Significant Population Differences Research Articles

Transanal haemorrhoidal artery echodoppler ligation and anopexy (THD) is effective for II and III degree haemorrhoids: a prospective multicentric study

We report a multicentric prospective study which aimed to evaluate Doppler-assisted ligation of the terminal haemorrhoidal arteries (THD) for II and III degree haemorrhoids. A total of 112 patients from five colorectal units, including 81 men, mean age 48 +/- 13 years, with II degree (39) and III degree (73) haemorrhoids were treated by Doppler-guided transanal de-arterialization and anopexy using a new device (THD). The mean operative time was 33.9 +/- 8.8 minutes, and the mean number of ligatures applied was 7.2 +/- 1.5. Postoperatively, 72% of patients did not need analgesics and the other 28% used nonsteroidal antiinflammatory drugs 1-3 times/day for less than 2 days. All the patients were operated as a day case. Early postoperative complications included haemorrhoidal thrombosis (2 patients), bleeding (1) treated by haemostatic suture, dysuria (6) and acute urinary retention (1). After a mean follow-up of 15.6 +/- 6.5 months (range 6-32), 2/105 (20.9%) patients complained of minor bleeding, while mild pain was still present in 4/51 patients (7.8%). There were no statistically significant differences in the sample population regarding the gender or stage of the disease. Tenesmus was cured in 15/17 patients, dyschaezia in 20/22 patients and mucous soiling in 10/10 patients. No new cases of altered defaecation or faecal incontinence were recorded. Overall, 85.7% of patients were cured and 7.1% improved. Residual haemorrhoids were treated by elastic band ligation in nine (8%) patients and by surgical excision in further five patients (4.5%). Doppler-assisted ligation of the terminal branches of the haemorrhoidal arteries for II and III degree haemorrhoids is highly effective and painless. Complications are few and the technique can be performed as a day case.

Does Low‐Dose Heparin Maintain Central Venous Access Device Patency?

A common problem that complicates use of central venous access devices (CVADs) is occlusion by thrombosis. Alteplase, a recombinant tissue plasminogen activator, is used to restore line patency when thrombosis occurs. Heparin flush is commonly used to prevent this complication, but the effectiveness of this practice is unclear. A recent heparin shortage allowed examination of heparin effectiveness in reducing CVAD thrombosis. A retrospective cohort study was performed by querying a pharmacy database for alteplase use for CVAD thrombosis in adult patients during periods when heparin flushes (10 units/mL) were used and when saline flushes were used instead because of a nationwide heparin shortage. The number of patients receiving alteplase, the number of doses administered, and the total amount of alteplase used were compared over 1-month intervals of heparin flush use and 1-month intervals of saline flush use. Patient days and critical care patient days were compared between these time intervals. Peripherally inserted central catheter (PICC) line placements and replacements between time periods of heparin and saline flush were also compared. Significant increases in the number of patients receiving alteplase (P = .04), the number of alteplase doses administered (P = .04), and total dose of alteplase used (P = .05) occurred during the heparin shortage. No significant differences in patient population were observed. The percentage of PICC line replacements also increased significantly (P < .05) when heparin was not available. Heparin flush (10 units/mL) decreases thrombotic occlusions of CVADs, resulting in decreased alteplase use and fewer PICC line replacements.

Comparative determination of HIV-1 co-receptor tropism by Enhanced Sensitivity Trofile, gp120 V3-loop RNA and DNA genotyping

