Estimation of genetic structure of a Mycosphaerella musicola population using inter-simple sequence repeat markers.

Among the diseases affecting banana (Musa sp), yellow Sigatoka, caused by the fungal pathogen Mycosphaerella musicola Leach, is considered one of the most important in Brazil, causing losses throughout the year. Understanding the genetic structure of pathogen populations will provide insight into the life history of pathogens, including the evolutionary processes occurring in agrosystems. Tools for estimating the possible emergence of pathogen variants with altered pathogenicity, virulence, or aggressiveness, as well as resistance to systemic fungicides, can also be developed from such data. The objective of this study was to analyze the genetic diversity and population genetics of M. musicola in the main banana-producing regions in Brazil. A total of 83 isolates collected from different banana cultivars in the Brazilian states of Bahia, Rio Grande do Norte, and Minas Gerais were evaluated using inter-simple sequence repeat markers. High variability was detected between the isolates, and 85.5% of the haplotypes were singletons in the populations. The highest source of genetic diversity (97.22%) was attributed to variations within populations. Bayesian cluster analysis revealed the presence of 2 probable ancestral groups, however, showed no relationship to population structure in terms of collection site, state of origin, or cultivar. Similarly, we detected noevidence of genetic recombination between individuals within different states, indicating that asexual cycles play a major role in M. musicola reproduction and that long-distance dispersal of the pathogen is the main factor contributing to the lack of population structure in the fungus.

New microsatellite markers for bananas (Musa spp).

Thirty-four microsatellite markers (SSRs) were identified in EST and BAC clones from Musa acuminata burmannicoides var. Calcutta 4 and validated in 22 Musa genotypes from the Banana Germplasm Bank of Embrapa-CNPMF, which includes wild and improved diploids. The number of alleles per locus ranged from 2 to 14. The markers were considered highly informative based on their polymorphism information content values; more than 50% were above 0.5. These SSRs will be useful for banana breeding programs, for studies of genetic diversity, germplasm characterization and selection, development of saturated genetic linkage maps, and marker assisted selection.

Characterization of Aspergillus flavus strains from Brazilian Brazil nuts and cashew by RAPD and ribosomal DNA analysis.

AIMS: The aim of this study was to determine the genetic variability in Aspergillus flavus populations from Brazil nut and cashew and develop a polymerase chain reaction (PCR) detection method. METHODS AND RESULTS: Chomatography analysis of 48 isolates identified 36 as aflatoxigenic (75%). One hundred and forty-one DNA bands were generated with 11 random amplified polymorphic DNA (RAPD) primers and analysed via unweighted pair group analysis, using arithmetic means (UPGMA). Isolates grouped according to host, with differentiation of those from A. occidentale also according to geographical origin. Aspergillus flavus-specific PCR primers ASPITSF2 and ASPITSR3 were designed from ribosomal DNA internal transcribed spacers (ITS 1 and 2), and an internal amplification control was developed, to prevent false negative results. Specificity to only A. flavus was confirmed against DNA from additional aspergilli and other fungi. CONCLUSIONS: RAPD-based characterization differentiated isolates according to plant host. The PCR primer pair developed showed specificity to A. flavus, with a detection limit of 10 fg. SIGNIFICANCE AND IMPACT OF THE STUDY: Genetic variability observed in A. flavus isolates from two Brazilian agroecosystems suggested reproductive isolation. The PCR detection method developed for A. flavus represents progress towards multiplex PCR detection of aflatoxigenic and nonaflatoxigenic strains in Hazard Analysis Critical Control Point systems.

The emotional problems of patients with bladder cancer.

