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PubMed1981

Developmental regulation of secretory protein synthesis in rat seminal vesicle.

This paper reports a developmental study of one of the major secretory proteins (SVS IV) of the rat seminal vesicle. Measured by radioimmunoassay, SVS IV is detectable even in very young rats (5 days old) but does not begin to accumulate in massive amounts until animals reach about 35 days of age. We have compared SVS IV synthesis by minced tissue from rats undergoing sexual maturation (25-60 days of age) with levels of translatable SVS IV mRNA present. Between 25 and 60 days of age, the proportion of newly synthesized protein devoted to SVS IV by tissue minces increased from 0.8% to 20%. In contrast, the proportion of total RNA devoted to translatable SVS IV mRNA was already high in the immature glands and increased less than 4-fold during development. These results seem to indicate that factors other than the presence of potentially translatable message regulate the rate of net synthesis of SVS IV during seminal vesicle maturation.

Aging↗
PubMed1983

Seminal vesicle secretion IV gene: allelic difference due to a series of 20-base-pair direct tandem repeats within an intron.

The rat seminal vesicle secretion IV (SVS IV) gene was isolated from a lambda Charon 4A library. The SVS IV gene transcription unit was found to be on one 3.3-kilobase (kb) EcoRI fragment. Restriction mapping and DNA sequence analysis demonstrated that the entire length of the SVS IV transcription unit is 1,930 base pairs (bp) and contains two introns. The 3.3-kb EcoRI fragment contains 144 bp of 5'-flanking region. At -113 bp from the presumed transcription initiation site an interesting structure with perfect dyad symmetry is noted. In another lambda clone, a 3.5-kb EcoRI fragment was isolated that contains the SVS IV gene and was shown to be identical to the 3.3-kb EcoRI fragment except for 180 bp of DNA in the second intron. The extra DNA consists of several (8-10) 20-bp tandem repeats flanked on each side by seven or eight copies of this same 20-bp repeat. Fisher X Sprague-Dawley hybrid rats, which contain both the EcoRI 3.5-kb form and the 3.3-kb form of the SVS IV gene, were crossed with each other. Analysis of the F1 generation demonstrated that the presence or absence of the 180-bp intronic insertion in the SVS IV gene defines an allelic difference. This report also presents the DNA sequence of the transcription unit and flanking regions of the SVS IV gene.

Alleles↗
PubMed2005

Exocytosis and endocytosis of synaptic vesicles and functional roles of vesicle pools: lessons from the Drosophila neuromuscular junction.

To maintain synaptic transmission during intense neuronal activities, the synaptic vesicle (SV) pool at release sites is effectively replenished by recruitment of SVs from the reserve pool and/or by endocytosis. The authors have studied dynamics of SVs using a fluorescence dye, FM1-43, which is incorporated into SVs during endocytosis and released by exocytosis. Drosophila is one of the most suitable preparations for genetic and pharmacological analyses, and this provides a useful model system. The authors found at the neuromuscular junctions of Drosophila that exocytosis and endocytosis of SVs are triggered by Ca(2+) influx through distinct routes and that selective inhibition of exocytosis or endocytosis resulted in depression of synaptic transmission with a distinct time course. They identified two SV pools in a single presynaptic bouton. The exo/endo cycling pool (ECP) is loaded with FM1-43 during low-frequency stimulation and locates close to release sites in the periphery of boutons, whereas the reserve pool (RP) is loaded and unloaded only during high-frequency stimulation and resides primarily in the center of boutons. The size of ECP closely correlates with the quantal content of evoked release, suggesting that SVs in the ECP are primarily involved in synaptic transmission. SVs in the RP are recruited to synaptic transmission by a process involving the cAMP/PKA cascade during high-frequency stimulation. Cytochalasin D blocked this recruitment process, suggesting involvement of filamentous actin. Endocytosed SVs replenish the ECP during stimulation and the RP after tetanic stimulation. Replenishment of the ECP depends on Ca(2+) influx from external solutions, and that of the RP is initiated by Ca(2+) release from internal stores. Thus, SV dynamics is closely involved in modulation of synaptic efficacy and influences synaptic plasticity.

