grains. The attractiveness of the male flower for pollen and nectar collection was increased by the degree of opening of the access pore to the nectary in the flower.

Morphology and anatomy of the developing fruit of Maclura tinctoria, Moraceae

(Morphology and anatomy of the developing fruit of Maclura tinctoria, Moraceae). Maclura tinctoria (L.) D. Don ex Steudel was selected for the present study due of its economic and medicinal importance. The purpose of this investigation is to present a detailed description of the fruit development, specially by: (a) defining the fruit type presented by the species, and (b) characterizing the seed type of the species based upon the presence or not of mechanical tissue on the seed-coat. The fruit originates from the subglobose female inflorescence which consists of small unipistillate flowers with superior ovary, unilocular and uniovular apical placentation. The mature fruit is multiple, constituted of small drupes. The ovule is ana-campylotropous, suspended, bitegmic and crassinucellate. The mature seed is flattened, slightly ovated, cream colored, with unspecialized membrane coat with thin-walled cells more or less crushed. The seed has parenchymatic endosperm with lipophilic content. The embryo is straight, with two cotyledons of the same size. Ontogenetic studies reveal that the fruits are infrutescences. The fleshy edible part is derived from the perigone and inflorescence axis. The drupes consist of a single pyrene of macrosclereids.

Effect of bacterial lipopolysaccharide on gastric emptying of liquids in rats

The objectives of the present investigation were 1) to study the effect of bacterial lipopolysaccharide (LPS) on rat gastric emptying (GE) and 2) to investigate a possible involvement of the vagus nerve in the gastric action of LPS. Endotoxin from E. coli (strain 055:B5) was administered sc, ip or iv to male Wistar rats (220-280 g body weight) at a maximum dose of 50 µg/kg animal weight. Control animals received an equivalent volume of sterile saline solution. At a given time period after LPS administration, GE was evaluated by measuring gastric retention 10 min after the orogastric infusion of a test meal (2 ml/100 g animal weight), which consisted of 0.9% NaCl plus the marker phenol red (6 mg/dl). One group of animals was subjected to bilateral subdiaphragmatic vagotomy or sham operation 15 days before the test. A significant delay in GE of the test meal was observed 5 h after iv administration of the endotoxin at the dose of 50 µg/kg animal weight. The LPS-induced delay of GE was detected as early as 30 min and up to 8 h after endotoxin administration. The use of different doses of LPS ranging from 5 to 50 µg/kg animal weight showed that the alteration of GE was dose dependent. In addition, vagotomized animals receiving LPS displayed a GE that was not significantly different from that of the sham control group. However, a participation of the vagus nerve in LPS-induced delay in GE could not be clearly demonstrated by these experiments since vagotomy itself induced changes in this gastric parameter. The present study provides a suitable model for identifying the mechanisms underlying the effects of LPS on gastric emptying

Fibronectin in the ascitic fluid of cirrhotic patients: correlation with biochemical risk factors for the development of spontaneous bacterial peritonitis

Cirrhotic patients (23 with alcoholic cirrhosis, 5 with posthepatitic cirrhosis and 2 with cryptogenic cirrhosis) with ascites and portal hypertension were studied and divided into two groups corresponding to high or low risk to develop spontaneous bacterial peritonitis (SBP) related to the concentration of total protein in the ascitic fluid (A-TP): group I (high risk): A-TP£1.5 g/dl and group II (low risk): A-TP>1.5 g/dl. Fibronectin (FN), C3 and C4 concentrations were measured by radial immunodiffusion while total protein was measured by the biuret method. The mean values (group I vs group II) of C3 (12.59 ± 4.72 vs 24.53 ± 15.58 mg/dl), C4 (4.26 ± 3.87 vs 7.26 ± 4.14 mg/dl) and FN (50.47 ± 12.49 vs 75.89 ± 24.70 mg/dl) in the ascitic fluid were significantly lower (P<0.05) in the group considered to be at high risk for SBP. No significant difference was observed in the plasma/ascites fibronectin ratio (3.91 ± 1.21 vs 3.80 ± 1.26) or gradient (131.46 ± 64.01 vs 196.96 ± 57.38) between groups. Fibronectin in ascites was significantly correlated to C3 (r = 0.76), C4 (r = 0.58), total protein (r = 0.73) and plasma FN (r = 0.58) (P<0.05). The data suggest that the FN concentration in ascites is related to the opsonic capacity of this fluid, and that its concentration in the ascitic fluid may be a biochemical risk factor indicator for the development of spontaneous bacterial peritonitis

