Effects of herpes simplex virus vectors encoding poreless TRPV1 or protein phosphatase 1α in a rat cystitis model induced by hydrogen peroxide.

Enhanced afferent excitability is considered to be an important pathophysiological basis of interstitial cystitis/bladder pain syndrome (IC/BPS). In addition, transient receptor potential vanilloid-1 (TRPV1) receptors are known to be involved in afferent sensitization. Animals with hydrogen peroxide (HP)-induced cystitis have been used as a model exhibiting pathologic characteristics of chronic inflammatory condition of the bladder. This study investigated the effect of gene therapy with replication-defective herpes simplex virus (HSV) vectors encoding poreless TRPV1 (PL) or protein phosphatase 1 α (PP1α), a negative regulator of TRPV1, using a HP-induced rat model of cystitis. HSV vectors encoding green fluorescent protein, PL or PP1α were inoculated into the bladder wall of female rats. After 1 week, 1% HP or normal saline was administered into the bladder, and the evaluations were performed 2 weeks after viral inoculation. In HP-induced cystitis rats, gene delivery of PL or PP1α decreased pain behavior as well as a reduction in the intercontraction interval. Also, both treatments reduced nerve growth factor expression in the bladder mucosa, reduced bladder inflammation characterized by infiltration of inflammatory cells and increased bladder weight. Taken together, HSV-mediated gene therapy targeting TRPV1 receptors could be effective for the treatment of IC/BPS.

Paired field and water measurements from drainage management practices in row-crop agriculture.

This paper describes a multi-site and multi-decadal dataset of artificially drained agricultural fields in seven Midwest states and North Carolina, USA. Thirty-nine research sites provided data on three conservation practices for cropland with subsurface tile drainage: saturated buffers, controlled drainage, and drainage water recycling. These practices utilize vegetation and/or infrastructure to minimize off-site nutrient losses and retain water in the landscape. A total of 219 variables are reported, including 90 field measurement variables and 129 management operations and metadata. Key measurements include subsurface drain flow (206 site-years), nitrate-N load (154 site-years) and other water quality metrics, as well as agronomic, soil, climate, farm management and metadata records. Data are published at the USDA National Agricultural Library Ag Data Commons repository and are also available through an interactive website at Iowa State University. These multi-disciplinary data have large reuse potential by the scientific community as well as for design of drainage systems and implementation in the US and globally.

Regulation of baboon fetal ovarian folliculogenesis by estrogen.

Although it is well established that formation of the pool of follicles available for ovarian function and fertility in adulthood in human and non human primates occurs in utero, our understanding of the regulation of fetal ovarian development is incomplete. Our laboratories have been instrumental in establishing the baboon as a model for the study of human reproductive endocrinology and showed that estrogen plays a central integrative role in regulating fetal-placental development. Therefore, we adapted our baboon model to study the role of estrogen on fetal ovarian development. Estrogen receptors alpha and beta were expressed in pregranulosa cells and interfollicular nests of the baboon fetal ovary. In baboons in which estrogen levels had been suppressed by administration of an aromatase inhibitor throughout the second half of gestation, fetal ovarian follicle numbers were reduced by 50%, whereas the number of interfollicular nests comprised of oocytes and pregranulosa cells was increased. The decrease in follicles in estrogen-deprived animals was associated with a marked upregulation of expression of alpha-inhibin, but not activins or activin receptors and signaling molecules. Moreover, the majority of the follicles formed in ovaries of estrogen-depleted fetuses appeared unhealthy and contained oocytes with a marked reduction/depletion in microvilli, structures essential for uptake of substrates from surrounding granulosa cells. We propose that estrogen regulates fetal ovarian folliculogenesis and formation of healthy oocytes by controlling the intraovarian activin:inhibin ratio and the development of oocyte microvilli. These findings demonstrate a need for translational research studies of the impact of impairment of estrogen action/availability on reproductive function in adulthood.

Isoform-specific mutations in the Caenorhabditis elegans heterochronic gene lin-14 affect stage-specific patterning.