Animals↗
PubMed2025

Structural variants linked to Alzheimer's disease and other common age-related clinical and neuropathologic traits.

BACKGROUND: Alzheimer's disease (AD) is a complex neurodegenerative disorder with substantial genetic influence. While genome-wide association studies (GWAS) have identified numerous risk loci for late-onset AD (LOAD), the functional mechanisms underlying most of these associations remain unresolved. Large genomic rearrangements, known as structural variants (SVs), represent a promising avenue for elucidating such mechanisms within some of these loci. METHODS: By leveraging data from two ongoing cohort studies of aging and dementia, the Religious Orders Study and Rush Memory and Aging Project (ROS/MAP), we performed genome-wide association analysis testing 20,205 common SVs from 1088 participants with whole genome sequencing (WGS) data. A range of Alzheimer's disease and other common age-related clinical and neuropathologic traits were examined. RESULTS: First, we mapped SVs across 81 AD risk loci and discovered 22 SVs in linkage disequilibrium (LD) with GWAS lead variants and directly associated with the phenotypes tested. The strongest association was a deletion of an Alu element in the 3'UTR of the TMEM106B gene, in high LD with the respective AD GWAS locus and associated with multiple AD and AD-related disorders (ADRD) phenotypes, including tangles density, TDP-43, and cognitive resilience. The deletion of this element was also linked to lower TMEM106B protein abundance. We also found a 22-kb deletion associated with depression in ROS/MAP and bearing similar association patterns as GWAS SNPs at the IQCK locus. In addition, we leveraged our catalog of SV-GWAS to replicate and characterize independent findings in SV-based GWAS for AD and five other neurodegenerative diseases. Among these findings, we highlight the replication of genome-wide significant SVs for progressive supranuclear palsy (PSP), including markers for the 17q21.31 MAPT locus inversion and a 1483-bp deletion at the CYP2A13 locus, along with other suggestive associations, such as a 994-bp duplication in the LMNTD1 locus, suggestively linked to AD and a 3958-bp deletion at the DOCK5 locus linked to Lewy body disease (LBD) (P = 3.36 × 10-4). CONCLUSIONS: While still limited in sample size, this study highlights the utility of including analysis of SVs for elucidating mechanisms underlying GWAS loci and provides a valuable resource for the characterization of the effects of SVs in neurodegenerative disease pathogenesis.

Humans↗
PubMed1990

Postnatal growth of mouse seminal vesicle is dependent on 5 alpha-dihydrotestosterone.

The seminal vesicles (SV) develop from the lower portion of the Wolffian ducts (WD) in response to androgens, which prevent their degeneration and subsequently stimulate organogenesis of the epididymis, vas deferens, seminal vesicles, and ejaculatory ducts. Earlier studies suggest that testosterone (T) is the active androgen for WD development. By contrast, development of urogenital sinus and external genitalia is dependent upon 5 alpha-dihydrotestosterone (DHT), produced by 5 alpha-reductase within the target tissue itself. To reevaluate the possible role of DHT during SV morphogenesis, SVs from 0-day-old (day of birth) mice were grown for 3, 6, or 9 days in either serum-free or serum-containing medium in the presence or absence of T (10(-7) M) or DHT (10(-8) M). The serum-free medium consisted of Ham's F-12-Dulbecco's Modified Eagle's Medium (1:1) containing insulin, transferrin, cholera toxin, BSA, and epidermal growth factor. The serum-containing medium was Ham's F-12-Dulbecco's Modified Eagle's Medium containing 10% fetal calf serum. Epithelial branching morphogenesis of SVs occurred in serum-free or serum-containing medium supplemented with either T or DHT and was comparable to that of SVs of similar ages in vivo. In serum-containing medium the DNA content of the cultures was about 2-fold higher in T-containing vs. T-deficient medium. However, in serum-free medium the DNA content was the same in cultures grown with or without T. SVs cultured under serum-free conditions in the presence of T plus 390 MSD (17 beta-N,N-diisopropylcarbamoyl-4-aza-5 alpha-androstan-3-one, an inhibitor of 5 alpha-reductase) were completely inhibited in their development, while in the presence of DHT plus 390 MSD, branching morphogenesis was comparable to that in SVs cultured in the presence of T or DHT alone. In medium lacking either T or DHT, SV development was inhibited. In addition, it was confirmed by TLC that [1 beta,2 beta-3H]T was converted into [3H]DHT at the ratio of 15.4% for the first 2 days and at 35.3% for the subsequent 2 days of the culture of SVs in serum-free medium. These data demonstrate that T is important as a precursor of DHT and DHT is the major androgen in the postnatal development of mouse SVs from the lower WD.

