Tail Swelling Research Articles

Tissue-Specific Activities of an Immune Signaling Module Regulate Physiological Responses to Pathogenic and Nutritional Bacteria in C. elegans

Microbes represent both an essential source of nutrition and a potential source of lethal infection to the nematode Caenorhabditis elegans. Immunity in C. elegans requires a signaling module comprised of orthologs of the mammalian Toll-interleukin-1 receptor (TIR) domain protein SARM, the mitogen-activated protein kinase kinase kinase (MAPKKK) ASK1, and MAPKK MKK3, which activates p38 MAPK. We determined that the SARM-ASK1-MKK3 module has dual tissue-specific roles in the C. elegans response to pathogens--in the cell-autonomous regulation of innate immunity and the neuroendocrine regulation of serotonin-dependent aversive behavior. SARM-ASK1-MKK3 signaling in the sensory nervous system also regulates egg-laying behavior that is dependent on bacteria provided as a nutrient source. Our data demonstrate that these physiological responses to bacteria share a common mechanism of signaling through the SARM-ASK1-MKK3 module and suggest the co-option of ancestral immune signaling pathways in the evolution of physiological responses to microbial pathogens and nutrients.

Transient Anti-GAD Antibody Positivity and Acute Pancreatitis with Pancreas Tail Swelling in a Patient with Susceptible Haplotype for Type 1 Diabetes Mellitus

A 57-year-old man was admitted to our hospital complaining of poor appetite. He had been diagnosed with diabetes mellitus and was anti-GAD antibody (GAD-Ab) negative 1 year previously, at the age of 56 years old. Abdominal CT revealed pancreas tail swelling; elastase-I level was elevated and he was diagnosed with pancreatitis. The level of GAD-Ab was increased and HLA haplotype was DRB1*0901-DQB1*0303, which is seen frequently in type 1 diabetic Japanese patients. However, his endogenous insulin secretion ability was not deteriorated. After elastase-I level and pancreas swelling improved, GAD-Ab returned to a normal range. One year after the onset of pancreas swelling, he was still not in an insulin-dependent state. In this case, transient GAD-Ab positivity with susceptible haplotype for type 1 diabetes mellitus might have been induced by a GAD antigen discharged from the destroyed islet due to pancreatitis.

Evaluation of acrosomal status and sperm viability in fresh and cryopreserved specimens by the use of fluorescent peanut agglutinin lectin in conjunction with hypo-osmotic swelling test

In this study, we evaluated whether the hypo-osmotic swelling test (HOST) can be used as a vital marker in combination with peanut agglutinin (PNA) - labeling in fresh and cryopreserved spermatozoa. Human sperm populations were exposed to a hypo-osmotic medium for 60 minutes, and then incubated in a 1 microg/mL solution of the fluorescent dye Hoescht 33258 (H33258) for 10 minutes. Excess stain was removed by washing in phosphate-buffered saline (PBS) solution, and the pellet was resuspended in 100 microL of culture medium. Twenty microliters of this solution were subsequently smeared on a microscope slide, and fixed in ice-cold methanol to permeabilize the sperm membranes. The fixed smears were finally incubated in a 40-microg/mL FITC-PNA solution for 20 minutes. Simultaneous assessment of acrosome and viability scores was done in a fluorescent microscope equipped with appropriate filters and phase contrast illumination. The same slide was examined for FITC-PNA labeling, tail swelling, and for Hoechst-33258 staining by interchanging the filters and phase contrast optics. In fresh specimens, HOST was found to provide viability assessments comparable to those obtained using the H33258 method (r = 0.95). However, the results of HOST and H33258 were not correlated in cryopreserved specimens (r = 0.22). There was no alteration of PNA-labeling due to the HOST or H33258. FITC-PNA labeling in conjunction with the visualization of the morphological change induced by exposure to hypo-osmotic solution provides a simple but effective method for establishing the state of acrosomal membrane and viability in fresh human spermatozoa, but this technique is not reliable for cryopreserved ones.