The Caenorhabditis elegans heterochronic gene lin-14 specifies the temporal sequence of postembryonic developmental events. lin-14, which encodes differentially spliced LIN-14A and LIN-14B1/B2 protein isoforms, acts at distinct times during the first larval stage to specify first and second larval stage-specific cell lineages. Proposed models for the molecular basis of these two lin-14 gene activities have included the production of functionally distinct isoforms and the generation of a temporal gradient of LIN-14 protein. We report here that loss of the LIN-14B1/B2 isoforms alone affects one of the two lin-14 temporal patterning functions, the specification of second larval stage lineages. A temporal expression difference between LIN-14A and LIN-14B1/B2 is not responsible for the stage-specific phenotype: protein levels of all LIN-14 isoforms are high in early first larval stage animals and decrease during the first larval stage. However, LIN-14A can partially substitute for LIN-14B1/B2 when expressed at a higher-than-normal level in the late L1 stage. These data indicate that LIN-14B1/B2 isoforms do not provide a distinct function of the lin-14 locus in developmental timing but rather may contribute to an overall level of LIN-14 protein that is the critical determinant of temporal cell fate.

Role of RIHP and renal tubular sodium transporters in volume retention of pregnant rats.

BACKGROUND: Normal pregnancy is characterized by sodium and water conservation and an increase in plasma volume that is required for an uncomplicated pregnancy. Renal interstitial hydrostatic pressure (RIHP) is significantly decreased in pregnant rats. This decrease in RIHP may play an important role in the sodium and water retention that characterizes normal pregnancy. Paradoxically this enhanced renal sodium and water reabsorption appear to conflict with the consistent findings of a general decrease in abundance of renal tubular sodium transporters during normal pregnancy. The objective of this review is to examine the apparent discrepancy between the increases in renal tubular sodium and water reabsorption, facilitated by decreases in RIHP, and the seemingly discordant decreases in abundance of renal tubular transporters during normal pregnancy in rats. METHODS: Western blots and immunohistochemistry were used to evaluate abundance and localization of renal tubular transporters. RIHP was measured directly and continuously via a polyethylene (PE) matrix that was implanted in the left kidney of rats at the age of 11 to 16 weeks. RESULTS: Average basal RIHP and fractional excretion of sodium (FENa) were found to be significantly lower (P < .05) in midterm pregnant (MP; n = 18) and late-term pregnant (LP; n = 20) rats compared with nonpregnant (NP; n = 16) rats (3.5 +/- 0.3 mm Hg and 1.46 +/- 0.24% for MP; 3.3 +/- 0.1 mm Hg and 1.41 +/- 0.21% for LP; and 7.6 +/- 0.6 mm Hg and 3.67 +/- 0.24% for NP). Cortical Na+-K+-ATPase and Na-Pi2a cotransporter (Na-Pi) protein expression tend to decline with pregnancy. Also cortical Na+-H+ exchanger-1 (NHE-1) protein expression declines steadily during the course of pregnancy from MP to LP compared with that in NP rats, and cortical Na+-H+ exchanger-3 (NHE-3) protein expression is significantly lower in MP and LP compared with NP rats. CONCLUSIONS: We propose that during normal uncomplicated pregnancy, simultaneous decreases in RIHP and in net abundance of renal tubular sodium transporters occur. The effects of decreased RIHP exceed those of the reduction in net abundance, and presumably activity, of renal tubular transporters resulting in an enhanced net sodium and water retention during pregnancy.

Regulation of oocyte microvilli development in the baboon fetal ovary by estrogen.