The role of T cell subsets and cytokines in the regulation of intracellular bacterial infection

Cellular immune responses are a critical part of the host's defense against intracellular bacterial infections. Immunity to Brucella abortus crucially depends on antigen-specific T cell-mediated activation of macrophages, which are the major effectors of cell-mediated killing of this organism. T lymphocytes that proliferate in response to B. abortus were characterized for phenotype and cytokine activity. Human, murine, and bovine T lymphocytes exhibited a type 1 cytokine profile, suggesting an analogous immune response in these different hosts. In vivo protection afforded by a particular cell type is dependent on the antigen presented and the mechanism of antigen presentation. Studies using MHC class I and class II knockout mice infected with B. abortus have demonstrated that protective immunity to brucellosis is especially dependent on CD8+ T cells. To target MHC class I presentation we transfected ex vivo a murine macrophage cell line with B. abortus genes and adoptively transferred them to BALB/c mice. These transgenic macrophage clones induced partial protection in mice against experimental brucellosis. Knowing the cells required for protection, vaccines can be designed to activate the protective T cell subset. Lastly, as a new strategy for priming a specific class I-restricted T cell response in vivo, we used genetic immunization by particle bombardment-mediated gene transfer

The effect of bacterial lipopolysaccharide on gastric emptying in rats suffering from moderate renal insufficiency

The objective of the present study was to evaluate the response of rats suffering from moderate renal insufficiency to bacterial lipopolysaccharide (LPS, or endotoxin). The study involved 48 eight-week-old male SPF Wistar rats (175-220 g) divided into two groups of 24 animals each. One group underwent 5/6 nephrectomy while the other was sham-operated. Two weeks after surgery, the animals were further divided into two subgroups of 12 animals each and were fasted for 20 h but with access to water ad libitum. One nephrectomized and one sham-treated subgroup received E. coli LPS (25 µg/kg, iv) while the other received a sterile, pyrogen-free saline solution. Gastric retention (GR) was determined 10 min after the orogastric infusion of a standard saline test meal labeled with phenol red (6 mg/dl). The gastric emptying of the saline test meal was studied after 2 h. Renal function was evaluated by measuring the plasma levels of urea and creatinine. The levels of urea and creatinine in 5/6 nephrectomized animals were two-fold higher than those observed in the sham-operated rats. Although renal insufficiency did not change gastric emptying (median %GR = 26.6 for the nephrectomized subgroup and 29.3 for the sham subgroup), LPS significantly retarded the gastric emptying of the sham and nephretomized groups (median %GR = 42.0 and 61.0, respectively), and was significantly greater (P<0.01) in the nephrectomized rats. We conclude that gastric emptying in animals suffering from moderate renal insufficiency is more sensitive to the action of LPS than in sham animals

Activity and functional interaction of alternative oxidase and uncoupling protein in mitochondria from tomato fruit

