Seasonal Population Dynamics of Megacopta cribraria (Hemiptera: Plataspidae) in Kudzu and Soybean, and Implication for Insecticidal Management in Soybean.

Megacopta cribraria (F.), an invasive species introduced from Asia in 2009, is now prolific in the southeastern United States. Megacopta cribraria develops primarily on kudzu and soybean completing two generations. It is not well understood how this economic pest is affected by changes in geographic distribution in the United States or how population levels have changed since its establishment. The effect of insecticide application timing on field populations of M. cribraria is not well documented. These studies seek to understand how population dynamics of M. cribraria vary with geographic regions in Georgia. Effect of application timing on populations throughout the growing season was also examined. Weekly from 2012 to 2013, all life stages were enumerated from kudzu and soybean environments at several locations throughout Georgia from sweeps samples and flight intercept captures. Coordinates were recorded for locations, and classified as belonging to the Piedmont or Coastal Plain region of the state. Single spray trials were conducted from 2011-2014, and applications were made to soybean at intervals throughout the season. From 2012 to 2015, two kudzu patches near Griffin, GA, were monitored to detect population changes. Differences in population dynamics from locations around the state were found, but no clear effect of latitude, longitude, or region was observed. Insecticide applications applied in July suppressed nymph populations significantly better than treatments made earlier or later. Megacopta cribraria populations declined in 2014 and 2015 compared with 2012 and 2013. These studies provide the critical information for M. cribraria management in soybean in the southeastern United States.

Aberrant expression of chromogranin A, miR-146a, and miR-146b-5p in prostate structures with focally disrupted basal cell layers: an early sign of invasion and hormone-refractory cancer?

Our recent studies have suggested that prostate tumor invasion is triggered by autoimmunoreactions induced focal basal cell layer disruptions (FBCLD) that selectively favor monoclonal proliferation of the overlying progenitors or of a biologically more aggressive cell clone. As circulating chromogranin-A (CgA) levels are found to correlate with tumor progression and the status of hormone refractoriness, our current study attempted to assess whether CgA-positive cells would be preferentially distributed in epithelial structures with FBCLD. Paraffin-embedded specimens from 50 patients with organ-confined prostate cancer were subjected to double immunohistochemical analysis with monoclonal antibodies to basal cells and CgA. From each case, 3-5 randomly selected fields were digitally photographed and the photos were magnified 400% and the numbers of CgA-positive cells in epithelial structures with non-disrupted, focally disrupted, and lost basal cell layer were separately counted. The averaged number of cell for each category was statistically compared with the Pearson's Chi-square test. In addition, morphologically similar structures with and without CgA-positive cell clusters were microdissected from four selected cases and subjected to a comparison of differential micro-RNA expression levels. Our study revealed that, although isolated CgA-positive cells were seen in both the basal cell layer and the luminal cell population in all cases, only 8 cases (16%) harbored large clusters of CgA-positive cells that were concentrated in a given area, in which all or nearly all cells appeared to share a similar morphological and immunohistochemical profile. Microdissected epithelial structures with CgA-positive cell clusters exhibited a more than 5- and 7-fold lower expression of miR-146a and miR-146b-5p than their CgA-negative counterparts. As focal basal cell layer disruptions and the reduction or loss of miR-146a and miR-146b-5p has been documented to correlate with prostate tumor invasion and hormone refractoriness, our findings suggest that aberrant CgA expression in epithelial structures with FBCLD may represent an early sign of these events.

Activity of bifenthrin, chlorfenapyr, fipronil, and thiamethoxam against red imported fire ants (Hymenoptera: Formicidae).