We recently showed that the number of primordial follicles was reduced by 50% in ovaries of near-term fetal baboons deprived of estrogen in utero and restored to normal in animals supplemented with estrogen. Oocytes are avascular and rely on surrounding granulosa cells for nutrients, a process facilitated by microvilli on the oocyte surface. However, our understanding of oocyte microvillus development in the primate fetal ovary is incomplete. Thus, we determined whether estrogen regulates formation of oocyte microvilli in utero. Fetal ovaries were obtained on d 165 gestation (term = d 184) from baboons untreated (n = 3) or treated on d 100-165 with aromatase inhibitor CGS 20267 (estrogen suppressed by 95%; n = 5) or CGS 20267 and estradiol (n = 4). Follicles with intact (homogeneous cytoplasm) or nonintact (cytoplasm vacuolated) oocytes were quantified and the number/height of oocyte microvilli determined by electron microscopy. In untreated baboons, the mean (+/-se) number of follicles/0.08 mm(2) with an intact oocyte (11.5 +/- 0.5) was decreased (P < 0.05) by 70% in fetal ovaries of estrogen-suppressed baboons (3.4 +/- 0.2) and restored (P < 0.05) by CGS 20267 and estradiol (11.2 +/- 1.2). In estrogen-deprived fetuses, the number of microvilli/intact oocyte (23 +/- 3) was 56% lower (P < 0.01) than normal (52 +/- 5) and restored by CGS 20267 and estrogen (62 +/- 4). Moreover, in intact oocytes of estrogen-suppressed baboons, height (nm) of microvilli (105 +/- 11) was 54-62% lower (P < 0.01) than in intact oocytes of fetal ovaries of untreated (228 +/- 13) or estrogen-treated (274 +/- 17) baboons. In estrogen-replete baboons, the number of microvilli in intact oocytes was 2-fold greater (P < 0.01) than in nonintact oocytes. However, in estrogen-deprived baboons, no microvilli were detected in nonintact oocytes and the number of microvilli in intact oocytes was similar to that in nonintact oocytes of untreated fetuses. We conclude that development of microvilli in oocytes of primordial follicles in the primate fetal ovary is regulated by estrogen. Collectively, these results and those of our previous studies indicate that estrogen regulates fetal ovarian folliculogenesis and development of follicles with oocytes composed of microvilli critical for nutrient uptake and presumably long-term survival.

CFTR mRNA and its truncated splice variant (TRN-CFTR) are differentially expressed during collecting duct ontogeny.

The collecting duct epithelium originates from the embryonic ureter by branching morphogenesis. Ontogeny-dependent changes of CFTR mRNA expression were assessed by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in primary monolayer cultures of rat ureteric buds (UB) and cortical collecting ducts, microdissected at different embryonic and postnatal developmental stages. The amount of wild-type CFTR-specific PCR product in UB declined to 20% of the initial value between embryonic gestational day E15 and postnatal day P1. After birth the CFTR product increased transiently between P1 and P7 by a factor of 10 and decreased towards day P14. PCR products specific for TRN-CFTR, a truncated splice variant, however, were low in early embryonic cells, increased markedly between day E17 and P2, and reached a plateau postnatally. Therefore, mRNA encoding TRN-CFTR does not appear to have a specific embryonic-morphogenetic function. By contrast, such function is suggested for wild-type CFTR mRNA as its abundance was high in early embryonic nephrogenesis, as well as during a postnatal period shortly before branching morphogenesis is completed.

Neuropsychologic correlates of MRI white matter hyperintensities: a study of 150 normal volunteers.

To determine the effects of MRI white matter hyperintensities (WMH) on cognitive functioning, we used neuropsychologic tests and MRI to study 150 elderly volunteers free of neuropsychiatric or general disease. There were 76 (50.3%) individuals without and 74 (49.7%) with WMH. The latter subset was older (61.3 +/- 6.6 years versus 58.5 +/- 5.8 years, p = 0.005), had a higher mean arterial blood pressure (103.7 +/- 11.4 mm Hg versus 99.9 +/- 10.3 mm Hg, p = 0.03), and a larger ventricular-to-intracranial-cavity ratio (6.3 +/- 5.6% versus 4.7 +/- 1.6%, p = 0.02). Individuals with WMH performed worse than their counterparts without such abnormalities on all tests administered. After adjusting for the group differences in age, arterial blood pressure, and ventricular size, we noted statistically significant results on form B of the Trail Making Test (121.8 +/- 37.8 msec versus 100.3 +/- 47.9 msec, p = 0.04), a complex reaction time task (680.8 +/- 104.9 msec versus 607.1 +/- 93.9 msec, p = 0.001), and the assembly procedure of the Purdue Peg-board Test (27.5 +/- 5.8 versus 30.6 +/- 5.9, p = 0.02). Partial correlations did not reveal any relationship between test scores and the semiautomatically assessed total area of WMH. Our data suggest that the presence of WMH exerts a subtle effect on neuropsychologic performance of normal elderly individuals, which becomes particularly evident on tasks measuring the speed of more complex mental processing.

