Noninvasive experimental bladder pain assessment in painful bladder syndrome.

OBJECTIVE: To compare bladder sensitivity between patients with pelvic pain and patients who were pain free, undergoing noninvasive, controlled bladder distension via diuresis. We also sought to measure potential mechanisms underlying bladder sensitivity. DESIGN: Prospective observational study. SETTING: Community teaching hospital. POPULATION: Reproductive-age women with non-bladder chronic pelvic pain (CPP, n = 23), painful bladder syndrome (PBS, n = 23), and pelvic pain-free controls (n = 42) METHODS: Participants were compared on cystometric capacity, pelvic floor pressure-pain thresholds (PPTs), pelvic muscle function, O'Leary-Sant bladder questionnaire, and psychosocial instruments using Wilcoxon rank-sum tests. Multivariate regression was used to identify factors underlying bladder pain phenotypes. MAIN OUTCOME MEASURES: Pelvic floor pain thresholds; self-reported bladder distension pain. RESULTS: Participants with PBS exhibited higher bladder distension pain than those with CPP, with both groups reporting higher pain levels than controls (P < 0.05). No significant associations were found between bladder distension pain and pelvic muscle structure or pain sensitivity measures; however, bladder distension pain positively correlates with both vaginal PPTs adjacent to the bladder (r = 0.46) and pain with transvaginal bladder palpation (r = 0.56). Pain at maximal distension was less influenced by somatic sensitivity than bladder symptoms (r = 0.35 versus r = 0.59; P < 0.05). Multivariate regression identified three independent components of bladder symptoms in PBS: bladder distension pain, bladder sensation, and somatic symptoms. CONCLUSIONS: Diuresis-induced bladder pain differentiates CPP from PBS. Experimental bladder pain is not predicted by pelvic floor sensitivity. Compared with patient-reported outcomes it appears less influenced by psychological factors. Further study is needed to determine whether screening for experimental bladder pain sensitivity could predict future risk of PBS. TWEETABLE ABSTRACT: Controlled, water ingestion-provoked bladder pain can objectively identify visceral pain sensitivity.

Dilute versus concentrated vasopressin administration during laparoscopic myomectomy: a randomised controlled trial.

OBJECTIVE: To determine if higher-volume, fixed-dose administration of vasopressin further reduces blood loss at the time of minimally invasive myomectomy. DESIGN: Randomised multicentre clinical trial. SETTING: Tertiary-care academic centres in the USA. POPULATION: Women undergoing conventional laparoscopic or robot-assisted laparoscopic myomectomy. METHODS: All participants received the same 10-unit (U) dose of vasopressin, but were randomly assigned to one of two groups: (i) received 200 ml of diluted vasopressin solution (20 U in 400 ml normal saline), and (ii) received 30 ml of concentrated vasopressin solution (20 U in 60 ml normal saline). MAIN OUTCOME MEASURES: The primary study outcome was estimated blood loss; the study was powered to detect a 100-ml difference. RESULTS: A total of 152 women were randomised; 76 patients in each group. Baseline demographics were similar between groups. The primary outcome of intraoperative blood loss was not significantly different, as measured by three parameters: surgeon estimate (mean estimated blood loss 178 ± 265 ml and 198 ± 232 ml, dilute and concentrated groups respectively, P = 0.65), suction canister-calculated blood loss, or change in haematocrit levels. There were no vasopressin-related adverse events. CONCLUSION: Both dilute and concentrated vasopressin solutions that use the same drug dosing demonstrate comparable safety and tolerability when administered for minimally invasive myomectomy; however, higher volume administration of vasopressin does not reduce blood loss. TWEETABLE ABSTRACT: This randomised trial failed to show benefit of high-volume dilute vasopression.

On the magnetic properties of iron nanostructures fabricated via focused electron beam induced deposition and autocatalytic growth processes.