In recent years the quality of survival has become a more dominant factor in our treatment and management of patients with cancer. As a result, it has become necessary to think in terms of a multidisciplinary approach to total rehabilitation. A part of this rehabilitation is of necessity determined by the emotional responses that patients exhibit to their disease and to its treatment. A useful model that has been applied to patients with other diseases and to patients with other types of cancer seems equally applicable to patients with bladder cancer. In my work with cancer patients I became aware that, after the initial fear of death was dealt with, the patients all responded to their dilemma with stereotyped and predictable responses (emotional patterns of behavior). It was apparent that these responses were determined by the degree to which the patient's life style was altered by the disease and/or treatment.

Expression of S9 and actin CyIIa mRNAs reveals dorso-ventral polarity and mesodermal sublineages in the vegetal plate of the sea urchin embryo.

We have used whole amount in situ hybridization to analyze the patterns of expression of two genes, S9 and actin CyIIa, during the development of the sea urchin, Strongylocentrotus purpuratus. We demonstrate that at the late blastula stage, these two mRNAs are expressed specifically by cells of the vegetal plate. Their domains of expression, however, are different. S9 mRNA is broadly distributed within most of the vegetal plate except for the central region, while CyIIa expression is restricted to a population of 10-15 cells in the ventral region of the plate. S9-expressing secondary mesenchyme cells (SMCs) migrate from the vegetal plate into the blastocoel early in gastrulation and later populate the dorsal ectoderm. The numbers, morphology, and migratory behavior of these cells strongly suggest that they are pigment cells. Throughout gastrulation, CyIIa mRNA is expressed by a population of presumptive SMCs at the ventral aspect of the archenteron tip. The pattern of expression of this mRNA is dynamic, however, and by the early pluteus stage, CyIIa mRNA accumulates in primary mesenchyme cells (PMCs), SMCs, and endodermal cells of the gut. When embryos are treated with NiCl2, a compound that has been shown to ventralize other embryonic tissues, CyIIa mRNA is expressed by an increased number of cells in the vegetal plate in a radially symmetrical pattern. The spatial pattern of CyIIa expression provides the first direct molecular evidence that the vegetal plate is polarized along the dorso-ventral (D-V) axis of the embryo. This gene product should be a valuable marker in future studies of D-V axis specification, as it can be detected at earlier developmental stages than existing molecular markers of this axis. Our observations show that the vegetal plate consists of subterritories of gene expression, and provide further support for the view that diversification of the presumptive, non-skeletogenic mesoderm begins prior to the onset of invagination.

Evidence for spread of the human immunodeficiency virus epidemic into low prevalence areas of the United States.

Reports of an increased proportion of AIDS cases occurring in small and medium-sized cities suggest that the HIV epidemic may be spreading into locations that were previously characterized by their low HIV antibody prevalences. Studying the question of the geographic spread of the HIV infection epidemic (rather than the AIDS epidemic) has been difficult largely because most serial seroprevalence data have been gathered from cohorts of high risk individuals (e.g., homosexual/bisexual cohorts) in New York City, San Francisco, and other geographically circumscribed areas. The U.S. military applicant HIV screening data were used in the current report to examine rates and 24 month temporal trends in geographic areas characterized by their HIV endemicities. The data examined concern the seven most populous states and four hyperendemic metropolitan areas located within those states (New York City, Miami, Houston, and San Francisco). In the nonepidemic regions, seroprevalence rates increased among black and white applicants. In the four epidemic urban areas, only young black applicants had higher HIV seroprevalence rates during the second 12 month period. Six of the seven nonepidemic regions had positive HIV seroprevalence trends, and these trends were significant in Florida, California, Texas, Illinois, and Ohio. The increases in these regions were greater for young blacks (30% excess for year 2 vs. year 1) compared to young whites (12% excess for year 2 vs. year 1). These data provide evidence of birth year specific increases in seroprevalence over time occurring in presumed low HIV prevalence areas. These increases cannot be due to, but are observed in spite of, biases associated with increasing self-selection over time.

Tracking the spread of the HIV infection epidemic among young adults in the United States: results of the first four years of screening among civilian applicants for U.S. military service.