The Mattis Dementia Rating Scale: normative data from 1,001 healthy volunteers.

We administered the Mattis Dementia Rating Scale (MDRS) to 1,001 healthy volunteers, aged 50 to 80 years, randomly selected from our community. Multivariate regression analysis revealed educational level (p = 0.000004) and age (p = 0.00001), but no other sociodemographic or risk factors for stroke, to be significantly associated with the MDRS score. The age- and education-specific lowest quintile cutoff scores ranged from 140 in subjects aged 50 to 59 years with at least college experience to 130 in subjects aged 70 to 80 years with only 4 to 9 years of schooling. These percentile distributions obtained for decades of age and different levels of education should be useful reference values for clinicians and investigators when applying the MDRS to assess cognitive functioning.

Mini mental state examination: influence of sociodemographic, environmental and behavioral factors and vascular risk factors.

Age and education have been found to affect the Mini Mental State Examination (MMSE) score of elderly normals, but there have been no studies assessing the influence of environmental and behavioral factors on this test. We therefore administered the MMSE to 1437 normal elderly subjects in the setting of a stroke prevention study and correlated their results to 16 sociodemographic, environmental, and behavioral factors, and vascular risk factors. Study statistics composed of a multiple logistic regression analysis and graphical models revealed the relations between variables in greater detail. Logistic regression yielded education level, occupational status, living as a single, general life stress, physical strain, and physical inactivity to be independent predictors of the MMSE score. Age was not included in this model. Graphical models demonstrated similar results, but did not include living as a single and physical inactivity. As shown in our independence graph, general life stress is the crucial predictor and links other environmental and sociodemographic variables with the test performance of elderly normals.

Estrogen replacement therapy in older women: a neuropsychological and brain MRI study.

OBJECTIVE: To determine if postmenopausal women receiving estrogen perform better on demanding cognitive tests than women without estrogen replacement and if this beneficial effect on cognition is caused by estrogen-related prevention of silent ischemic brain damage. DESIGN: Cross-sectional study comparing postmenopausal estrogen users and non-users. SETTING: Austrian Stroke Prevention Study. PARTICIPANTS: A total of 70 women currently using estrogen and 140 women who have never used estrogen from a subset of 222 postmenopausal women without neuropsychiatric or general disease undergoing extensive diagnostic work-up in a large-scale stroke prevention study among randomly selected community members. MEASUREMENTS: Neuropsychological test scores and focal brain abnormalities as well as size of ventricles and cortical sulci as assessed by 1.5 Tesla MRI. RESULTS: Estrogen users performed better than non-users on almost all neuropsychological tests administered. When ANCOVA was used to correct for slight differences between groups in age, length of education, mean arterial blood pressure and self-reported activation, values of P < .05 were noted on tasks assessing conceptualization, attention, and visuopractical skills. After adjustment for multiple comparisons, the differences in conceptualization and visuopractical skills remained significant. MRI showed a lower rate and extent of white matter hyperintensities and a significantly smaller total white matter hyperintensity area in women treated with estrogen (P = .043). The total white matter hyperintensity area was inversely related to the duration of estrogen replacement therapy(P = .040). However, there was no difference in neuropsychological performance between estrogen users with and without white matter abnormalities, and this was also supported by the lack of an association between cognitive test results and the extent of white matter disease. CONCLUSIONS: Our study demonstrated an association between estrogen replacement therapy and better cognitive functioning and a lower rate of clinically unsuspected ischemic brain damage in postmenopausal women.

Plasma antioxidants and cognitive performance in middle-aged and older adults: results of the Austrian Stroke Prevention Study.