We employ Electron beam induced deposition (EBID) in combination with autocatalytic growth (AG) processes to fabricate magnetic nanostructures with controllable shapes and thicknesses. Following this route, different Fe deposits were prepared on silicon nitride membranes under ultra-high vacuum conditions and studied by scanning electron microscopy (SEM) and scanning transmission x-ray microspectroscopy (STXM). The originally deposited Fe nanostructures are composed of pure iron, especially when fabricated via autocatalytic growth processes. Quantitative near-edge x-ray absorption fine structure (NEXAFS) spectroscopy was employed to derive information on the thickness dependent composition. X-ray magnetic circular dichroism (XMCD) in STXM was used to derive the magnetic properties of the EBID prepared structures. STXM and XMCD analysis evinces the existence of a thin iron oxide layer at the deposit-vacuum interface, which is formed during exposure to ambient conditions. We were able to extract magnetic hysteresis loops for individual deposits from XMCD micrographs with varying external magnetic field. Within the investigated thickness range (2-16 nm), the magnetic coercivity, as evaluated from the width of the hysteresis loops, increases with deposit thickness and reaches a maximum value of ∼160 Oe at around 10 nm. In summary, we present a viable technique to fabricate ferromagnetic nanostructures in a controllable way and gain detailed insight into their chemical and magnetic properties.

Characterization and evolution of the mitochondrial DNA control region in Ranidae and their phylogenetic relationship.

The control region is considered to be one of the most variable parts of animal mitochondrial DNA (mtDNA). We compared the mtDNA control region from 37 species representing 14 genera and 4 subfamilies of Ranidae, to analyze the evolution of the control region and to determine their phylogenetic relationship. All the Ranidae species had a single control region, except four species that had two repeat regions. The control region spanned the region between the Cyt b and tRNAleu genes in most of the Ranidae species. The length of the control region sequences ranged from 1186 bp (Limnonectes bannaensis) to 6746 bp (Rana kunyuensis). The average genetic distances among the species varied from 1.94% (between R. chosenica and R. plancyi) to 113.25% (between Amolops ricketti and Euphlyctis hexadactylus). The alignment of three conserved sequence blocks was identified. However, conserved sequence boxes F to A were not found in Ranidae. A maximum likelihood method was used to reconstruct the phylogenetic relationship based on a general time reversible + gamma distribution model. The amount of A+T was higher than G+C across the whole control region. The phylogenetic tree grouped members of the respective subfamilies into separate clades, with the exception of Raninae. Our analysis supported that some genera, including Rana and Amolops, may be polyphyletic. Control region sequence is an effective molecular mark for Ranidae phylogenetic inference.

Mitogenome of Fejervarya multistriata: a novel gene arrangement and its evolutionary implications.

In this study, we determined the complete nucleotide sequence of the mitochondrial (mt) DNA of the paddy frog Fejervarya multistriata. mtDNA is 17,750-bp long and contains 13 protein-coding regions, 2 ribosomal RNA, non-coding genes, and 23 tRNA because of the presence of an extra copy of tRNA-Met. The gene arrangements among two related species of Fejervarya were compared, and the combined mtDNA data were subjected to a phylogenetic analysis. Interestingly, we observed a unique translocation of the tRNA-Leu gene, similar to that reported in previous studies on two Fejervarya species. Phylogenetic analyses supported the classification into two evolutionary clades, Ranidae and Dicroglossidae, as well as placement of Hylarana guentheri in the genus Babina. Our results suggested that Fejervarya limnocharis and Fejervarya multistriata may be conspecific, because of its low pairwise genetic distance. However, these results must be further validated with additional analyses.

DNA barcoding and phylogenetic relationships of genera Picoides and Dendrocopos (Aves: Picidae).

Picoides and Dendrocopos are two closely related genera of woodpeckers (family Picidae), and members of these genera have long been the subjects of phylogenetic debate. Mitochondrial cytochrome c oxidase subunit I (COI) is a powerful marker for the identification and phylogenetic study of animal species. In the present study, we analyzed the COI barcodes of 21 species from the two genera, and 222 variable sites were identified. Kimura two-parameter distances were calculated between barcodes. The average interspecific genetic distance was more than 20 times higher than the average intraspecific genetic distance. The neighbor-joining method was used to construct a phylogenetic tree, and all of the species could be discriminated by their distinct clades. Picoides arcticus was the first to split from the lineage, and the other species were grouped into two divergent clades. The results of this study indicated that the COI genetic data did not support the monophyly of Picoides and Dendrocopos.

miR-34a targets the inhibin beta B gene, promoting granulosa cell apoptosis in the porcine ovary.