Because the period from infection to clinically apparent disease is long, variable, and changing as new therapies are developed and applied, AIDS data are inadequate for tracking current values of critical parameters of HIV infection epidemics: prevalence of infection, rate of acquisition of new infections (incidence rate), and direction and rate of change of infection incidence over time (acceleration). These "vital signs" of infection epidemics can be tracked using serial cross-sectional seroprevalence data, however. From October 1985 through September 1989, more than 2.3 million applicants for U.S. military service were screened for antibody to HIV. The overall seroprevalence was 1.31 per 1,000 (3,014/2,300,675). Seroprevalences were highest near urban centers of the AIDS epidemic and were independently associated with age, race/ethnicity, and gender. Based on age seroprevalence trends, it was crudely estimated that at least one of 2,000 young men and one of 7,000 young women are infected with HIV annually in the U.S. Infection incidence rates, estimated from age and temporal trends, were estimated to be highest among black males (1.40/1,000/year) and lowest among white females (0.03/1,000/year). Poisson regression analysis of seroprevalence trends suggested that infection incidence rates accelerated among black females during the first 3 years of screening. Since selection factors undoubtedly changed over the period, estimates based on these data probably underestimate actual values in the general population, particularly near urban AIDS epicenters. Nonetheless, even crude estimates of these critical values, particularly among adolescents and young adults, are useful to guide policy development, to allocate resources, and to monitor program effects.

Evidence that adenosine contributes to maintenance of hepatosplanchnic blood flow during peritoneal sepsis in rats.

Sepsis induced derangements in hepatosplanchnic perfusion can contribute to organ damage and death. Adenosine, a common and potent metabolic vasodilator, has not been evaluated as a mechanism for maintenance of blood flow during sepsis. We tested the hypothesis that adenosine receptor blockade would cause a decrease in hepatosplanchnic blood flow during intraperitoneal (i.p.) sepsis in the rat. Rats (250-350 g) were catheterized for hemodynamic and blood flow measurements with tracer microspheres. Sepsis was induced with an i.p. injection of cecal material (150 mg/kg in D5W; 5 mL/kg), and baseline measurements were taken 24 h later. Animals then received either the adenosine receptor antagonist 8-PTH (10 mM, 1.5 mL/kg), its vehicle (1.5 mL/kg) or normal saline (1.5 mL/kg), intravenously, and measurements were repeated 1 and 10 min later. There was a significant increase in hepatosplanchnic portal resistance in septic animals given 8-PTH, with no change in mean arterial blood pressure (MAP) or heart rate. Regionally, there was a significant decrease in gastric, small intestinal, cecal, and pancreatic blood flow when compared with vehicle. Adenosine receptor blockade caused a significant reduction in hepatosplanchnic blood flow during sepsis, suggesting that maintenance of splanchnic blood flow during sepsis involves receptor mediated adenosine actions.

Laboratory evaluation of a dot-blot enzyme immunoassay for serologic confirmation of illness due to Rickettsia conorii.

Of the 169 United States Army soldiers who deployed on a field training exercise to a remote area of Botswana for two weeks in January 1992, more than 30% developed a febrile illness within five days of their return. A diagnosis of South African tick typhus was suggested by soldiers' exposure to ticks, as well as the presence of eschars and vesicles at the site of tick bites, and tender regional lymphadenopathies. This high attack rate, experienced during such a short exposure period, emphasizes the hazard of illness due to Rickettsia conorii to persons visiting endemic areas. A rapid, diagnostic, semiquantitative enzyme immunoassay (DS) for detection of IgG and IgM antibodies to R. conorii was performed on 209 acute and convalescent sera from soldiers in the outbreak and on 75 control sera. For the acute sera from soldiers meeting the probable case definition of having both regional lymphadenopathy and tick bite eschar, as judged by an IgG indirect fluorescent antibody (IFA) test, the resulting sensitivity and specificity of the DS test were 100% and 48%, respectively. In the analysis of the acute sera, the DS test identified as reactive more of the probable cases (62%) than either the IgG (16%) or IgM (55%) IFAs. This simple and rapid diagnostic test could be useful in establishing a preliminary diagnosis of R. conorii rickettsiosis in remote settings when immediate confirmation by IFA is impossible.