Cyanide-resistant alternative oxidase (AOX) is not limited to plant mitochondria and is widespread among several types of protists. The uncoupling protein (UCP) is much more widespread than previously believed, not only in tissues of higher animals but also in plants and in an amoeboid protozoan. The redox energy-dissipating pathway (AOX) and the proton electrochemical gradient energy-dissipating pathway (UCP) lead to the same final effect, i.e., a decrease in ATP synthesis and an increase in heat production. Studies with green tomato fruit mitochondria show that both proteins are present simultaneously in the membrane. This raises the question of a specific physiological role for each energy-dissipating system and of a possible functional connection between them (shared regulation). Linoleic acid, an abundant free fatty acid in plants which activates UCP, strongly inhibits cyanide-resistant respiration mediated by AOX. Moreover, studies of the evolution of AOX and UCP protein expression and of their activities during post-harvest ripening of tomato fruit show that AOX and plant UCP work sequentially: AOX activity decreases in early post-growing stages and UCP activity is decreased in late ripening stages. Electron partitioning between the alternative oxidase and the cytochrome pathway as well as H+ gradient partitioning between ATP synthase and UCP can be evaluated by the ADP/O method. This method facilitates description of the kinetics of energy-dissipating pathways and of ATP synthase when state 3 respiration is decreased by limitation of oxidizable substrate.

Structural, functional and immunological studies on a polymeric bacterial protein

The characterization of proteins from Brucella spp, the causative agent of brucellosis, has been the subject of intensive research. We have described an 18-kDa cytoplasmic protein of Brucella abortus and shown the potential usefulness of this protein as an antigen for the serologic diagnosis of brucellosis. The amino acid sequence of the protein showed a low but significant homology with that of lumazine synthases. Lumazine is an intermediate product in bacterial riboflavin biosynthesis. The recombinant form of the 18-kDa protein (expressed in E. coli) folds like the native Brucella protein and has lumazine-synthase enzymatic activity. Three-dimensional analysis by X-ray crystallography of the homolog Bacillus subtilis lumazine synthase has revealed that the enzyme forms an icosahedral capsid. Recombinant lumazine synthase from B. abortus was crystallized, diffracted X rays to 2.7-Å resolution at room temperature, and the structure successfully solved by molecular replacement procedures. The macromolecular assembly of the enzyme differs from that of the enzyme from B. subtilis. The Brucella enzyme remains pentameric (90 kDa) in its crystallographic form. Nonetheless, the active sites of the two enzymes are virtually identical at the structural level, indicating that inhibitors of these enzymes could be viable pharmaceuticals across a broad species range. We describe the structural reasons for the differences in their quaternary arrangement and also discuss the potential use of this protein as a target for the development of acellular vaccines.

The starch from Solanum lycocarpum St. Hill. fruit is not a hypoglycemic agent

We have investigated the hypoglycemic effect induced by the starch obtained from the unripe fruits of Solanum lycocarpum (Solanaceae). Per os administration of the starch (1000 or 2000 mg/kg, twice daily for 7 days, N = 6) did not change glycemia levels of nondiabetic female Swiss mice weighing 25-30 g. In streptozotocin-induced diabetic mice, similar treatment with the starch did not change the elevated glycemia 3 h after the last dose (diabetic treated with saline = 288 ± 17/309 ± 18; starch 1000 mg/kg = 295 ± 33; starch 2000 mg/kg = 258 ± 37; N = 5). In animals fasted for 15 h, per os administration of glucose (600 mg/kg) significantly increased glycemia 1 h later. Previous (-30 min) treatment of the animals with the starch (1000 or 2000 mg/kg; N = 5) did not change the increase of glycemia. Per os administration of the starch (1000 or 2000 mg kg-1 day-1, twice daily for 7 days) did not induce body weight gain or loss. The chemical analysis of the starch indicated the presence of glycoalkaloids, a finding that represents a reason for concern since many of these substances are generally toxic. In interviews with 56 diabetic patients, 29 medicinal plants were reported as useful in their treatment of diabetes and S. lycocarpum was the sixth most frequently mentioned. All patients interviewed reported that they also used insulin or oral hypoglycemic drugs. The results of the present study do not provide evidence for a hypoglycemic effect associated with the polysaccharide fraction of S. lycocarpum in either normal or hyperglycemic mice. These data demonstrate the need for adequate pharmacological investigation of the natural products widely used in folk medicine.