During ovarian follicular growth and development, only a few follicles actually ovulate. Recently, it was found that follicular atresia is triggered by granulosa cell apoptosis, but the molecular mechanism of follicular atresia was not understood. Using flow cytometry, we found that miR-34a promotes granulosa cell apoptosis in pig ovarian follicles. In addition, inhibin beta B was found to be a miR-34a target gene, based on luciferase reporter assays, quantitative RT-PCR and Western blotting. Taken together, our data indicate that miR-34a plays an important role in granulosa cell apoptosis by targeting the INHBB gene in the porcine ovary.

Pharmacokinetics of intravenous and topical lidocaine in patients undergoing thoracoscopic pulmonary resection: a comparative study.

OBJECTIVE: Lidocaine was the commonly used local anesthetic. The present study aimed to compare the pharmacokinetics of intravenous and topical lidocaine in patients undergoing thoracoscopic pulmonary resection. PATIENTS AND METHODS: Sixty patients who were scheduled for thoracoscopic pulmonary resection were screened and randomly assigned to the intravenous lidocaine group and topical lidocaine group. After induction, the patient in the intravenous group was given an intravenous bolus of 1.5 mg/kg lidocaine, while the patient in the topical group was given 3.0 mg/kg lidocaine via the "spray-as-you-go" method. Arterial blood was sampled at preset intervals, and plasma concentrations of lidocaine and its metabolites [monoethylglycinexylidide (MEGX) and glycinexylidide (GX)] were measured by ultra-performance liquid chromatography-tandem mass spectrometry. RESULTS: Following intravenous administration, plasma lidocaine concentration reached its peak with a time to reach Cmax (Tmax) of 0.05 h and then decreased in a biphasic manner with a very short half-life time (T1/2) of 1.85 h. After topical administration, lidocaine was well absorbed, with Tmax of 0.21 h and bioavailability of 71.02%. The mean Tmax, Cmax, and area under the curve from the time (AUC0-t) of MEGX and GX were higher in the topical group than in the intravenous group. There were no obvious differences in the Cmax, T1/2, clearance, or apparent volume of distribution of lidocaine between the two groups. No obvious adverse events were observed. CONCLUSIONS: Topical administration of 3 mg/kg lidocaine via the "spray-as-you-go" method is an effective and safe technology for patients undergoing thoracoscopic pulmonary resection.

Effect of oral immunization with Aeromonas hydrophila ghosts on protection against experimental fish infection.

AIMS: To investigate whether oral immunization with Aeromonas hydrophila ghosts (AHG) vaccine can elicit mucosal and systemic immune responses of Carp (Carassius auratus gibelio) compared to conventional formalin-killed bacteria (FKC). METHODS AND RESULTS: Fish were fed diets coated with AHG, FKC or phosphate buffered saline (PBS) alone, after immunization, more antigen-specific antibody was significantly detected in serum and intestinal mucus in AHG group than FKC group and PBS group. In addition, after challenged with the parent strain J-1, the survival of bacterial ghost-vaccinated fish was higher than PBS group and FKC group, the relative per cent survival (RPS) being 76.8%, 58.9%, respectively. CONCLUSIONS: Oral immunization with A. hydrophila ghosts can elicit systemic and mucosal adaptive immune responses and has higher potential to induce protective adaptive immunity than normal vaccine. SIGNIFICANCE AND IMPACT OF THE STUDY: Oral immunization with bacterial ghosts is a promising new solution with potential application to prevent diseases in fish.

Evidence for clonal deletion and clonal anergy after intrathymic antigen injection in a transplantation model.