Animals↗
PubMed2007

The mouse seminal vesicle shape mutation is allelic with Fgfr2.

The mouse seminal vesicle shape (svs) mutation is a spontaneous recessive mutation that causes branching morphogenesis defects in the prostate gland and seminal vesicles. Unlike many other mutations that reduce prostatic and/or seminal vesicle branching, the svs mutation dramatically reduces branching without reducing organ growth. Using a positional cloning approach, we identified the svs mutant lesion as a 491 bp insertion in the tenth intron of Fgfr2 that results in changes in the pattern of Fgfr2 alternative splicing. An engineered null allele of Fgfr2 failed to complement the svs mutation proving that a partial loss of FGFR2(IIIb) isoforms causes svs phenotypes. Thus, the svs mutation represents a new type of adult viable Fgfr2 allele that can be used to elucidate receptor function during normal development and in the adult. In the developing seminal vesicles, sustained activation of ERK1/2 was associated with branching morphogenesis and this was absent in svs mutant seminal vesicles. This defect appears to be the immediate downstream effect of partial loss of FGFR2(IIIb) because activation of FGFR2(IIIb) by FGF10 rapidly induced ERK1/2 activation, and inhibition of ERK1/2 activation blocked seminal vesicle branching morphogenesis. Partial loss of FGFR2(IIIb) was also associated with down-regulation of several branching morphogenesis regulators including Shh, Ptch1, Gli1, Gli2, Bmp4, and Bmp7. Together with previous studies, these data suggest that peak levels of FGFR2(IIIb) signaling are required to induce branching and sustain ERK1/2 activation, whereas reduced levels support ductal outgrowth in the prostate gland and seminal vesicles.

Alleles↗
PubMed2006

Tooth bleaching by different concentrations of carbamide peroxide and hydrogen peroxide whitening strips: an in vitro study.