Identification and in vitro production of Lactobacillus antagonists from women with or without bacterial vaginosis

Lactobacilli isolated from the vaginal tract of women with and without bacterial vaginosis (BV) were identified and characterized for the production of antagonists. Bacterial samples were isolated from healthy women (N = 16), from patients with clinical complaints but without BV (N = 30), and from patients with BV (N = 32). Identification was performed using amplified ribosomal DNA restriction analysis. Production of antagonistic compounds was evaluated by the double-layer diffusion technique using Gram-positive (N = 9) and Gram-negative bacteria (N = 6) as well as yeast (N = 5) as indicator strains. Of a total of 147 isolates, 133 were identified as pertaining to the genus Lactobacillus. Lactobacillus crispatus was the species most frequently recovered, followed by L. johnsonii and L. jensenii. Statistical analysis showed that L. crispatus was more frequent in individuals without BV (P < 0.05). A higher production of antagonists was noted in L. crispatus isolates from healthy women (P < 0.05). More acidic local pH and higher H2O2 production by isolated lactobacilli from healthy women suggest these mechanisms as the possible cause of this antagonism. In conclusion, a significant correlation was detected between the presence and antagonistic properties of certain species of Lactobacillus and the clinical status of the patients.

Randomized clinical trial comparing the efficacy of the vaginal use of metronidazole with a Brazilian pepper tree (Schinus) extract for the treatment of bacterial vaginosis

A 7.4% vaginal extract of the Brazilian pepper tree (Schinus terebinthifolius Raddi) was compared with 0.75% vaginal metronidazole, both manufactured by the Hebron Laboratory, for the treatment of bacterial vaginosis, used at bedtime for 7 nights. The condition was diagnosed using the combined criteria of Amsel and Nugent in two groups of 140 and 137 women, aged between 18 and 40 years. Intention-to-treat analysis was performed. Women were excluded from the study if they presented delayed menstruation, were pregnant, were using or had used any topical or systemic medication, presented any other vaginal infections, presented hymen integrity, or if they reported any history suggestive of acute pelvic inflammatory disease. According to Amsel’s criteria separately, 29 patients (21.2%) treated with the extract and 87 (62.1%) treated with metronidazole were considered to be cured (P < 0.001). According to Nugent’s score separately, 19 women (13.9%) treated with the extract and 79 (56.4%) treated with metronidazole were considered to be cured (P < 0.001). Using the two criteria together, the so-called total cure was observed in 17 women (12.4%) treated with the extract and in 79 women (56.4%) treated with metronidazole (P < 0.001). In conclusion, the cure rate for bacterial vaginosis using a vaginal gel from a pepper tree extract was lower than the rate obtained with metronidazole gel, while side effects were infrequent and non-severe in both groups.

Typical and atypical enteropathogenic Escherichia coli bacterial translocation associated with tissue hypoperfusion in rats

Although enteropathogenic Escherichia coli (EPEC) are well-recognized diarrheal agents, their ability to translocate and cause extraintestinal alterations is not known. We investigated whether a typical EPEC (tEPEC) and an atypical EPEC (aEPEC) strain translocate and cause microcirculation injury under conditions of intestinal bacterial overgrowth. Bacterial translocation (BT) was induced in female Wistar-EPM rats (200-250 g) by oroduodenal catheterization and inoculation of 10 mL 10(10) colony forming unit (CFU)/mL, with the bacteria being confined between the duodenum and ileum with ligatures. After 2 h, mesenteric lymph nodes (MLN), liver and spleen were cultured for translocated bacteria and BT-related microcirculation changes were monitored in mesenteric and abdominal organs by intravital microscopy and laser Doppler flow, respectively. tEPEC (N = 11) and aEPEC (N = 11) were recovered from MLN (100%), spleen (36.4 and 45.5%), and liver (45.5 and 72.7%) of the animals, respectively. Recovery of the positive control E. coli R-6 (N = 6) was 100% for all compartments. Bacteria were not recovered from extraintestinal sites of controls inoculated with non-pathogenic E. coli strains HB101 (N = 6) and HS (N = 10), or saline. Mesenteric microcirculation injuries were detected with both EPEC strains, but only aEPEC was similar to E. coli R-6 with regard to systemic tissue hypoperfusion. In conclusion, overgrowth of certain aEPEC strains may lead to BT and impairment of the microcirculation in systemic organs.