BackgroundTrofile® is the prospectively validated HIV-1 tropism assay. Its use is limited by high costs, long turn-around time, and inability to test patients with very low or undetectable viremia. We aimed at assessing the efficiency of population genotypic assays based on gp120 V3-loop sequencing for the determination of tropism in plasma viral RNA and in whole-blood viral DNA. Contemporary and follow-up plasma and whole-blood samples from patients undergoing tropism testing via the enhanced sensitivity Trofile® (ESTA) were collected. Clinical and clonal geno2pheno[coreceptor] (G2P) models at 10% and at optimised 5.7% false positive rate cutoff were evaluated using viral DNA and RNA samples, compared against each other and ESTA, using Cohen's kappa, phylogenetic analysis, and area under the receiver operating characteristic (AUROC).ResultsBoth clinical and clonal G2P (with different false positive rates) showed good performances in predicting the ESTA outcome (for V3 RNA-based clinical G2P at 10% false positive rate AUROC = 0.83, sensitivity = 90%, specificity = 75%). The rate of agreement between DNA- and RNA-based clinical G2P was fair (kappa = 0.74, p < 0.0001), and DNA-based clinical G2P accurately predicted the plasma ESTA (AUROC = 0.86). Significant differences in the viral populations were detected when comparing inter/intra patient diversity of viral DNA with RNA sequences.ConclusionsPlasma HIV RNA or whole-blood HIV DNA V3-loop sequencing interpreted with clinical G2P is cheap and can be a good surrogate for ESTA. Although there may be differences among viral RNA and DNA populations in the same host, DNA-based G2P may be used as an indication of viral tropism in patients with undetectable plasma viremia.

Conformational disorder in 4-(5,5′-dibromo-2′-chloro-4,4′-bipyridyl-2-yl)benzaldehyde: role of π–π and halogen interactions

The crystal packing of the title compound, C(17)H(9)Br(2)ClN(2)O, is governed by strong pi-pi stacking, where molecules are tightly bound within infinite (100) planes; these planes interact mainly through non-optimal pi-pi stacking where arene rings are noticeably displaced from perfect overlap, and also through halogen-halogen interactions. The aldehyde group shows conformational disorder, with a significant population difference between the two conformers; this difference is rationalized by the energetic analysis of the crystal packing using the PIXEL method, which also allows a decomposition of intermolecular interaction energy into Coulombic, polarization, dispersion and repulsion contributions. Using such an analysis, it is found that the main reason for this unequal population of the two conformers in the crystal is two hydrogen bonds that are present only for the major conformer.

Human leukocyte antigen crossmatch testing is important for liver retransplantation

Although human leukocyte antigen (HLA) crossmatching is often thought to be unnecessary for liver transplants (LTs), we provide evidence that for retransplants, it is essential. Sera from 139 retransplant patients who had received livers from deceased donors were retrospectively analyzed with single antigen beads on a Luminex platform for HLA antibodies. Each patient received at least 2 transplants and was followed up for at least 6 months from the second LT, which was deemed to have failed if the patient had a third LT or died. Second LT survival was calculated from the date of the second LT to the date of the third LT or death. Our study cohort consisted of 118 adult patients (> or = 18 years old) as well as 21 pediatric patients (<18 years old). Class I HLA antibodies were associated with significantly poorer regraft survival in adults [survival differences of 21.3% (P = 0.046), 22.1% (P = 0.042), and 23.7% (P = 0.033) at 1, 3, and 5 years, respectively]; however, the presence of these antibodies was not associated with significant survival differences in the pediatric population. A univariate analysis of the effect of class I antibodies on second LT survival in adults showed a hazard ratio of 2.0 (95% confidence interval = 1.0-3.8, P = 0.028). Graft survival in patients with and without HLA antibodies or class II antibodies was similar. Because class I antibodies have a deleterious effect on liver regraft survival, crossmatch testing should be performed before liver retransplantation.

Drought management in sugarcane during pre-monsoon period

A Field experiment was conducted during spring season of 2005–06, 2006–07 and 2007–08 to work out the agronomic practice for drought management of sugarcane during pre monsoon period. The treatments consisted of four methods of planting i.e. 60cm row spacing, 90 cm row spacing, trench planting at 120 cm distance and pit planting and four drought management practices, viz., T1- Control (Recommended practices of the region), T2- soaking of setts in saturated lime water+ KCl and urea foliar spray each at 90,105 and 120 days after planting (DAP)+ trash mulching after 60DAP+ addition of FYM@ 10t/ha in furrows before planting, T3–T2+ additional potassium @ 60 kg K2O at first irrigation and T4–T2+ additional potassium @ 60 kg K2O/ha at 150–180 days after planting. Highest cane and sugar yield was recorded under pit planting on pooled basis followed by 60cm, trench and 90cm spacing. There was no significant difference in shoot population, millable canes and cane yield of drought management practices plots (T2, T3, T4) and normal irrigated plots (T1). Combination of different technologies, viz., soaking of setts in saturated lime water, application of FYM and foliar spray of KCl and Urea proved effective in managing the sugarcane under drought conditions. Highest gross returns, variable cost, net return and B: C ratio were recorded in pit planting followed by 60cm row spacing