Rapid serodiagnosis of leptospirosis using the IgM-specific Dot-ELISA: comparison with the microscopic agglutination test.

The dot enzyme-linked immunosorbent assay (Dot-ELISA) was compared to the microscopic agglutination test (MA test) for the diagnosis of human leptospirosis. Of 177 sera from 68 soldiers who trained in the Republic of Panama, 102 sera were positive in the MA test and 93 of these sera were positive in the IgM-specific Dot-ELISA. Incidence of infection was 50 of 68 patients with the MA test and 48 of 68 in the IgM Dot-ELISA. Five MA test-positive sera were reactive only in the IgG-specific Dot-ELISA, suggesting previous exposure. All 21 infecting serovars of Leptospira interrogans, as determined by positive reactions in the MA test or culture of blood and urine specimens, were reactive in the Dot-ELISA. Of 75 sera negative in the MA test, 61 were nonreactive in the Dot-ELISA. However, 9 of these 14 Dot-ELISA-positive/MA test-negative sera were acute samples from patients whose later sera were MA test-positive. Positive reactions in the IgM Dot-ELISA occurred in 2 of 30 control, 4 of 10 Lyme disease, 1 of 11 relapsing fever, and 1 of 8 yaws sera; 10 syphilis patient sera were nonreactive. The IgM-specific Dot-ELISA appears to be sensitive and specific for the serodiagnosis of acute leptospirosis. In addition, this rapid test is inexpensive, simple to perform, utilizes minute volumes of killed leptospiral antigen and is easily adaptable to field use.

Epidemiologic investigation of an outbreak of cutaneous leishmaniasis in a defined geographic focus of transmission.

An outbreak of cutaneous leishmaniasis occurred in a unit of 608 Puerto Rican national guardsmen conducting jungle warfare training in the Panama Canal Area in July 1984. An epidemiologic investigation of reported nonhealing, ulcerating skin lesions was conducted among 540 (89%) unit members in November and December 1984. Fifteen (88%) of 17 individuals with chronic, ulcerating skin lesions were confirmed as cases of cutaneous leishmaniasis by culture or histopathology. Twelve cases yielded positive Leishmania cultures, identified as L. braziliensis panamensis by cellulose acetate electrophoresis. Evaluation of different diagnostic techniques revealed that direct examination of tissues by Giemsa-stained histological examination was the most sensitive test (87% sensitivity), with an indirect immunofluorescent antibody test being rather insensitive (67%). All but one of the confirmed cases operated in small units that trained and slept overnight at a mortar firing site for a period of three days, yielding a site-specific attack rate of 22% (14 of 64). This contrasted with a much lower attack rate of 0.2% (1 of 476), experienced by unit members who trained at other locations during the same time frame (P less than 0.001). The median incubation period calculated from day of arrival at the mortar firing site was 17 days (range 2-78) for the 15 confirmed cases. Available personal protection methods, such as the use of insect repellents, were not appropriately implemented by unit personnel and thus, were not found to effectively protect against Leishmania infection. This is the largest reported outbreak of cutaneous leishmaniasis in military personnel associated with a single geographic focus of infection and contrasts with the usual sporadic disease experience in Panama.

Concordance rates for benign prostatic disease among twins suggest hereditary influence.