OBJECTIVE: To investigate the tooth whitening effects of various concentrations of carbamide peroxide (CP) gels and 6% hydrogen peroxide (HP) whitening strips used on an intrinsic, in vitro stain model in a simulated home-applied bleaching protocol. METHOD: Extracted third molars were sectioned and stained to Vita shade C4 using a standardized tea solution. Stained specimens were then bleached with 10, 15, 20, 22, and 30% CP gels applied in custom-made trays for 8-hour sessions for 14 days. A 6% HP whitening strip product was also tested in a regimen of twice-daily 30-minute treatments for 14 days. Shades were assessed at baseline and at 2, 5, 7, 10, and 14 days of treatment using a shade guide (SG) and a shade vision system (SVS), recorded as shade guide unit (SGU) changes from baseline, and CIE L*a*b* recordings using a chromometer. RESULTS: By day 14, all CP treatments resulted in at least 12 SGU improvements by SG and SVS methods: the HP treatment mean was just less than 12 SGU. With the chromometer, the CP improvements ranged from approximately 19 to 28 units and 16 units for the HP whitening strips. Observationally, by SG and SVS, CP treatments achieved the maximum improvement (12-13 SGU) at different time points: day 5 for 30% CP, day 10 for 22% CP, and day 14 for the other three treatments. SG and SVS data were virtually binary, switching from 0 to scores of 9 or above as bleaching progressed. The differences between the six treatments in the mean day to achieve a positive SG or SVS score (9 or more units) approached significance. For each of the SG, SVS, and L*a*b* scores, the dose-response correlation with CP concentration was significant at one or more assessment times. SG and SVS showed extremely strong agreement in detecting change and substantial agreement with L*a*b*. CONCLUSION: This in vitro study supports the limited data available from the very few available randomized controlled clinical trials indicating that CP and HP home-use bleaching systems can achieve considerable tooth whitening outcomes, albeit at different rates, which appear to be concentration dependent. CLINICAL SIGNIFICANCE: There is a clear significant relationship for both concentration and duration of exposure for CP bleaching agents. The final shade change is independent of the concentration of bleaching agent, with time as the dominant variable. Higher concentrations of CP that have not been investigated previously may be a treatment option for esthetic improvement of shade where time is at a premium, but caution must be exercised in view of the possible increased incidence of sensitivity.

Carbamide Peroxide↗
PubMed2026

RAG-mediated structural variation and its impact on relapse risk in acute lymphoblastic leukemia.

Relapse during treatment of B-cell acute lymphoblastic leukemia (B-ALL) is a harbinger of poor outcomes. Identifying biomarkers for subsequent relapse risk which are detectable at B-ALL diagnosis remains a priority. Off-target recombination-activating gene (RAG)-mediated structural variants (SVs) generate genomic instability that drives leukemogenesis and may underlie treatment resistance. Leveraging sequencing data in 1,496 pediatric B-ALL patients enriched for relapse status (relapse n=532; non-relapse n=964), we characterized RAG-mediated SVs across B-ALL molecular subtypes and examined their association with patient characteristics and their impact on clinical outcomes. Off-target RAG-mediated SVs were overall frequent, particularly in ETV6::RUNX1, ETV6::RUNX1-like, and Ph-like B-ALL subtypes, while increasing age-at-diagnosis was positively associated with burden of off-target RAG-mediated SVs (P<.001). Off-target RAG-mediated SVs with a recombination signal sequence (RSS) at one breakpoint, a hallmark of off-target RAG activity, were significantly more frequent at diagnosis in patients who subsequently relapsed (P=.001). This association remained significant in multivariable regression analysis (per SV odds ratio [OR]:1.08, 95%CI:1.04-1.12), in minimal residual disease (MRD)-negative patients (OR:1.09, 95%CI:1.04-1.14) and across subtypes. Excluding deletions, MRD-negative ETV6::RUNX1 patients with &#x2265;3 off-target RAG-mediated SVs had a >3-fold risk of relapse (hazard ratio:3.47, 95% CI:1.86-6.49). RAG-mediated SVs were also associated with relapse risk in T-cell ALL patients. Off-target RAG-mediated SV burden at diagnosis is a risk factor of relapse in pediatric ALL across molecular subtypes and independent of MRD status.

Journal Article↗
PubMed1999

Proton MR spectroscopy in patients with complex partial seizures: single-voxel spectroscopy versus chemical-shift imaging.