Comprehending the Thermal Decomposition and Reconstruction Process of Sol−Gel MgAl Layered Double Hydroxides

The evolution of MgAl layered double hydroxides (LDHs), prepared by the sol−gel method, throughout the synthesis−crystallization−calcination−reconstruction process is thoroughly studied and compared to a coprecipitated analogue. X-ray diffraction and transmission electron microscopy provide useful information about crystallinity and morphology of the samples. It was found that crystal sizes do not necessarily decrease after calcination−reconstruction. Furthermore, aluminum coordination is studied by 27Al MAS nuclear magnetic resonance and by X-ray photoelectron spectroscopy (XPS), revealing significant differences in the relative populations of tetrahedral aluminum in the surface and the bulk of the calcined sol−gel LDHs. Magnesium and oxygen structural environments are also studied by XPS. A possible correlation between crystal sizes of calcined LDHs and the amount of O2− surface species is discussed. Differences in the structural environment of magnesium in the as-synthesized and the reconstructed LDH a...

Songbird chemosignals: volatile compounds in preen gland secretions vary among individuals, sexes, and populations

Chemical signaling has been documented in many animals, but its potential importance in avian species, particularly songbirds, has received far less attention. We tested whether volatile compounds in the preen oil of a songbird (Junco hyemalis) contain reliable information about individual identity, sex, or population of origin by repeated sampling from captive male and female juncos originating from 2 recently diverged junco populations in southern California. One of the populations recently colonized an urban environment; the other resides in a species-typical montane environment. The birds were field-caught as juveniles, housed under identical conditions, and fed the same diet for 10 months prior to sampling. We used capillary gas chromatography-mass spectrometry to quantify the relative abundance of 19 volatile compounds previously shown to vary seasonally in this species. We found individual repeatability as well as significant sex and population differences in volatile profiles. The persistence of population differences in a common environment suggests that preen oil chemistry likely has a genetic basis and may thus evolve rapidly in response to environmental change. These finding suggest that songbird preen oil odors have the potential to function as chemosignals associated with mate recognition or reproductive isolation.

Circulating subsets and CD4+CD25+ regulatory T cell function in chronic inflammatory demyelinating polyradiculoneuropathy

Chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) is an inflammatory disease of the peripheral nervous system that is probably autoimmune in origin. Different components of the adaptive and innate immunity may be responsible for the aberrant response towards nerve antigens. To investigate this, we examined lymphocyte subsets and regulatory T cell (Treg) function in the blood of CIDP patients, healthy controls (HC) and subjects with non-immune mediated neuropathies (other neuropathies, ON). We used flow cytometry to determine the frequency of monocytes, B cells, natural killer (NK) and NK-T cells, total and activated CD4+ and CD8+ T cells, effector memory and central memory CD4+ and CD8+ T cells, and CD4+CD25highFoxp3+ Tregs. Treg function was studied after polyclonal stimulation and antigen specific stimulation with myelin protein peptides in CIDP and HC. There was an increased frequency of monocytes (p = 0.02) and decreased frequency of NK cells (p = 0.02) in CIDP compared with HC but not ON. There were no significant differences in other populations. Treg function was impaired in CIDP compared to HC (p = 0.02), whilst T cell proliferation to myelin protein peptides before and after depletion of Tregs was not different between patients and controls. This study shows increased circulating monocytes and reduced NK cells in CIDP. Although Treg frequency was not altered, we confirm that Tregs display a defect of suppressive function. Myelin protein peptides were not the target of the altered peripheral regulation of the immune response. The mechanisms of peripheral immune tolerance in CIDP and their relevance to the pathogenesis deserve further exploration.