Bifenthrin, chlorfenapyr, fipronil, and thiamethoxam were evaluated for activity against the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae). Mobility impairment and lethal times were determined after topical treatments. Both immobilization and mortality occurred most quickly with bifenthrin, followed by thiamethoxam, chlorfenapyr, and fipronil. Mortality due to horizontal exposure was evaluated at 10, 20, or 30 degrees C, with three ratios of topically treated donor ant corpses to live recipients (5, 10, or 20% donors). Bifenthrin had the greatest horizontal activity of the chemicals tested. For chlorfenapyr, the only treatments having higher mortality than controls were the highest percentage donors at either 10 or 30 degrees C. Horizontal activity of fipronil was temperature dependent only with the highest proportion of donors and was lower than that ofbifenthrin but higher than that of chlorfenapyr or thiamethoxam. Mean mortality due to thiamethoxam was similar to that with chlorfenapyr. Significant mortality occurred in all of the 20 and 30 degrees C thiamethoxam treatments, but none of the 10 degrees C treatments. Effectiveness as a barrier was evaluated by providing a choice between bridges treated with insecticide or water. Although bifenthrin did not provide an impenetrable barrier, it was the only treatment having fewer ants than its paired control bridge. Mortality data suggest that a reduction in recruitment rather than repellency account for this result.

Activity of bifenthrin, chlorfenapyr, fipronil, and thiamethoxam against Argentine ants (Hymenoptera: Formicidae).

Bifenthrin, chlorfenapyr, fipronil, and thiamethoxam were evaluated for activity against the Argentine ant, Linepithema humile (Mayr) (Hymenoptera: Formicidae). Mobility impairment and lethal times were determined after topical treatments. Ants were immobilized most quickly by bifenthrin, followed by chlorfenapyr and thiamethoxam. After 2 h, the number of fipronil-treated ants unable to walk out of test arenas did not differ from control ants. Median lethal time (LT50) after topical treatment was lowest in the bifenthrin treatment, followed by thiamethoxam, chlorfenapyr, and then fipronil. Mortality due to horizontal exposure was evaluated at 10, 20, or 30 degrees C, with topically treated ant corpses serving as donors. There was low to moderate horizontal activity in bifenthrin and chlorfenapyr treatments, with no temperature effect in bifenthrin treatments and a positive temperature effect in chlorfenapyr treatments. Mortality in the fipronil treatments was highest and was positively correlated with temperature. Thiamethoxam treatments did not differ from controls at 10 degrees C, but mortality increased with temperature. To evaluate contact activity, either all of 20% of the ants in a cohort were exposed to insecticide-treated pine needles. In both tests, mortality was highest in fipronil and bifenthrin treatments, followed by thiamethoxam, with lowest mortality in chlorfenapyr treatments. Effectiveness as a barrier was evaluated by providing a choice between bridges treated with insecticide or water. Although bifenthrin did not provide an impenetrable barrier, it was the only treatment having fewer ants than its paired control. Mortality data suggest that lack of recruitment rather than repellency account for this result.

Sialic acid-specific lectin from Tritrichomonas foetus.

A novel sialic acid-specific lectin (TFL) was isolated from Tritrichomonas foetus culture supernatant and purified by erythrocyte adsorption followed by fetuin-agarose affinity chromatography. According to gel filtration TFL is a protein of 728 kDa, different from the two sialidases of 853 and 254 kDa, secreted by T. foetus into the medium. The lectin is formed by multimeric complexes of 66 kDa subunit according to SDS-PAGE under reducing conditions. TFL is glycosylated with 4.2% of carbohydrates, half of which is represented by glucose. The lectin reacts equally with N-acetyl and N-glycolyl neuraminic acid, free, in alpha2,3- or alpha2,6-linkage. TFL has 7-fold weaker affinity to alpha2,8-linked N-acetylneuraminic acid (Neu5Ac) in colominic acid. Horse erythrocytes containing 4-O-acetyl Neu5Ac are agglutinated equally as compared to the human cells. TFL affinity to 9-O-acetyl Neu5Ac is 4-fold weaker as documented by hemagglutination inhibition with de-O-acetylated bovine submaxillary mucin, and ovine submaxillary mucin. A panel of mono- and oligosaccharides other than Neu5Ac do not inhibit TFL activity at 200 mM. The lectin does not require bivalent cations for activity, shows optimal reactivity at neutral pH and is stable at 4 degrees C. Anti-TFL antibodies identify membrane positivity on T. foetus, suggesting that the lectin functions in adhesion of the parasites. These findings, together with good stability and immunogenicity, make TFL a prospective candidate for further studies, especially in searching for efficient diagnostics and prevention of bovine trichomoniasis.

Clinical evaluation of hydroxyapatite-coated titanium plasma-sprayed and titanium plasma-sprayed cylinder dental implants: a preliminary report.