BACKGROUND AND PURPOSE: Proton MR spectroscopy has recently been applied to the evaluation of seizures, but few comparisons have been made between different clinical spectroscopic techniques. Our goal was to determine whether there is a significant difference between hippocampal NAA/(Cho+Cr) ratios obtained by single-voxel spectroscopy (SVS) and by chemical-shift imaging (CSI). METHODS: Twelve healthy adults and eight patients with complex partial seizures were studied on a 1.5-T MR scanner using a proton SVS method. Another 12 healthy adults and 10 patients with complex partial seizures were recruited for a proton CSI study, which was performed on a different 1.5-T MR system. The NAA/(Cho+Cr) ratio was calculated from the integral peak areas by curve fitting. The two-tailed t-test was used for statistical analysis. RESULTS: The mean value +/- standard deviation of the hippocampal NAA/(Cho+Cr) ratio in healthy control subjects was 0.63 +/- 0.07 by SVS, with 0.62 +/- 0.15 for the anterior hippocampus and 0.65 +/- 0.11 for the posterior hippocampus by CSI. There was no significant difference between the control group data obtained by SVS and those by CSI, nor was there a regional difference in the CSI NAA/(Cho+Cr) ratio in the hippocampus. Relative to the control group, the patients with seizures had a significant decrease in the NAA/(Cho+Cr) ratio in the abnormal hippocampus: -28% by SVS, and -24% in the anterior hippocampus and -18% in the posterior hippocampus by CSI. Proton SVS and CSI detected hippocampal abnormalities, unilateral or bilateral, in all patients of each group. CONCLUSION: Under similar measurement conditions, proton SVS and CSI provide similar NAA/(Cho+Cr) ratios among healthy control subjects, and they possess comparable ability for detecting hippocampal abnormalities in patients with complex partial seizures.

Adolescent↗
PubMed1996

Dynamics of synaptic vesicles in cultured spinal cord neurons in relationship to synaptogenesis.

The dynamics of synaptic vesicles (SVs) during the development of presynaptic specializations in cultured Xenopus spinal cord neurons was studied with the fluorescent vesicular probe FM1-43. In naive neurons that have not contacted synaptic targets, packets of SVs are distributed along the entire neurite and are quite mobile. The interaction with the synaptic target, such as a muscle cell or a latex bead coated with basic fibroblast growth factor, results in the localization and immobilization of SV packets at the contact site. Depolarization resulted in exocytosis of SVs in both naive and target-contacted neurites. Okadaic acid, a phosphatase inhibitor, caused a dispersal of SV packets in both naive and target-contacted neurites. Thus, prior to target contact, SVs are already organized into packets capable of release and recycling by a phosphorylation-dependent mechanism. Target interaction then recruits and anchors these functional SV packets into forming the presynaptic nerve terminal. With fluorescent phalloidin as a probe, F-actin was found to colocalize with SV clusters at bead-neurite contacts. Although okadaic acid caused a dispersal of SVs at the beads, F-actin localization there was relatively resistant to this drug treatment. This suggests that SVs become localized at the target by interacting with an actin-based cytoskeletal specialization in a phosphorylation-sensitive manner. The induction of this cytoskeletal specialization by the target may be an early event in presynaptic differentiation.

Actins↗
PubMed1985

Bioelectric potentials in the carotid body.

Simultaneous recordings of focal slow potentials (sVs) and chemosensory discharges were made from cat carotid body-nerve preparations in situ. Chemoreceptor stimulants (100% N2, asphyxia, NaCN, ACh and nicotine), and depressants (100% O2, spontaneous gasps and dopamine) changed receptor polarization. sVs evoked by stimulants had a negative polarity whereas depressants elicited positive deflections. There was a direct correlation between maximal frequency of chemosensory discharges and peak sV amplitude when NaCN injections or N2 inhalation were used. However, cholinergic agents, dopamine and substance P evoked sVs which lacked correlation in time-course, amplitude or polarity with changes in sensory frequency. After a 6-day carotid nerve crush, different stimuli still evoked sVs even in the absence of sensory discharges. Both sVs and chemosensory discharges were abolished after 1 h ischemia produced by ligature of carotid body blood vessels. Thus, sVs from carotid body chemoreceptors probably include a neuronal component (the generator potential) directly responsible for the origin of chemosensory discharges, and a non-neuronal component (receptor or secretory potentials) probably originating in glomus and/or sustentacular cells.