Comparative studies of microbial populations in the rumen, duodenum, ileum and faeces of lactating dairy cows

Understanding factors that influence the composition of microbial populations of the digestive system of dairy cattle will be key in regulating these populations to improve animal performance. Although rumen microbes are well studied, little is known of the dynamics and role of microbial populations in the small intestine of cows. Comparisons of fingerprints of microbial populations were used to investigate the effects of gastrointestinal (GI) segment and animal on community structure. Samples from four lactating dairy cows with ruminal, duodenal and ileal cannulae were collected. Terminal-restriction fragment length polymorphism (T-RFLP) comparisons of small subunit rRNA genes revealed differences in microbial populations between GI segments (P < 0.05). No significant differences in either methanogen populations or microbial community profiles between animals were observed. Quantitative PCR was used to assay relative changes in methanogen numbers compared to procaryote rRNA gene numbers, and direct microscopic counts were used to enumerate total procaryote numbers of the duodenal and ileal samples. T-RFLP comparisons illustrate significant changes in microbial diversity as digesta passes from one segment to another. Direct counts indicate that microbial numbers are reduced by eight orders of magnitude from the rumen, through the abomasum, and into the duodenum (from c. 10(12) to c. 3.6 x 10(4) cells per ml). Quantitative PCR analyses of rRNA genes indicate that methanogens are present in the duodenum and ileum. The contribution of microbial populations of the small intestine to the nutrition and health of cattle is seldom addressed but warrants further investigation.

Antimicrobial Activity of Commercial Concrete Sealant Against Salmonella Spp: A Model for Poultry Processing Plants

Salmonella is an important foodborne pathogen often associated with poultry and highly prevalent in poultry processing plants. The objective of this study was to determine the efficacy of a commercial grade concrete sealant (BioSealed for Concrete ) to prevent bacterial attachment, colonization and antimicrobial TM following treatments C and D as compared to treatments A and B. However, no significant differences (p>0.05) in the survival populations of Salmonella were observed between treatments A and B for all five strains tested and between treatments C and D for any of the strains tested. This indicates that BioSealed for Concrete proved to be a potent antimicrobial against multiple strains of Salmonella and can be used TM

Comparison of melanoblast expression patterns identifies distinct classes of genes.

A full understanding of transcriptional regulation requires integration of information obtained from multiple experimental datasets. These include datasets annotating gene expression within the context of an entire organism under normal and genetically perturbed conditions. Here we describe an expression dataset annotating pigment cell-expressed genes of the developing melanocyte and retinal pigmented epithelium lineages. Expression images are annotated and available at http://research.nhgri.nih.gov/manuscripts/Loftus/March2009/. Data are also summarized in a standardized manner using a universal melanoblast scoring scale that accounts for the embryonic location of cells and regional cell density. This approach allowed us to classify 14 pigment genes into four groupings classified by cell lineage expression, temporal-spatial context, and differential alteration in response to altered MITF and SOX10 status. Significant differences in regional populations were also observed across inbred strain backgrounds, highlighting the value of this approach to identify modifier allele influences on melanoblast number and distributions. This analysis revealed novel features of in vivo expression patterns that are not measurable by in vitro-based assays, providing data that in combination with genomic analyses will allow modeling of pigment cell gene expression in development and disease.

Efficacy of detergents and fresh produce disinfectants against microorganisms associated with mixed raw vegetables.

Efficacy of commercial detergent and disinfectants to eliminate microorganisms associated with fresh vegetables eaten raw in Iran, including radish, parsley, basil, coriander (cilantro), Allium porrum (leek), and peppermint were studied. The raw vegetables were subjected to a triple wash treatment of washing in tap water for mud removal, washing in water containing a detergent (dishwashing liquid) or disinfectant individually, and rinsing in tap water. The population of total mesophilic microbes on the surface of untreated vegetables ranged from 10(5) to 10(6) CFU/g. Washing in tap water or treatment with detergent (333 ppm for 10 min) or benzalkonium chloride (92 ppm for 15 min) reduced the total microbial count, most probable number (MPN) of coliforms, MPN of fecal coliforms, and MPN of fecal streptococci by about 1.2 to 2.3 log. No significant differences in microbial populations were found on vegetables after decontamination with tap water, detergent, or benzalkonium chloride (P > 0.05). Treatments with peracetic acid (100 ppm for 15 min) and hydrogen peroxide (133 ppm for 30 min) reduced the total mesophilic microbial counts by about 2.8 log. The microbial reductions with calcium hypochlorite (300 ppm for 15 min) and combined hydrogen peroxide and silver ion (133 ppm for 30 min) were significantly higher than those obtained after rinsing in tap water or after detergent or benzalkonium chloride wash (P < 0.05). Pretreatment with detergent slightly enhanced the efficacy of all decontamination treatments, but results were not significantly different from those obtained after individual application of disinfectants.