OBJECTIVES: The etiology of benign prostatic hyperplasia (BPH) is unknown. Evidence for a hereditary trait would provide new avenues for investigation. METHODS: We compared the concordance for benign prostate disease in monozygotic (MZ) and dizygotic (DZ) twins who served in the United States military in World War II and have been followed by the Medical Follow-up Agency, Institute of Medicine of the National Academy of Sciences. Questionnaires completed in 1985 by 10,000 twins were reviewed for evidence of prostatic disease. Key words indicating benign prostatic disease (prostate, prostatectomy, BPH, TURP, and prostatism) were identified in 533 (5.3%) of the questionnaires. RESULTS: The average age was 64 +/- 3 years (range 56 to 68 in 1985). After eliminating men with known prostate cancer, there were 256 twin pairs that were informative for benign prostatic disease: both twins were concordant in 25 instances and discordant in 231, with only one twin mentioning benign prostatic disease. The pairwise concordance for MZ twins was 14.7% (19 of 129) and for DZ pairs it was 4.5% (5 of 112). The relative risk for benign prostatic disease for MZ twins was thus 3.3 (p = 0.008). The probandwise concordance rates, which express the probability of BPH in a cotwin of an affected twin, were 25.7% for MZ twins and only 8.5% for DZ twins. A covariance analysis estimated that 49% of the observed variance between twins could be attributed to genetic effects. CONCLUSIONS: These data provide preliminary evidence for the heritability of benign prostatic disease.

Acute and subacute effects of naturally occurring estrogens on luteinizing hormone secretion in the ovariectomized rat: Part 2.

Acute intravenous administration of the phytoestrogen genistein (G) blocks the gonadotropin-releasing hormone-(GnRH)-induced rise of luteinizing hormone (LH) in ovariectomized rats. The present experiments were performed to determine whether subacute administration of G or the mycoestrogens zearalenone and zearalenol would affect GnRH-induced or progesterone-induced LH secretion in ovariectomized rats. Charles River CD rats were ovariectomized and used 2 to 5 weeks later. Blood samples were obtained either via decapitation or via intraatrial cannulae three days after compounds were injected subcutaneously in sesame oil or corn oil vehicle. LH was measured by RIA. Pretreatment with estradiol benzoate suppressed LH levels at 1200 h, while G had no effect. Challenge with progesterone (8 mg/kg BW, sc) evoked LH release at 1600 h in rats pretreated with estradiol benzoate, but LH levels did not change in rats pretreated with G, zearalenone, or zearalenol. While GnRH-induced LH secretion was preserved in rats pretreated with estradiol, no LH response was detected in rats pretreated with the higher dose of G (8 mg/kg BW) or either dose of zearalenol (0.8 mg/kg BW or 8 mg/kg BW). We conclude that in the ovariectomized rat 1) subacute administration of G, zearalenone, or zearalenol do not inhibit tonic LH secretion, 2) G, zearalenone, and zearalenol do not provide "estrogenic priming" for progesterone-induced LH secretion; however, 3) G and zearalenol do block GnRH-induced LH secretion. The seemingly selective neuroendocrine effects of these naturally-occurring dietary estrogens emphasize that actions of each putative estrogen must be characterized for each "estrogenic" endpoint.

Observations on the biochemical effects of transfusion of citrate-phosphate-dextrose stored blood in man.

Preliminary data on the biochemical effects of the transfusion of three units of CPD blood in man is presented. Free (ionized) calcium was found to decrease approximately two minutes following initiation of the transfusion of CPD stored blood at a rate of 50 ml/min. Due to mixing problems, arterial and not venous specimens were found necessary to monitor accurately the severity of these decreases. Parathyroid hormone in the peripheral blood was found to increase two to four minutes after the initiation of transfusion. Decreases in blood pressure were observed during transfusion. The blood pressure recovered when the transfusion was stopped and did not decrease any further after restarting the transfusion, were found out not to increase or decrease consistently, and only modest changes in plasma potassium were observed.

Study on the effectiveness of remote mental healing.