OBJECTIVES: To study the association between cognitive status and plasma concentrations of various antioxidants in middle-aged and older individuals without neuropsychiatric disease. DESIGN: Evaluation of cross-sectional data from a cohort study. SETTING: The Austrian Stroke Prevention Study. PARTICIPANTS: A total of 1769 subjects aged 50 to 75 years, with no history or signs of neuropsychiatric disease, selected randomly from the community register. MEASUREMENTS: The score on the Mattis Dementia Rating Scale (MDRS) was dichotomized according to age-and education-specific lowest quartile cut-off points. Reversed-phase high performance liquid chromatography measurements of the plasma concentrations of lutein/zeaxanthin, cryptoxanthin, canthaxanthin, lycopene, alpha-carotene, beta-carotene, retinol, gamma-tocopherol, alpha-tocopherol, and ascorbate were measured. RESULTS: Individuals with MDRS results below the lowest quartile cut-off point had lower levels of beta-carotene and alpha-tocopherol than their counterparts with test performance above this limit (0.44+/-.33 micromol/L vs 0.51+/-.48 micromol/L, P < .001; and 29.50+/-7.98 micromol/L vs 30.93+/-11.10 micromol/L, P < .001, respectively). Only alphatocopherol remained significantly associated with cognitive functioning when logistic regression analysis was used to adjust for possible confounders including age, sex, month of blood sampling, years of education, smoking, lipid status, and major risk factors for stroke (P = .019). CONCLUSION: These observations are compatible with the view that some dietary antioxidants may protect against cognitive impairment in older people.

The impact of sociodemographic, environmental, and behavioral factors, and cerebrovascular risk factors as potential predictors of the mattis dementia rating scale.

BACKGROUND: Age and education have been found to affect the Mattis Dementia Rating Scale (MDRS) score of elderly normals, but there have been no studies assessing the influence of environmental and behavioral factors on this scale. Their role as potential predictors of the MDRS total score was investigated. METHODS: The MDRS was administered to 1,927 normal elderly subjects in the setting of a stroke prevention study. Results were correlated with 16 sociodemographic, environmental, and behavioral factors, and cerebrovascular risk factors. Study statistics resulted from multiple logistic regression analysis. RESULTS: Results indicated that higher age and arterial hypertension were associated with poorer cognitive performance, while better education and moderate general life stress exerted a positive effect on the participants' test results. CONCLUSIONS: Thus, besides the well-established factors of age and educational level, moderate general life stress and hypertension were identified as relevant predictors in determining the MDRS test performance of elderly normals.

[Clinical diagnosis of normal pressure hydrocephalus (author's transl)]. / Klinik und Diagnose des Normaldruckhydrozephalus.

5 patients are described in which normal pressure hydrocephalus was diagnosed. The criteria on which the diagnosis was based were a cerebrospinal fluid perfusion test and an air study. Pathological verification was additionally obtained in 2 patients. The clinical picture in all patients was characterized by dementia. 4 patients were incontinent and 3 were paraspastic.

Cerebrovascular risk factors in an elderly Austrian population: first year results of the Austrian Stroke Prevention Study (ASPS).

During the first year the Austrian Stroke Prevention Study enrolled 599 volunteers without clinical signs or symptoms of cerebrovascular disease aged 50 to 70 years. Study participants were randomly selected from the official register of the city of Graz. The rate of positive response was 26.9 percent. All subjects underwent an extensive risk factor screening with Duplex scanning of the carotid arteries obtained from a subset of 176 individuals. The prevalence of well-documented cerebrovascular risk factors was 40.6% for arterial hypertension, 35.4% for cardiac disease, 8.5% for diabetes mellitus und 3% for elevated haematocrit. The less well-documented cerebrovascular risk factors dyslipidemia, overweight, physical inactivity, hyperfibrinogenemia and smoking were noted in 75%, 33.7%, 27.2%, 14.9% and 12.2% of subjects, respectively. Multiple well-documented risk factors were noted in 23.7% of the examined volunteers. Multiple linear regression analysis revealed body mass index (p < 0.0001) and age (p < 0.0001) as independent predictors of the frequency of well-documented risk factors observed in any individual. Atherosclerotic carotid disease occurred in 61.9% of study participants investigated by Doppler sonography and was significantly associated with age (p < 0.00001), life-time tobacco consumption (p < 0.0001) and the concentration of apolipoprotein B (p < 0.05). This study demonstrates high prevalence rates of vascular risk factors in an elderly Austrian community. Implications for stroke prevention result from the conjunction of overweight and frequency of risk factors noted in any study participant, as well as from the relationship of carotid atherosclerosis to smoking and dyslipidemia.