Genetic adaptation to extreme hypoxia:: Study of high-altitude pulmonary edema in a three-generation Han Chinese family

Organismal response to hypoxia is essential for critical regulation of erythropoiesis, other physiological functions, and survival. There is evidence of individual variation in response to hypoxia as some but not all of the affected individuals develop polycythemia, and or pulmonary and cerebral edema. A significant population difference in response to hypoxia exist as many highland Tibetan, Ethiopian, and Andean natives developed adaptive mechanisms to extreme hypoxia. A proportion of non-adapted individuals exposed to high altitude develop pulmonary edema (HAPE), pulmonary hypertension, cerebral edema, and extreme polycythemia. The isolation of causative gene(s) responsible for HAPE and other extreme hypoxia complications would provide a rational basis for specific targeted therapy of HAPE, allow its targeted prevention for at-risk populations, and clarify the pathophysiology of other hypoxic maladaptations. The only suggested genetic linkage among unrelated individuals with HAPE has been with endothelial nitric oxide synthase ( eNOS) gene. Here we describe a family with multiple members affected with HAPE in three generations. Families with multiple affected members with HAPE have not been described. We first ruled out linkage of HAPE with the eNOS gene. We then performed an analysis of the whole genome using high-density SNP arrays (Affymetrix v5.0) and, assuming a single gene causation of HAPE, ruled out linkage with 34 other candidate genes. Only the HIF2A haplotype was shared by individuals who exhibit the HAPE phenotype, and work on its possible causative role in HAPE is in progress. The small size of our family does not provide sufficient power for a conclusive analysis of linkage. We hope that collaboration with other investigators with access to more HAPE patients will lead to the identification of gene(s) responsible for HAPE and possibly other maladaptive hypoxic complications.

Assessing the immune state of dogs suffering from pituitary gland dependent hyperadrenocorticism by determining changes in peripheral lymphocyte subsets

In order to evaluate the immune state of dogs suffering from pituitary-dependent hyperadrenocorticism (PDH), peripheral lymphocyte subsets were examined. Twenty seven PDH dogs and eight healthy control dogs were used in the current study. Eight healthy dogs served as the control group. Twenty seven PDH dogs were categorized into 4 groups based on their post serum cortisol concentrations by ACTH stimulation test: 2-5, excellent control (n = 8); 5-20, fair control (n = 7); >20, poor control (n = 4); and untreated (n = 8). Cell counts were executed with white blood cells (WBC), lymphocytes, CD3(+) (T lymphocytes), CD4(+) (Helper T lymphocytes), CD8(+) (Cytotoxic T lymphocytes), CD21(+) (B lymphocytes) cells in addition to calculating CD4(+)/CD8(+) ratio. Results indicated a significant difference in lymphocyte numbers and lymphocyte subset populations (CD3(+), CD4(+), CD8(+), and CD21(+) cells) between PDH and control dogs. Moreover, comparison of the PDH groups (excellent control; fair control; poor control; untreated) demonstrated that all groups had a significant decrease in lymphocytes numbers (CD3(+), CD4(+) and CD21(+) cell counts) as compared to control group. Meanwhile, no significant differences were observed in WBC counts and CD4(+)/CD8(+) ratio between groups. Furthermore, lymphocyte subset distribution in excellent control PDH dogs without concurrent disease (n = 4) better resembled that of control dogs as compared to PDH dogs with concurrent disease (n = 4). PDH dogs may be suffering from an immuno-depressed state as evidenced by significant differences in lymphocyte subset populations. Furthermore, treatment of both PDH and concurrent disease might improve lymphocyte subset distribution.