개에서 Pentoxifylline 첨가에 따른 동결정액 성상과 인공수정에 관한 연구

본 연구는 개 정액 채취방법에 따른 정액성상과 동결전 pentoxifylline의 첨가가 미치는 영향을 조사하였고, CASA를 이용하여 정자의 운동성 측정 및 생존율, 첨체 온전성, 저삼투성 팽창화를 조사하였다. 그리고 자연발정과 발정유기된 암캐에서 동결정액 주입의 임신율을 조사하였다. 1.품종별 정액성상을 조사한 결과 Beagle, English cockers spanie, Collie 및 Grey hound의 정액량과 정자 수에서 큰 차이가 없었으나, Shihtzu에서 정액량과 정자수가 유의적은 아니나 다소 적었다. 2.동결희석액에 pentoxifylline 0, 1 및 3mM 첨가에서 융해 후 정자활력은 각각 46.4, 56.4 및 47.2%였으며 1mM 처리구가 다른 처리구보다 유의적으로 높았다. 3.동결희석액에 pentoxifylline 첨가는 융해 후 정자의 생존성, 첨체막 온전성 및 원형질막 온전성(HOST) 모두 유리한 결과를 나타냈으며, 1mM의 첨가에서 정자의 생존성과 기능성이 향상되었다.4.자연발정에서 발정개시 후 11일, 13일 및 15일째 초음파기로 측정한 난포 크기는 각각 6.5mm, 11.5mm 및 8.0mm이였다. 배란은 13-15일 사이에 일어났다. 5.발정유기한 개체에서 수정후 임신 20일째부터 5일 간격으로 3회 측정한 태낭의 크기는 13.7mm, 28.5mm 및 40.5mm 이었다. 6.자연발정과 발정을 유기한 개체에서 pen- toxifylline 1mM이 첨가된 정액을 동결융해하여 인공수정 후 임신율은 71.4%와 75%로 두 방법간에 차이가 없었으나, 자연발정군에서 평균산자수는 6.6두였고, 발정유군의 평균산자수는 2.6두였다. The present study evaluated whether pentoxifylline added to the freezing extender could improve the sperm characteristics and function in canine frozen semen. Also the conception rate following AI with frozen-thawed semen was investigated. The beneficial effects of pentoxifylline supplementation were visible in motility, viability, acrosome reaction, and tail swelling patterns. Especially, highest sperm viability and function were obtained in the forzen semen supplemented with 1mM pentoxifylline. The follicle size measured by ultrasonography was 6.48 mm, 11.52 mm and 8.9 mm on 11, 13 and 15 days after the onset of natural estrus, respectively and ovulation occurred on 13 and 15 days. The pregnancy rates in bitches inseminated with frozen semen on natural and induced estrus were 71.4% and 75.0%, respectively. There was no significant difference between the pregnancy rates in bitches inseminated with frozen semen following natural and induced estrus, but the litter size was slightly increased in natural cycle.

Multiple Genes Affect Sensitivity of Caenorhabditis elegans to the Bacterial Pathogen Microbacterium nematophilum

Interactions with bacteria play a major role in immune responses, ecology, and evolution of all animals, but they have been neglected until recently in the case of C. elegans. We report a genetic investigation of the interaction of C. elegans with the nematode-specific pathogen Microbacterium nematophilum, which colonizes the rectum and causes distinctive tail swelling in its host. A total of 121 mutants with altered response to infection were isolated from selections or screens for a bacterially unswollen (Bus) phenotype, using both chemical and transposon mutagenesis. Some of these correspond to known genes, affecting either bacterial adhesion or colonization (srf-2, srf-3, srf-5) or host swelling response (sur-2, egl-5). Most mutants define 15 new genes (bus-1-bus-6, bus-8, bus-10, bus-12-bus-18). The majority of these mutants exhibit little or no rectal infection when challenged with the pathogen and are probably altered in surface properties such that the bacteria can no longer infect worms. A number have corresponding alterations in lectin staining and cuticle fragility. Most of the uninfectable mutants grow better than wild type in the presence of the pathogen, but the sur-2 mutant is hypersensitive, indicating that the tail-swelling response is associated with a specific defense mechanism against this pathogen.

Hypoosmotic swelling patterns in human spermatozoa: Effects of colloid osmotic pressure conditions

Objective: Addition of proteins to hypoosmotic media delays the swelling response as compared to hypoosmotic solutions without protein supplementation. The hypoosmotically-reactive spermatozoa are able to maintain motility, and even progressive motility without associated tail swelling, in the presence of proteins in solution. The aim of this study was to assess the sperm response at the onset of swelling, and also during the development of the tail swelling patterns until reaching their optimal response under hypoosmotic conditions regulated by the presence of proteins.