Acetylcholine↗
PubMed1994

Formation of synaptic vesicles.

Synaptic vesicles (SVs) are specialized secretory organelles used for the fast and focal signaling between nerve cells. They are small and homogeneous in size (50 nm), and contain non-peptide neurotransmitters such as glutamate, gamma-aminobutyric acid (GABA) and acetylcholine. The exocytosis of SVs occurs at low rates in resting nerve terminals and is greatly stimulated by depolarization-induced Ca2+ influx. Following exocytosis, SV membranes are rapidly retrieved, refilled locally with neurotransmitters and reused for the assembly of new SVs. Over the past few years, significant progress has been made in characterizing the molecular composition of SVs. From these studies, we know that SVs share a conserved set of membrane proteins with transport vesicles involved in other pathways. Furthermore, these findings have provided us with a new understanding about the evolutionary origin of SVs from recycling vesicles present in all cells.

Animals↗
PubMed2006

Single-vessel versus bifurcation stenting for the treatment of distal left main coronary artery disease in the drug-eluting stenting era. Clinical and angiographic insights into the Rapamycin-Eluting Stent Evaluated at Rotterdam Cardiology Hospital (RESEARCH) and Taxus-Stent Evaluated at Rotterdam Cardiology Hospital (T-SEARCH) registries.

BACKGROUND: Routine drug-eluting stent (DES) implantation has recently improved outcome in patients undergoing percutaneous treatment of left main (LM) coronary artery. However, even in the DES era, distal LM treatment remains an independent predictor of poor outcome. Whether single-vessel stenting (SVS) or bifurcation stenting (BS) should be performed to optimize treatment of such a lesion is unclear. METHODS: From April 2002 to June 2004, 94 patients affected by distal LM disease underwent percutaneous intervention at our institution either with SVS (n = 48) or BS (n = 46). The 2 groups were well balanced for all baseline characteristics but the extension of disease in the LM carina. RESULTS: After a median follow-up of 587 days (range, 328-1179), the cumulative incidence of MACE was similar between the 2 groups (31% in the BS vs 28% in SVS group, HR 0.96, 95% CI 0.46-1.49, P = .92), with no difference for the composite death/myocardial infarction or target vessel revascularization. After adjustment for confounders, the technique of stenting was not a predictor of either major adverse cardiac events or target vessel revascularization. Angiographic analysis--performed in 81% of eligible patients in SVS and 87% in the BS group--confirmed the equivalency between SVS versus BS. CONCLUSIONS: In consecutive patients undergoing catheter-based distal LM intervention, SVS or BS may perform equally under both clinical and angiographic perspective in current DES era.

Aged↗
PubMed2006

Reduction in deep vein reflux after concomitant subfascial endoscopic perforating vein surgery and superficial vein ablation in advanced primary chronic venous insufficiency.

BACKGROUND: Subfascial endoscopic perforating vein surgery (SEPS) and superficial vein surgery (SVS) have been the recommended treatment for advanced chronic venous insufficiency (CVI), despite a high prevalence of deep vein reflux in these patients. The anatomic and hemodynamic results of these procedures, however, remain uncertain. It is hypothesized that concomitant SEPS and SVS would result in a reduction of deep vein reflux in patients with advanced primary CVI. We investigated the effect of concomitant SEPS and SVS on deep vein reflux as well as the associated hemodynamic and clinical changes after surgery in a cohort of patients with advanced primary CVI. METHODS: We prospectively evaluated 53 consecutive SEPSs with concomitant SVS procedures in 47 patients with advanced primary CVI. There were 25 men and 22 women with a mean age of 58 years at operation. Thirty-four procedures (64%) were performed for limbs with active venous ulcers (class 6), and the other 19 procedures were performed for 15 class 5 limbs, one class 4a limb, and three class 4b limbs, respectively. Duplex scan and air plethysmography were performed before operation, at 1 month, and at 1 year after operation. The patients were followed up regularly with clinical assessment, and the ulcer healing and recurrence rates were documented. RESULTS: The proportion of limbs with common femoral vein incompetence decreased from 68% to 28% at 1 month and to 32% at 1 year after operation. The proportion of limbs with deep vein incompetence at more than one site also decreased from 42% to 15% at 1 month and to 12% at 1 year after concomitant SEPS and SVS. Venous hemodynamics as measured by air plethysmography improved significantly after operation. The cumulative ulcer healing was 85% at 3 months and 97% at 6 months. With a mean follow-up of 31 +/- 16 months, all ulcers healed. Only three recurrent ulcers (6%) were detected during the follow-up period. CONCLUSION: Concomitant SEPS and SVS are effective in reducing deep vein reflux and results in hemodynamic and clinical improvements in patients with advanced primary CVI. Deep vein reconstruction procedures may not be necessary in these patients.