Clinical role of regulatory T cell in intraductal papillary mucinous neoplasms

e15537 Background: Intraductal papillary mucinous neoplasms of pancreas (IPMNs) have malignant potential and exhibit a broad histologic spectrum, ranging from adenoma to invasive carcinoma. Recently, several investigators have reported that regulatory T cells (Foxp3+CD4+ T cells including CD4+CD25+ T cells and CD4+CD25- T cells) play important roles in anti-tumor immunity. We previously reported clinical role of increased populations of Foxp3+CD4+ T cells (Treg) in peripheral blood from advanced pancreatic cancer patients (Pancreas 2006). This study was conducted to clarify the clinical role of Treg for surveillance of IPMNs. Methods: Thirty-five patients with IPMNs (IPMC: n=8, IPMB: n=2, IPMA: n=5) and pancreas cancer (n=20) who underwent surgical resection were included in this study. Peripheral blood was withdrawn from IPMN patients (IPMC: n=5, IPMA: n=2) and PC patients (n=20), as well as healthy volunteer donors (n=20) as controls. The peripheral blood mononuclear cells (PBMCs) were subjected to FACScan analysis after labeling with anti-CD4, anti-CD25, and anti-Foxp3 antibodies. Immunohistochemical expression of Foxp3 (abcam: FOXP3 antibody, 236A/E7), Fascin (Dako: Fascin antibody, 55K-2) and TGF-β (Santa Cruz: TGF-β1 antibody, SC146) in main tumor was assessed in all IPMN patients. Results: The Treg population among the PBMCs was significantly increased in IPMC patients (median: 4.50%) and PC patients (median: 3.06%) compared with healthy donors (median: 1.30%). No significant difference in the Treg population among the PBMCs was observed between IPMC and PC patients. On the other hand, in IPMA patients (n=2), the Treg population among the PBMCs was equivarent to healthy donors (2.40 and 1.85% respectively). In immnohistochemical staining, the positive expression of Foxp3 was 87.5% (7 of 8) in IPMC patients, while in IPMA and IPMB patients, the expression of Foxp3 was not observed. The expression of Fascin and TGF-β was not correlated to malignant potential of IPMNs Conclusions: Increase of Foxp3+CD4+ T cells in the PBMCs is a new promising marker for malignant potential in IPMNs patients. No significant financial relationships to disclose.

Dialects in pygmy marmosets? Population variation in call structure

Population variation in primate vocal structure has been rarely observed. Here, we report significant population differences in the structure of two vocalizations in wild pygmy marmosets (Trills and J calls). We studied 14 groups of pygmy marmosets Callithrix (Cebuella) pygmaea pygmaea from five populations in northeastern Ecuador. We analyzed the acoustic structure of Trills and J calls recorded from two adult animals in each group through focal samples. Although individuals and groups within a population differed in call structure, we found consistent structural differences at a population level for Trills and J calls. Pair-wise comparisons for the two call types point to San Pablo and Amazoonico as the populations that differed the most, whereas Hormiga and Zancudococha showed the least differences. Discriminant function analysis indicates that calls from each population could be classified accurately at rates significantly above chance. Habitat acoustics, social factors and genetic drift may explain interpopulation vocal differences. This is the first evidence of within-subspecies vocal differences, or dialects, in wild populations of a neotropical primate species.

Modification of Multiple Sclerosis Phenotypes by African Ancestry at HLA.

In those with multiple sclerosis (MS), African American individuals have a more severe disease course, an older age at onset, and more often have clinical manifestations restricted to the optic nerves and spinal cord (opticospinal MS) than white persons. To determine whether genetic variation influences clinical MS patterns. Retrospective multicenter cohort study. Six hundred seventy-three African American and 717 white patients with MS. Patients with MS were genotyped for HLA-DRB1 and HLA-DQB1 alleles. The proportion of European ancestry at HLA was estimated by genotyping single-nucleotide polymorphisms with known significant frequency differences in West African and European populations. These genotypes were correlated with the opticospinal disease phenotype, disability measures, and age at onset. Subjects with DRB1*15 alleles were twice as likely to have typical MS rather than opticospinal MS (P = .001). Of the subjects with opticospinal MS or a history of recurrent transverse myelitis who were seropositive for anti-aquaporin 4 antibodies (approximately 5%), none carried DRB1*15 alleles (P = .008). Independently of DRB1*15, African ancestry at HLA correlated with disability as measured by the Multiple Sclerosis Severity Score (P < .001) and risk of cane dependency (hazard ratio, 1.36; P < .001); DRB1*15 alleles were associated with a 2.1-year earlier age at onset (P < .001). These data indicate that the role of HLA in MS is not limited to disease susceptibility but that genes embedded in this locus also influence clinical outcomes.