OBJECTIVE: The purpose of this article was to present the preliminary results of a prospective clinical trial comparing titanium plasma-sprayed versus hydroxyapatite-coated titanium plasma-sprayed cylinder (press fit) implants in different regions of the mouth. STUDY DESIGN: Sixty-five subjects met the inclusion requirements. Surgery was done in two phases by four experienced surgeons. Implant placement and abutment connection were separated by 3 to 4 months in the mandible, 6 to 7 months in the maxilla. Patients were assigned to either titanium plasma-sprayed or hydroxyapatite-coated implants on the day of surgery. Implant placement was not stratified for the region of the jaws. Outcome assessment was failure (loss) of an implant before or within 3 months of second phase surgery. RESULTS: Three hundred fifty-two implants equally distributed between titanium plasma-sprayed and hydroxyapatite-coated titanium plasma-sprayed implants were placed in four different sites; anterior maxilla, posterior maxilla, anterior mandible, and posterior mandible. There were a total of 15 failures (4.26%). Overall, titanium plasma-sprayed implants showed a higher but not significant failure rate compared with hydroxyapatite-coated implants (p = 0.06). Although not statistically significant, we believe that a smoking history played an important role in the failure of implants. CONCLUSION: This study suggests that an hydroxyapatite-coating of an implant allows superior initial integration when compared with a titanium plasma-sprayed surface.

Histochemical localization of sialylated glycoconjugates with Tritrichomonas mobilensis lectin (TLM).

A new sialic acid-specific lectin from the colonic parasite of squirrel monkeys Tritrichomonas mobilensis (TML) was tested on human and mouse tissues for histochemical staining properties. There were no substantial differences in reactivity between frozen and formalin fixed paraffin sections. TML staining was blocked by preincubation with sialic acid or by sialidase digestion. TML/anti-TML antibody histochemistry was identical with the TML-gold technique. The staining pattern was not blood group dependent. TML stained strongly the luminal membranes of normal vascular endothelium as well as endothelial neoplasms. Lymphatic vessels and capillaries of kidney glomeruli and lung alveolar septi were negative or only slightly positive. In parenchymatous organs luminal membrane positivity was dominant, preferably of cells lining ducts. Weak fine-granular cytoplasmic and basolateral membrane staining was also observed. Umbrella cells in transitional epithelium and basal layers of squamous epithelia showed strong reactivity with cell membranes. Mucin in respiratory epithelium was positive whereas gastrointestinal mucins failed to stain uniformly. Erythrocytes and most white blood cell types showed distinct membrane positivity. Acetylation or alkaline O-deacetylation of tissue sections did not substantially change TML reactivity. Oxidation, however, completely blocked TML staining except for respiratory epithelium and colonic mucin.

Sialic acid expression in normal and diseased human kidney.

Three sialic acid-specific lectins, Tritrichomonas mobilensis (TML) (without linkage preference), Maackia amurensis leukoagglutinin (MAL) (alpha2,3) and Sambucus nigra agglutinin (alpha2,6 linkage-specific), were used for detection of sialylated glycoconjugates in normal and diseased human kidneys. Normal kidneys demonstrated strong podocyte positivity of alpha2,3 linked sialic acid and weaker sialic acid expression on capillary endothelium, which was alpha2,6 linked. Renal biopsies (45) representing a variety of diseases, e.g. minimal change disease, membranous, membranoproliferative glomerulonephritis, tubulointerstitial nephrosis or diabetic glomerulopathy showed increased sialic acid expression in glomerular capillaries, Bowman's capsule epithelium and on podocytes. In several different kidney diseases the glomerular endothelium expressed also alpha2,3 linked sialic acid along with increased TML-positivity of epithelial cells related to alpha2,6 linked sialic acid. No difference in sialic acid linkage expression was observed in the tubules, which expressed putative alpha2,6 linked sialic acid on the luminal surface of cells distal to the descending limb of Henle's loop. The study did not show changes characteristic of specific diseases. Rather, increased sialic acid expression on glomerular endothelium and podocytes was linked to a variety of pathological changes. It is assumed that changes in sialysation of glycoconjugates in the glomeruli represent nonspecific changes and do not reflect fundamental pathogenetic features of renal diseases.