Intrathymic (IT) antigen injection has been shown to induce antigen-specific systemic tolerance in the rodent. To delineate the mechanisms responsible for the induction of tolerance, we used the 2C line of T cell receptor transgenic mice. The majority of T cells in 2C mice express an antigen receptor specific for the major histocompatibility complex class I alloantigen Ld and can be identified with the clonotypic monoclonal antibody 1B2. IT injection of lymphoid cells expressing Ld was found to induce a significant prolongation in BALB/c skin allograft survival. The allograft prolongation was associated with a marked reduction in the number of developing 1B2+ thymocytes (clonal deletion), which occurred primarily at the CD4+ CD8+ stage of thymocyte development, as well as a reduction in the number of mature CD8+ 1B2+ 2C T cells in peripheral lymphoid tissue. In addition, CD8+ 1B2+ 2C T cells that survive deletion have decreased CD8 expression levels and a significantly reduced in vitro proliferative response to specific alloantigen (clonal anergy). Exogenous recombinant interleukin 2 restores the capacity of 2C T cells to respond in vitro to alloantigen. Experiments involving separation of cells by fluorescence-activated cell sorter indicate that there is a precise correlation between the reduction in CD8 expression and anergy induction. Collectively, these data indicate that IT antigen injection can induce antigen-specific systemic tolerance by both clonal deletion and clonal anergy.

Intrathymic alloantigen-mediated, tolerant, completely major histocompatibility complex-mismatched mouse hearts are specifically rejected by adoptively transferred anti-class I L(d+)-specific 2C cells.

BACKGROUND: Tolerance to cardiac allografts can be induced in mice and rats by the injection of donor alloantigen into the thymus in combination with a CD4 T-cell-depleting antibody. CD8(+) cells in these animals are hyporesponsive to graft-specific alloantigens. Most of the CD8(+) T cells in the transgenic 2C mouse express a T-cell receptor specific for the class I major histocompatibility complex L(d+) locus. This study was designed to determine whether the adoptive transfer of these 2C T cells could precipitate rejection of a tolerant, completely major histocompatibility complex-mismatched L(d+) or L(d-) heart. METHODS: C57BL/6 mice (L(d-)) were given 10 x 10(6) cells of BALB/c (L(d+)) or dm2 (BALB/c background lacking L(d) [L(d-)]) splenocytes intrathymically and GK1. 5 (10 mg/kg) intraperitoneally. Twenty-one days later, BALB/c or dm2 hearts were transplanted. On the day of transplantation or after long-term allograft acceptance, recipients received naive 2C cells or 2C cells sensitized by in vitro mixed lymphocyte culture with BALB/c (L(d+)). RESULTS: Mean survival time of BALB/c cardiac allografts in untreated C57BL/6 mice was 7.3 days, although 73% of the mice that were pretreated with BALB/c splenocytes IT plus GK1.5 accepted the donor antigen-specific heart allografts indefinitely. All recipients that were pretreated with the intrathymic plus GK1.5 and that were injected with naive 2C cells at the time of heart transplantation experienced rejection of the BALB/c (L(d+)), but not the dm2 (L(d-)) hearts. In contrast, naive 2C cells could not reject tolerant (>30 days acceptance) BALB/c (L(d+)) hearts. 2C cells sensitized in vitro against L(d) were able to reject established BALB/c hearts but could not reject the L(d-) dm2 hearts. CONCLUSIONS: L(d)-specific 2C T-cell receptor transgenic T cells that are adoptively transferred to recipients will precipitate the rejection of accepted hearts that express class I L(d+) in mice rendered tolerant by an intrathymic injection of alloantigen plus anti-CD4 monoclonal antibodies.

Immunogenicity of L(d+) transgenic mouse hearts.