Air blast freezing of fruit pulp models in commercial boxes: influence of preferential channels in the bed on freezing times estimating

The freezing times of fruit pulp models packed and conditioned in multi-layered boxes were evaluated under conditions similar to those employed commercially. Estimating the freezing time is a difficult practice due to the presence of significant voids in the boxes, whose influence may be analyzed by means of various methods. In this study, a procedure for estimating freezing time by using the models described in the literature was compared with experimental measurements by collecting time/temperature data. The following results show that the airflow through packages is a significant parameter for freezing time estimation. When the presence of preferential channels was considered, the predicted freezing time in the models could be 10% lower than the experimental values, depending on the method. The isotherms traced as a function of the location of the samples inside the boxes showed the displacement of the thermal center in relation to the geometric center of the product.

Postharvest respiratory activity and changes in some chemical constituents during maturation of yellow mombin (Spondias mombin) fruit

Mature fruit from the yellow mombin (Spondias mombin) was monitored for its respiration activity. Mature green fruit from the yellow mombin was stored in closed glass chambers and the concentration of oxygen and carbon dioxide at the end of a six hour respiration period was determined. At the same interval of time, the lid of the chamber was opened for air renewal. The increase in carbon dioxide and decrease in oxygen concentration demonstrated that the fruit was climacteric. The maximum liberation of CO2 54.2 mL Kg-1 h-1 and maximum absorption of O2 49.0 mL Kg-1 h-1 occurred 186 hours after the harvest which, obviously, represented the optimum fruit quality after the senescence process started. The respiratory quotient of fruit at a climacteric maximum was 1.11 representing the oxidation of carbohydrates. Total soluble solids increased from 9.1 °Brix (initial) to 13.7 °Brix (climacteric maximum) during maturation, while the total number of acids in the fruit decreased during maturation i.e. from 1.55% initially to 1.40% at pre-climacteric, 1.0% at climacteric maximum and 0.8% in the post-climacteric stage. A similar behaviour was observed in the case of ascorbic acid. There was a continuous decrease in chlorophyll and a continuous increase in the carotenoid content of fruit during maturation.

Bacterial ecology of tilapia fresh fillets and some factors that can influence their microbial quality

The purpose of research was to investigate the bacterial ecology of tilapia (Oreochromis niloticus) fresh fillets and some factors that can influence its microbial quality. Samples of fish cultivation water (n = 20), tilapia tegument and gut (n = 20) and fresh fillets (n = 20) were collected in an experimental tilapia aquaculture located in the city of Lavras, Minas Gerais, Brazil. Staphylococcus spp., Aeromonas spp., Enterococcus spp. and Enterobacteriaceae were quantified using selective plating. For the enumeration of Pseudomonas spp., the most probable number technique (MPN) was utilized. Bacterial colonies (n = 198) were identified by Gram strain and biochemical tests. Aeromonas spp., Pseudomonas spp., Enterococcus spp. and Enterobacteriaceae were found in the cultivation water (water from a fishpond cultivation), tegument, gut, and fresh fillets. Staphylococcus spp. was not isolated in the cultivation water. Salmonella spp. was not detected. The count variable of 10 to 10³ CFU or MPN.(g or mL)-1. Associated to freshwater tilapia fillet processing, there is a large variety of microorganisms related to foodborne illnesses and fish products deterioration.