A study which involved eight healers and ninety-six patients was conducted to determine the effectiveness of remote mental healing. The test subjects were hypertension patients between the ages of sixteen and sixty. Neither the doctor nor the patients knew who received the mental healing treatments. Normal medical treatment was continued in all cases. Improvement was judged by changes in the diastolic blood pressure, systolic blood pressure, heart beat rate, and weight. The statistical analysis showed a significant improvement in the systolic blood pressure of the healer-treated group compared with the change in the control group. There were no significant differences in the changes of diastolic blood pressure, pulse, and weight of the two groups. Four of the healers had a 92.3 per cent improvement ratio in their total group of patients compared with a 73.7 per cent improvement for the control group. The general healing treatment used by the healers involved (a) a relaxation step, (b) attunement with a Higher Power or Infinite Being, (c) a visualization and/or affirmation of the patient being in a state of perfect health, and (d) expression of thanks to God or to the Source of all power and energy.

Cirrhosis mortality among former American prisoners of war of World War II and the Korean conflict: results of a 50-year follow-up.

In our earlier, 30-year follow-up of American prisoners of war (POWs) of World War II and the Korean conflict, we found evidence of increased cirrhosis mortality. Using federal records, we have now extended our follow-up to 50 years (42 years for Korean conflict veterans) and have used proportional hazards analysis to compare the mortality experience of POWs with that of controls. Compared with their controls, World War II POWs had a 32% higher risk of cirrhosis mortality (statistically significant), and mortality risk was higher in the first 30 years of follow-up and also among those aged 51 years and older. Korean POWs had roughly the same risk of cirrhosis mortality as their controls. Neither self-reported data on alcohol consumption nor supplemental morbidity data satisfactorily explained the differences in risk between POWs and controls, although there was evidence that POWs tended to have higher rates of hepatitis, helminthiasis, and nutritional deprivation.

An outbreak of hookworm infection associated with military operations in Grenada.

During 1983, a multinational military intervention took place on Grenada. After deployment, troops from several U.S. Army units noted signs and symptoms consistent with soil-transmitted helminthic infection. Of 684 soldiers screened five to seven weeks post-deployment, over 20% reported abdominal pain and/or diarrhea during or after the action. Eosinophilia of at least 10% was observed in 119 (22.5%) of 529 soldiers evaluated further; eosinophilia of 5-9% was documented in another 126 (23.8%) of the 529 soldiers. Stool examinations confirmed hookworm infection in 35 soldiers. One case of strongyloidiasis was also documented. Infection was attributed to ground exposure near homes with compromised sanitation. Units that joined the operation after the initial assault phase were at low risk of hookworm infection.

The status of home care training for physician assistants.

Homebound individuals represent a medically underserved and rapidly growing population. Nonphysician practitioners--physician assistants and nurse practitioners--in the home care setting afford increased availability of and access to medical services by homebound patients.

Purification and characterization of a liver-derived beta-N-Acetylhexosaminidase from marine mammal Sotalia fluviatilis.

A beta-N-Acetylhexosaminidase (EC 3.2.1.52) was purified from hepatic extracts of Sotalia fluviatilis, order Cetacea. The protein was purified by using ammonium sulfate fractionation and four subsequent chromatographies (Biogel A 1.5 m, Chitin, Deae-Biogel and hydroxyapatite resins). After these purification steps, the enzyme was purified 380.5-fold with an 8.4% yield. The molecular mass (10 kDa) was estimated by SDS-PAGE and MALDI-TOF analysis. A Km of 2.72 mM and Vmax 9.5 x 10(-6) micromol/(min x mg) were found for this enzyme, determined by p-nitrophenyl-beta-D: -hexosaminide substrate digestion. Optimal pH and temperature for beta-N-Acetylhexosaminidase activity were 5.0 and 60 degrees C, respectively. Enzyme activity was inhibited by sodium selenate (Na(2)SeO(4)), mercuric chloride (HgCl(2)) and sodium dodecyl sulfate (C(12)H(25)SO(4)Na), and activated by zinc, calcium, barium and lithium ions. Characterization of the beta-N-Acetylhexosaminidase in Sotalia fluviatilis can be a basis for physiological studies in this species.