[Histochemistry of a new Tritrichomonas mobilensis lectin with sialic acid specificity]. / Histochémia nového lektínu Tritrichomonas mobilensis so specifictou pre kyselinu sialovú.

BACKGROUND: Histochemistry with lectins highly specific for one sugar represent a sensitive and accurate method for the localization of glycoproteins in tissues. OBJECTIVES: The paper evaluates the possibilities of histochemical applications of the novel lectin from Tritrichomonas mobilensis (TML) with specificity for sialic acid. METHODS: TML purified by affinity chromatography was used for anti-TML monoclonal antibody production and tested with the use of avidin-biotin peroxidase technique. RESULTS: In human biopsy material from different organs and tissues, TML showed strong membrane reactivity with vascular endothelium with the exception of glomerular and pulmonary capillaries and sinuses in the spleen and the bone marrow. Luminal membrane positivity dominated in glandular epithelia, especially in the lining of ducts. In squamous epithelia, the membrane positivity was present in cells of the basal layer, in transitional epithelium also in umbrella cells. Mucus in respiratory tract was positive, in gastrointestinal tract the positivity was irregular. Nerve tissue and endocrine glands were mostly negative. CONCLUSIONS: The lectin from tritrichomonas mobilensis can be considered a perspective reagent for identification of sialylated glycosubstances based on its high specificity for one sugar--the sialic acid and excellent results in histochemistry. (Fig. 7, Ref. 29.)

Purification and characterization of a sialic acid-specific lectin from Tritrichomonas mobilensis.

New sialic acid-specific lectin has been isolated from culture supernatant of the protozoan Tritrichomonas mobilensis. It was purified by adsorption by erythrocytes or bovine submaxillary gland mucin (BSM)-Sepharose affinity chromatography. The T. mobilensis lectin (TML) does not require bivalent cations for activity and agglutinates all human erythrocytes. The lectin forms multimeric complexes with molecular mass 556 and 491 kDa as determined by size-exclusion chromatography. SDS/PAGE under reducing conditions disclosed a large band of 343 kDa and three bands of 246, 265 and 286 kDa which, after denaturation with urea, were split into three subunits of 56, 61 and 66 kDa; under non-reducing conditions there were two bands, of 360 and 260 kDa. Western blots performed with anti-TML monoclonal antibodies revealed bands identical with those in the silver-stained gels, suggesting homogeneity of the BSM -Sepharose-purified lectin. TML is a highly glycosylated protein with approx. 8% of N-linked glycosides found by protein-N-glycanase F treatment; the total amount of saccharides revealed by chemical deglycosylation was 20%. Haemagglutination-inhibition studies documented exclusive specificity for sialic acid (NeuAc). Both (alpha 2-->6)- and (alpha 2-->3)-linked and free NeuAc were eight times more potent inhibitors than N-glycolylneuraminic acid. The lectin does not require O-acetyl groups on NeuAc for recognition. A spectrum of mono- and oligo-saccharides other than sialic acid had no inhibitory effect at 200 mM. Anti-TML monoclonal antibodies strongly inhibited the lectin activity. TML was stable at temperatures below 4 degrees C and lyophilized with 3% (w/w) glycerol.

Effect of environmental stress and surface treatment on resin-to-metal bonding.

This study compares the bond strength and durability of three metal surface treatments subjected to two types of environmental stress for both short- and long-term exposures. The luting resins Panavia and Comspan were applied to alumina-blasted, non-beryllium, nickel-chromium alloy coupons. Metal surface treatments consisted of either microscopic roughening by electrochemical etching, or one of two types of adhesives: a silanated silica coating (Silicoating) or a phosphate ester monomer (a component in the Panavia liquid). Shear bond strength was determined following short- or long-term exposure to either thermocycling in 6-60 degrees C water (2,672 cycles/7 days or 10,584 cycles/42 days) or storage in 37 degrees C water (7 or 42 days). Three-way ANOVA showed that both the type of environmental stress and the exposure time affected the bond strength of electroetched surfaces, but that only exposure time affected the two chemical adhesives (P < 0.05), regardless of the environmental stress used. In the short-term, the silica/silane coated surfaces produced and maintained the higher shear bond strengths (15.9 +/- 2.3 MPa). However, after 42 days the silica/silane bonds decreased 30% (to 11.3 +/- 2.2 MPa), while the phosphate ester bonds were essentially unchanged (11.4 +/- 3.0 at 4 days, 10.4 +/- 2.2 MPa at 42 days). Electroetched bonds were the weakest and decreased by 18% between 7 and 42 days in water (8.8 +/- 1.2 to 7.2 +/- 3.0 MPa) and 27% after 42 days of thermocycling (7.2 +/- 2.8 to 5.3 +/- 1.8 MPa).