In Vitro Ultramorphological Assessment of Apoptosis Induced by Zerumbone on (HeLa )

Zerumbone (ZER), a potential anticancer compound, isolated from the fresh rhizomes of Zingiber zerumbet. In this investigation, the cytotoxic properties of ZER were evaluated, on cancer cells of human cervix (HeLa), breast and ovary, and normal cells of Chinese Hamster ovary, using MTT assay. Apoptogenic effects of ZER on HeLa were studied using fluorescence microscopy (AO/PI double staining), scanning and transmission electron microscopy (SEM and TEM), and colorimetric assay of the apoptosis promoter enzyme, caspase-3. The results of MTT assay showed that ZER has less effect on normal cells compared to cancer cells. The lowest IC50 of ZER was observed on HeLa cells. Cytological observations showed nuclear and chromatin condensation, cell shrinkage, multinucleation, abnormalities of mitochondrial cristae, membrane blebbing, holes, cytoplasmic extrusions and formation of apoptotic bodies as confirmed collectively by double staining of AO/PI, SEM and TEM. Statistical analysis (two-tailed t-test) of differential counting of 200 cells under fluorescence microscope revealed significant difference in apoptotic cells populations between treated and untreated HeLa cells. In addition, ZER has increased the cellular level of caspase-3 on the treated HeLa cells. It could be concluded that ZER was able to produce distinctive morphological features of cell death that corresponds to apoptosis.

Reduced-Risk Pest Management Programs for Eastern U.S. Apple and Peach Orchards: A 4-Year Regional Project

Studies were conducted from 2002 to 2005 to determine the effectiveness of reduced-risk (RR) tactics for managing key pests in 50 commercial apple orchards (114 ha) in Michigan, North Carolina, New York, Pennsylvania, Virginia, and West Virginia; and 20 peach orchards (190 ha) in Michigan, New Jersey, Pennsylvania, and West Virginia. At each apple site, a block of up to 5 ha received a seasonal program of selective RR and organophosphate-replacement insecticides, with or without pheromones for mating disruption of key lepidopteran pests of apple (codling moth, oriental fruit moth, and obliquebanded leafroller) and peach (oriental fruit moth, lesser peachtree borer, and peachtree borer). A comparison block at each site with the same varieties and tree training was managed using each grower's standard program of conventional insecticides (STD). Pheromone traps for lepidopteran species were hung in all plots and monitored weekly. Foliar samples were taken during the season to estimate phytophagous and predator mite densities. Red sphere traps baited with fruit volatiles were used to monitor apple maggot adults in apple orchards. Fruits were inspected for insect damage at harvest, and graded according to USDA standards. Partial budget analysis was used to assess the net profitability of RR programs to produce apples and peaches for their intended market in each state. Fruit damage at harvest caused by direct fruit pests was generally low across all blocks and treatments. There were no statistically significant differences in fruit damage or mite populations between the RR blocks, with or without pheromones, and the growers' standards. Insecticide use patterns in the RR plots represented up to 88 and 78% reduction in the amount of active ingredient applied per hectare, and an 85 and 77% decrease in their Environmental Impact Quotient for apples and peaches, respectively. However, RR programs were more expensive and generally less profitable compared with growers' standard programs. Regression analysis estimated that RR apple programs with and without mating disruption were on average &dollar;465 and &dollar;144/ha more expensive, and &dollar;544 and &dollar;159/ha less profitable, respectively, compared with standard programs. RR+MD programs for peaches cost an average &dollar;314/ha more and returned about &dollar;284/ha less than STD peach programs.