Adult↗
PubMed2004

Mobilization and fusion of a non-recycling pool of synaptic vesicles under conditions of endocytic blockade.

At vertebrate central synapses, it has been demonstrated that a resting pool of synaptic vesicles (SVs) exists that normally does not participate in SV release and recycling. It remains unclear whether SVs within the resting pool are capable of mobilization and fusion. Here, we combine live imaging of SV exo- and endocytosis using pH-sensitive GFP (synapto-pHluorins) with pharmacological and genetic manipulations of the SV cycle at the Drosophila NMJ. We demonstrate that a resting pool of SVs exists at this synapse that encompasses 30-41% of the total SV pool. Under conditions of endocytic blockade, using a temperature-sensitive dynamin mutation, the resting pool of SVs can be mobilized and released. We present a model for the presence of a resting pool of SVs that does not require molecular specification of a subpopulation of SVs.

Animals↗
PubMed1994

Molecular feminization of mouse seminal vesicle by prenatal exposure to diethylstilbestrol: altered expression of messenger RNA.

Exposure to estrogens during critical stages of development has been reported to cause irreversible changes in estrogen target tissues such as the reproductive tract. In fact, recent studies using mice describe prenatal estrogen exposure resulting in the expression of the major estrogen-inducible uterine secretory protein, lactoferrin (LF), by the seminal vesicles of the male offspring. Thus, we have studied the role of estrogens in abnormal and normal gene expression in the developing male reproductive tract using LF and seminal vesicle secretory protein IV (SVS IV), an androgen-regulated murine seminal vesicle secretory protein, as markers. Lactoferrin and SVS IV protein and mRNA expression were studied in histological samples by using the techniques of in situ hybridization (ISH) and immunohistochemistry (IHC). Seminal vesicle secretory protein IV was expressed in all (100%) epithelial cells of the control seminal vesicle, but this protein was decreased by castration. However, LF expression was undetectable by ISH or IHC in control seminal vesicle epithelium. Lactoferrin was inducible in 2% of the seminal vesicle epithelial cells from adult castrated mice treated with estradiol 17 beta (E2; 20 micrograms/kg/day for 3 days), indicating that a small percentage of the seminal vesicle cells could be induced to secrete LF after modification of the endocrine environment. Prenatal DES treatment (100 micrograms./kg. maternal body weight on days 9 through 16 of gestation) resulted in the male offspring exhibiting constitutive expression of LF in 5% of the seminal vesicle epithelial cells, while expression of the androgen-regulated protein SVS IV was slightly decreased. The maximal contrast between LF and SVS IV expression was observed in prenatally DES-treated mice that were subsequently castrated as adults and further treated with E2; LF was detected in 40% of the epithelial cells in these mice. Double immunostaining techniques revealed that epithelial cells which were making LF had ceased production of SVS IV. Since a large percentage of the epithelial cells in the intact prenatal DES exposed male was capable of expressing the normal gene product, SVS IV, it was concluded that DES treatment during prenatal development appears to imprint or induce estrogenic sensitivity in the adult seminal vesicle, causing increased production of LF. The results suggest that this altered protein response may be an example of atypical gene expression in male reproductive tract tissues following hormonal manipulation early in development.