Rheologic Behavior of Spermatozoa Exposed to Hypoosmotic Conditions at Varying Oncotic Pressure Environments: Possible Protective Mechanisms Against Sperm Swelling by the Addition of Protein Supplements

Objective: Exposure of spermatozoa to hypoosmotic conditions that cause optimal swelling for assessment of sperm membrane functional integrity causes sudden and irreversible changes in membrane structure. In almost all cases, spermatozoa cannot support viability as observed by lack of motility during and at the end of the incubation period (30 to 60 min). This behavior of human spermatozoa exposed to hypoosmotic conditions has been previously assessed, which resulted in the development of a sperm functional test (hypoosmotic swelling (HOS) test. J Reprod Fertil 1984;70:219). The aim of this study was to assess possible osmoregulatory mechanisms of substances contributing to oncotic pressure (exerted by colloids in solution) such as protein supplements against exposure to hypoosmotic conditions (exerted by dissolved substances in solution). Design: Exposure of processed spermatozoa to controlled hypoosmotic conditions (150 mOsm/L) with varying serum supplement (Synthetic serum substitute; SSS, Irvine Scientific) concentrations contributing to variability in the oncotic pressure environment. Stock solutions consisted of HOS solution and hypoosmotic SSS (H-SSS) at 150 mOsm/L. The various stock and mixtures of solutions tested consisted of: 1) HOS solution 100%, 2) H-SSS 100%, 3) 75% H-SSS/25% HOS, 4) 50% H-SSS/50% HOS, and 5) 25% H-SSS/75% HOS. Materials and Methods: Semen specimens from 18 men were obtained for routine semen analysis and pooled (2 to 3 specimens each time). Pooled specimens were processed via density gradient centrifugation and reconstituted in human tubal fluid (HTF; Irvine Sci) media containing 0% SSS. Aliquots of the reconstituted specimens were diluted 1:10 (v/v) with each of the testing solutions and incubated for 60 min (37°C/5% CO2). Specimens were assessed on wet mounts for total swelling and swelling patterns, which included maximal (>50% tail swelling) and minimal swelling (<50% tail swelling), and motile sperm undergoing swelling of any type. Results: The results obtained are shown in the table below: legendType of swellinglegend∗, †, ‡, §, ∥. Denotes significant differences (P<0.05) between rows.H-SSS 100%75% H-SSS/ 25% HOS50% H-SSS/ 50% HOS25% H-SSS/ 75% HOSHOS 100%Maximal swelling13.4% ± 2.0∗20.6% ± 1.7†35.9% ± 1.7‡43.1% ± 2.7§50.8% ± 2.0∥Minimal swelling23.9% ± 1.6∗34.2% ± 1.8†33.3% ± 1.5†33% ± 1.8†27.9% ± 1.6‡Motile/swelling51.4% ± 2.5∗31.8 ± 2.3†15.4% ± 1.2‡6.3% ± 1.0§1.1% ± 0.4∥Total swelling88.8%86.6%84.7%82.3%79.8%legend ∗, †, ‡, §, ∥. Denotes significant differences (P<0.05) between rows. Open table in a new tab Conclusions: The addition of increasing concentrations of protein supplements resulted in a decrease in the distribution of maximal swelling patterns and an increase in the proportion of minimal swelling and motile spermatozoa undergoing minimal swelling. Furthermore, the overall sperm swelling patterns increased with an increase in the protein supplement concentration. Thus, addition of serum supplements may be beneficial to provide protection against changes in osmotic pressure in culture media, but caution is advised when processing spermatozoa for freezing or thawing due to the possible interaction between protein molecules and free-water, which freezes or thaws first, causing the opposite effect of further increasing the osmotic pressure of the medium.

Effects of calcium pentosan polysulfate on joint inflammation and pouch fluid levels of leukocytes, nitric oxide, and interleukin-6 in a rat model of arthritis

Abstract This study evaluated the antiarthritic and anti-inflammatory activities of calcium pentosan polysulfate (CaPPS), using a modified adjuvant-induced arthritis model in rats. A combined model of arthritis and air-pouch inflammation was developed in Wistar rats by subdermal injection of Mycobacterium tuberculosis (Mtb) into the tail base on day 0 followed by a second Mtb injection on day 10 into preformed, saline-filled, dorsal air pouches. Rats received either normal saline only (n=25) or CaPPS in saline administered subcutaneously at doses of 2.5 (n=5), 5 (n=5), 10 (n=5), or 20 mg/kg (n=10) 5 times per week over the study period. Animals were necropsied on day 17, and pouch fluid volume, total and differential white blood cell (WBC) counts, nitrite ion (the metabolite of the nitric oxide [NO] radical), prostaglandin E 2 (PGE 2 ), and interleukin-6 (IL-6) content were determined. In addition, IL-6 activity in serum samples collected at necropsy was evaluated. Results of this study showed that CaPPS at 10 and 20 mg/kg significantly reduced tail and paw swelling ( P 2 levels, it reduced total WBC counts ( P P 2.5 mg/kg ( P 2 levels in inflamed subcutaneous rat air-pouch fluids.