Contact-independent cytotoxicity of Trichomonas vaginalis.

OBJECTIVE: To test the dependency of haemolytic and cytocidal manifestations of pathogenicity of Trichomonas vaginalis on direct contact between the target cells and the organism. TEST ORGANISM: T vaginalis strain Baltimore 42. DESIGN: Haemolysis in the presence of live T vaginalis and of its filter-sterilised metabolic products was compared. The dependence of haemolytic and cytocidal effects on retention of low pH of metabolic products of the organism was demonstrated by parallel titrations of sterile filtrates in normal saline and in phosphate buffered saline (PBS) pH 7.0. RESULTS: Near complete lysis was obtained when erythrocytes mixed with T vaginalis were incubated for 1 h at 37 degrees C in saline containing 1% glucose. The same degree of haemolysis was present in filter-sterilised glucose-saline in which the organism was incubated (1 h/37 degrees C) before erythrocytes were added and incubated under the same conditions as in the mixture with the organism. The degree of haemolysis in filtrates was dependent on retention of low pH (below 5.0) of the suspending fluid in which the organism alone was incubated. Dilution of filtrates in PBS, as opposed to normal saline, abolished or diminished the haemolytic effect. Presence of glucose (energy source) in the saline during incubation of the organism had a pronounced enhancing effect. The production of haemolytic metabolites was temperature dependent, whereas the haemolytic process per se was not. The effect was not an exclusive property of T vaginalis since it was also demonstrated with other trichomonads. The same filtrates applied to tissue culture exerted cytocidal effect strikingly similar to that observed in the haemolysis experiments. CONCLUSION: Neither haemolytic nor cytocidal effect of T vaginalis was contact-dependent.

Fetal prostate growth and development.

Normal fetal and newborn prostates were studied to evaluate growth patterns, histogenesis, and secretory activity. Whole cross-sectioned prostates harvested from 107 necropsies of fetuses and newborns ages 20 weeks gestation to 1 month of age were used. Development of the prostate occurred in three stages: bud stage (20-30 weeks gestation), bud-tubule stage (31-36 weeks gestation), and acinotubular stage (37-42 weeks gestation). Squamous metaplasia often appeared in the urethra, utricle, and primary lobular ducts, and occasional microcysts were noted. PAS and alcian blue-PAS positive secretion were present in 65% of the specimens by 20-30 weeks gestation and in over 87% of the specimens by 37 or more weeks gestation. Secretory activity was most prominent in the lateral regions of the peripheral zone. Prostate-specific antigen was only weakly detected throughout the prenatal period and was not related to secretory activity as evidenced by the PAS technique.

Acquisition and retention of viruses by Trichomonas vaginalis.

Recently described occurrence of virus-like particles (VLP) in some strains of Trichomonas vaginalis suggests the possibility that the pathogenic significance of this organism may be broadened by its potential for viral transmission. Inasmuch as neither the source nor the host range of the VLP are known, any hazard which they may present for man cannot be estimated. A model has been established for the study of acquisition of known human viruses by T vaginalis. Tissue cultures were infected with two reoviruses and a fresh isolate of genital herpes simplex virus (HSV). A squirrel monkey reovirus was also included in the study. T vaginalis was inoculated into the virus cultures three days later. The progress of virus acquisition by the trichomonads was monitored by transmission electron microscopy and by culture. Virus-containing cell fragments were engulfed by trichomonads and internalised in vacuoles. After digestion of cellular debris only virus particle aggregates were retained. Viable reoviruses were recovered from the trichomonads for nine days, and HSV for six days. The results suggest the possibility of transmission of at least some viruses by T vaginalis.