Evaluation of artificial insemination techniques on fertility in laying hens.

The effects of depth of intravaginal insemination of two subsequent semen ejaculates from Rhode Island Red (RIR) and Light Sussex (LS) cockerels on fertility and embryonic mortality of eggs from RIR hens were investigated. Intravaginal insemination to a depth of 3.5 cm resulted in significantly higher fertility than shallow insemination to .5 cm (74.0 vs. 62.3%). Although not statistically significant, fertilizing capacity of first ejaculate of semen collected from either RIR or LS males was greater than the second ejaculate (71.4 vs. 64.9%).

Poult yolk sac amount and composition upon placement: effect of breeder age, egg weight, sex, and subsequent change with feeding or fasting.

Hatching eggs from 48 and 55-week-old Small White turkey breeders were used to examine the yolk sac of these poults. Influences due to flock age, egg weight, poult sex, and a two day postplacement regimen of feeding or fasting were evaluated in a factorially designed experiment. Each variable was found to act independent of the other. Older breeders gave eggs having a greater percentage yolk and with more ether extract than if they were from younger hens. For the most part, these fundamental differences remained with the poult's yolk sac at and after placement. Examining egg weight effects apart from hen age showed that heavier eggs had heavier yolks but their percentage of the total was to the converse and no compositional differences appeared. Poults from large eggs had a greater amount of yolk sac that contained proportionately less ether extract than if they were from small eggs. In total, yolk sac found ca. 12 h after poult removal from the hatcher corresponded to reserves that would remain ca. two days after formal hatching. The yolk sac free carcass failed to exhibit any compositional differences regardless of egg source or weight upon placement, but 48 hr later changes occurred that could largely be attributed to interim yolk resorption. Fasting delayed yolk sac uptake and favored removal of moisture and lipid to a greater extent than protein while the converse was true if access to feed and water was permitted. All parameters were unaltered because of poult sex.

Hatching performance of Cryovac enclosed hatching eggs stored in a high humidity environment.

Two trials were performed to assess hatching success of enclosing White Leghorn hatching eggs in Cryovac for 3 to 4 weeks in a storage environment in excess of 90% relative humidity. Three treatments in each trial were carried out: trial 1, no Cryovac and Cryovac packing 1 and 4 days after lay; trial 2, Cryovac packing 1, 4, and 7 days after lay. No significant differences were detected for hatching performance between either non-Cryovac packed eggs or the time they were Cryovac packed after lay. As storage time increased from 22 to 28 days, the average hatch of fertile eggs for all treatments in both trials declined linearly (P less than .01) from 81.7 to 71.0%. This decline was mainly due to the linear increase (P less than .01) in early embryo mortality (1 to 7 days) from 11.0 to 21.0%.

The effect of caproic acid, handling technique and storage times on the fertility of fowl spermatozoa.

The effects of three levels of caproic acid in glycine-citrate extender on preservation of fertilizing capacity of fowl spermatozoa were tested. Extended semen stored for 6 and 28 hours was used for insemination in diluted or reconcentrated forms. The results indicated that semen stored in the extender with the highest concentration of caproic acid provided superior fertility and showed the importance of proper handling of hens during insemination.

The lethal nature of "wiry" mutation in the domestic fowl.

A study of the effect of normal environmental conditions on newly hatched chicks with the feather mutant described as "wiry" (Fiser et al., 1973) was carried out. The results indicated that the abnormality should be classified as a potential facultative lethal expressed after hatching.