Phototoxicity of quinalphos under sunlight in vitro and in vivo

The phototoxicity of quinalphos (QP) under sunlight was studied by investigating skin sensitization, mouse tail and ear swelling tests and generation of activated oxygen species (AOS). Quinalphos, at an oral dose of 5 mg/kg to the mouse, did not produce any change in water content of the tail, but produced ear swelling under sunlight (60 min). The pesticide did not produce any skin sensitization reaction when applied on depilated skin of guinea pigs (100–500 μg/cm 2) and irradiated under sunlight (60 min). Furthermore, the compound produced a significant amount of AOS such as singlet oxygen ( 1O 2) and superoxide anion radical ( O - 2 ̇ under sunlight. The production of AOS was both concentration (1–20 μg/ml) and irradiation time (0–40 min)-dependent. The generation of AOS was also confirmed by using quenchers of singlet oxygen (sodium azide and DABCO) and superoxide anion radical (SOD).

Effects of hypoosmotic incubation on acrosome and tail structure on canine spermatozoa

Hypoosmotic tests are widely used as valuable tests for determining sperm quality in species as varied as the human and the porcine. However, there is little information about the use of these tests in canine spermatozoa. This work evaluates the response of canine spermatozoa in hypoosmotic media in order to introduce the use of the hypoosmotic tests in the canine standard semen analysis. In this way, the incubation of canine spermatozoa in hypoosmotic media containing citrate (ORT medium, osmotic pressure = 100 mOsm) or citrate plus fructose (HOS medium, osmotic pressure = 150 mOsm) resulted in the swelling of the sperm tail. These reactions were time-dependent, reaching maximum percentages after 45 to 60 min. Optimal percentage of tail swelling with minimal effect on the viability of spermatozoa was observed at 100 to 150 mOsm. Response on sperm viability, tail swelling and acrosome detachment to hypoosmotic tests of both undiluted fresh, and 24 h-stored samples were similar. The percentage of swollen tails after both tests showed a good correlation to viability and to gross and progressive motility but not to concentration. However, acrosome detachment after both hypoosmotic tests did not correlate to any of the studied parameters. Our results indicate that the swelling observed after hypoosmotic shock could be used as a useful test in improving the standard semen analysis in the dog.

Divergence between delayed-type hypersensitivity (DTH) to polyoma tumor-associated antigen and antitumor efficacy in polyoma-bearing recipients of therapeutic non-cytolitic, DTH-mediating lymphocytes

Soluble polyoma tumor-associated antigen (TAA) restimulation of splenocytes from polyoma-bearing mice led to production of non-cytolytic lymphocytes of helper-cell phenotype which after i.p. administration mediate inhibition of polyoma and delayed-type hypersensitivity (DTH) to polyoma TAA. The maximum efficacy of the two activities was found in cells isolated on day 9 when also the highest percentage of L3T4 + cells was measured. The different rates of decline along with the extended life of the culture, and also the different degrees of dependence on the recipient's immune system, indicate that these processes are independent. The range of tail swelling in tumor-bearing recipients of immunotherapy reflects only the DTH transfer activity and not the range of cooperation between the recipient's immune system and the transferred T-helper lymphocytes leading to in vivo generation of CTL.

Human spermatozoal tail hypo-osmotic swelling test, motility characteristics in hypotonic saline, and survival of spermatozoa after cryopreservation.

Normozoospermic semen samples (n = 82) were examined to investigate whether the degree of sperm tail swelling in hypo-osmotic medium (fructose and sodium citrate; 150 mOsm/l), and motility characteristics after a 15-min exposure to hypotonic saline (sodium chloride; 150 mOsm/l) could predict the cryosurvival rate (% post-thaw motility/% pre-freeze motility x 100%) of spermatozoa after cryopreservation by the liquid nitrogen vapour freezing method using the TEST-glycerol-egg yolk buffer. The CellSoft automated semen analyser was used to analyse sperm motility in pre-freeze and post-thaw semen samples, and after exposure to hypotonic saline. Sperm tail hypo-osmotic swelling and sperm motility in pre-freeze semen showed no significant correlations (P > 0.05) with the cryosurvival rate. There were significant correlations (P < 0.05) between the cryosurvival rate and the following sperm motility parameters in hypotonic saline: % motility (r = 0.2846), motility index (% motility x curvilinear velocity; r = 0.2809) and % decrease in motility index from the baseline value in semen (r = 0.3378). The % decrease in motility index after hypotonic saline treatment was significantly less (P < 0.05) in the normal (> or = 50% cryosurvival rate; mean +/- SEM 5.9 +/- 3.2%; n = 33) compared with the subnormal (< 50% cryosurvival rate; 27.3 +/- 4.8%; n = 49) cryosurvival groups. This parameter was also determined, by multivariate discriminant analysis, to be capable of classifying each pre-freeze semen sample into either cryosurvival group with 69.5% accuracy.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of leucocytes on the hypoosmotic swelling test of human sperm.