Animals↗
PubMed1994

Heparin and protamine use in peripheral vascular surgery: a comparison between surgeons of the Society for Vascular Surgery and the European Society for Vascular Surgery.

It was the intent of this study to document, in general, the patterns and complications of heparin and protamine usage during carotid endarterectomy, aortic and femoral-popliteal-tibial reconstructions for occlusive disease, elective and emergent abdominal aortic aneurysmectomy, thromboembolectomy, and dialysis arteriovenous (AV) fistula placement by surgeons from North America and Europe. All vascular surgeons from the Society for Vascular Surgery (SVS) and the European Society for Vascular Surgery (ESVS) were surveyed by a voluntary, self-reported questionnaire. Six hundred and forty-six completed questionnaires (284 from SVS and 362 from ESVS), representing a 62% response rate, were returned for evaluation. Systemic and regional administration of heparin was common during vascular procedures performed by both SVS and ESVS surgeons. Use of protamine to reverse heparin anticoagulation varied among SVS and ESVS surgeons, respectively, during: carotid endarterectomy (54% vs. 26%, p < 0.01), elective aortic reconstruction for occlusive disease (58% vs. 23%, p < 0.001), elective aortic reconstruction for abdominal aortic aneurysm (63% vs. 27%, p < 0.001), and femoral-popliteal-tibial reconstruction (44% vs. 15%, p < 0.001). Adverse reactions to protamine among the 25,219 and 12,902 cases reported from SVS and ESVS surgeons, respectively, included: hypotension (1209 and 495 cases), pulmonary artery hypertension (65 and eight cases), anaphylaxis (52 and 10 cases), and death (seven and two cases). These adverse responses accounted for 5.3% and 4.0% of the SVS and ESVS cases, respectively. Although this study is subject to the known limitations of a retrospective survey, it is clear that heparin use is common. Protamine reversal of heparin anticoagulation is more common in North America.(ABSTRACT TRUNCATED AT 250 WORDS)

Data Collection↗
PubMedDate not supplied

Two synaptic vesicle pools, vesicle recruitment and replenishment of pools at the Drosophila neuromuscular junction.

Drosophila neuromuscular junctions ( D NMJs) are malleable and its synaptic strength changes with activities. Mobilization and recruitment of synaptic vesicles (SVs), and replenishment of SV pools in the presynaptic terminal are involved in control of synaptic efficacy. We have studied dynamics of SVs using a fluorescent styryl dye, FM1-43, which is loaded into SVs during endocytosis and released during exocytosis, and identified two SV pools. The exo/endo cycling pool (ECP) is loaded with FM1-43 during low frequency nerve stimulation and releases FM1-43 during exocytosis induced by high K(+). The ECP locates close to release sites in the periphery of presynaptic boutons. The reserve pool (RP) is loaded and unloaded only during high frequency stimulation and resides primarily in the center of boutons. The size of ECP closely correlates with the efficacy of synaptic transmission during low frequency neuronal firing. An increase of cAMP facilitates SV movement from RP to ECP. Post-tetanic potentiation (PTP) correlates well with recruitment of SVs from RP. Neither PTP nor post-tetanic recruitment of SVs from RP occurs in memory mutants that have defects in the cAMP/PKA cascade. Cyotochalasin D slows mobilization of SVs from RP, suggesting involvement of actin filaments in SV movement. During repetitive nerve stimulation the ECP is replenished, while RP replenishment occurs after tetanic stimulation in the absence of external Ca(2+). Mobilization of internal Ca(2+) stores underlies RP replenishment. SV dynamics is involved in synaptic plasticity and D NMJs are suitable for further studies.

Animals↗