BACKGROUND: C57BL/6 mice transfected with the L(d) gene coupled to the alpha-myosin heavy chain promoter result in transgenic mice with L(d) antigen expressed only on cardiac tissue. These transgenic animals allow the examination of immune reactivity against cardiac L(d) by "self" or by adoptively transferred L(d) specific 2C cells, and the response of nontransgenic C57BL/6 mice to the transplanted L(d+) heart. METHODS: Naïve cardiac L(d+) transgenic mice were examined for evidence of L(d) "autoimmunity." Forty million fresh 2C cells or 2C cells sensitized in vitro for 7 days against Balb/c (L(d+)) + interleukin-2 were also given intravenously to L(d+) transgenic mice. At 5 and 12 days after injection, heart-infiltrating lymphocytes were analyzed by fluorescence-activated cell sorter. The L(d+) transgenic hearts were also transplanted to syngeneic L(d-) nontransgenic C57BL/6 to evaluate the heart's immunogenicity. RESULTS: Naïve L(d+) transgenic mice did not exhibit any evidence of lymphocytic infiltration on histologic examination. Adoptive transfer of either fresh or in vitro sensitized 2C cells was also unable to reject the native L(d+) heart in transgenic mice (100% of the mice survived long term [more than 60 days]). Sensitization of the L(d+) transgenic mice with a Balb/c skin graft and interleukin-2 pump infusion (7 days) beginning 1 day before 2C cell injection also did not promote rejection of the native L(d+) heart. However, fluorescence-activated cell sorter analysis did reveal that a significantly greater number of in vitro sensitized 2C cells homed to the L(d+), but not L(d-), heart after both 5 and 12 days (P <.01, P <.001). In contrast, C57BL/6 mice rejected the L(d+) (C57BL/6 background) transgenic heart in a mean survival time of 17 +/- 9.7 days (P <.01), whereas a syngeneic C57BL/6 heart transplant was accepted indefinitely. Lymphocytic infiltration consistent with rejection was present in all animals receiving an Ld+ transgenic heart transplant, whereas no infiltrate was present in those receiving a syngeneic C57BL/6 heart transplant. CONCLUSIONS: Although the class I L(d) transgene is not recognized in its native host, its immunogenicity is shown by the homing of anti-L(d) 2C cells to the heart in situ and rejection of L(d+) heart grafts when transplanted into syngeneic C57BL/6 mice.

Prenatal plasma matrix metalloproteinase-9 levels to predict spontaneous preterm birth.

OBJECTIVE: Matrix metalloproteinase-9, a zinc-dependent proteinase, may be important in initiating labor or rupture of membranes. We determined plasma levels of this enzyme in nonpregnant and pregnant women and evaluated whether they predict spontaneous preterm delivery. METHODS: A sandwich enzyme-linked immunosorbent assay (ELISA) was used to measure matrix metalloproteinase-9 levels in plasma samples from 25 nonpregnant women (mean age 39+/-9 years) and in stored plasma samples obtained during a randomized trial of zinc supplementation in pregnancy. Women were selected who delivered following spontaneous labor or premature rupture of membranes at 24-32 weeks (n = 20), 33-36 weeks (n = 29), and greater than 37 weeks (n = 30). Plasma samples were obtained sequentially at 19, 26, 31, and 36 weeks if undelivered and at presentation for delivery. RESULTS: Plasma matrix metalloproteinase-9 levels for non-pregnant women averaged 18.6+/-11.2 (mean +/- standard deviation) ng/mL. Prenatal values averaged 298+/-227 ng/mL from 19 weeks until 36 weeks (not including presentation for delivery) and did not change significantly as the gestational age increased, regardless of whether women ultimately delivered at 24-32, 33-36, or after 37 weeks. Values obtained prior to, but within 1 week of, presentation for delivery (n = 7, 281+/-103 ng/mL) were not significantly different than those obtained earlier in pregnancy (n = 71, 309+/-307 ng/mL, [P = .60]). Plasma matrix metalloproteinase-9 levels for women in spontaneous labor were similar regardless of gestational age and were increased three-fold (852+/-301 ng/mL) compared with those drawn at each prenatal visit (for example, 26 week values = 285+/-144 ng/mL [P < .001]). CONCLUSION: Plasma matrix metalloproteinase-9 levels remain unchanged throughout pregnancy until the onset of spontaneous labor when there is a three-fold increase. Plasma matrix metalloproteinase-9 levels obtained prior to presentation for delivery do not appear to predict spontaneous preterm birth.