The hypoosmotic swelling test was used to evaluate the size of physiologically competent sperm after their exposure to the influence of leucocyte-derived superoxide anions. In sperm suspensions, preincubated in the presence of 50 x 10(4)mL-1 leucocytes capable of producing 0.93 +/- 0.63 nmol of O2- (range 0.21-2.03 nmol) over 30 min at 37 degrees C, a significant decrease in the percentage of swollen sperm compared to that in the control sperm suspensions without added leucocytes was found (p < .001, n = 46). Addition of phorbol myristate acetate, an activator of the release of O2-, further diminished the ability of hypoosmotic sperm tail swelling (p < .001), while superoxide dismutase inhibited this effect. This study suggests that even very low amounts of superoxide anions may influence the functional integrity of sperm. Therefore, any source of superoxide anion production should be eliminated and/or inhibited in sperm suspensions used in assisted reproduction techniques.

Reappraisal of the hypo-osmotic swelling test to improve assessment of seminal fertility status.

The hypo-osmotic swelling (HOS) test has been proposed as a useful assay for evaluation of the functional competence of the human sperm membranes. To assess this further, the HOS-test was evaluated in 187 semen samples collected from fertile men and from male patients consulting for infertility. These samples were classified as normal, oligo-, astheno- or oligoasthenozoospermic on the basis of their standard semen variables. The percentage of total sperm tail swelling and of sperm exhibiting different tail swelling patterns was recorded. In the fertile men and in the group of patients with normal semen variables, significantly more (P less than 0.001) HOS-reactive sperm were observed after hypo-osmotic treatment in comparison with those groups exhibiting abnormal semen parameters. Swelling of the sperm in a hypo-osmotic medium was highly correlated with both progressive motility (r = 0.62, P less than 0.001) and sperm viability (r = 0.65, P less than 0.001). A weak positive correlation was also observed between sperm swelling and sperm morphological features (r = 0.31, P less than 0.005) and between sperm swelling and sperm concentration (r = 0.31, P less than 0.005). No significant correlation was observed between sperm swelling and in-vitro sperm fertilizing capacity as assessed by the zona-free hamster oocyte penetration assay. However the majority of the semen samples (87.3%) showing a normal penetration rate (greater than or equal to 10%) also exhibited a 60% (or higher) reaction in the HOS-test.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of computerized analysis of sperm movement characteristics and differential sperm tail swelling patterns in predicting human sperm in vitro fertilizing capacity.

Multivariate discriminant analysis was used to evaluate the usefulness of computer image analysis of sperm movement characteristics and differential patterns of sperm tail swelling after hypoosmotic treatment for predicting the human sperm in vitro fertilizing capacity assessed by the zona-free hamster egg penetration assay. Fifty-five semen samples, mostly normospermic, from untreated infertility clinic patients were analyzed. The % normal sperm morphology, linearity of seminal sperm movement, seminal sperm head beat frequency, mean and maximum amplitudes of lateral head displacement, and hypoosmotic sperm tail swelling patterns c, d and f were selected by multivariate discriminant analysis to be capable of discriminating the samples exhibiting the presence or the absence of sperm in vitro fertilizing capacity. The % total sperm tail swelling did not give additional information about in vitro fertilizing capacity. These preliminary data suggest that computer image analysis of sperm movement characteristics and differential evaluation of hypoosmotic sperm tail swelling might be useful for the prediction of human sperm fertility. Further prospective studies are necessary to validate their predictive functions.

A study of a magnetospheric substorm recorded by the Viking auroral imager

A substorm, recorded at 80 second time resolution in images from the Viking auroral imager, has been studied using data from ground observations, from a geosynchronous satellite and from ISEE 1 in the magnetotail plasma sheet. Substorm onset, marked by negative bays at the ground, particle injection at synchronous orbit and plasma sheet dropout at ISEE 1, appeared to produce only local brightening of the night side oval. A rapid poleward expansion of the auroras twenty minutes later was accompanied by a major reduction of tail energy density and swelling of the inner plasma sheet. Recovery, 15 minutes later, of auroral zone bays and the plasma sheet was accompanied by fading of the equatorward auroras that left a bright poleward arc as the primary auroral emission.

Protection of mice against vaccinia virus by bacterial infection and sustained stimulation with specific bacterial antigens.

In these experiments, mice which have a strong delayed-type hypersensitivity to mycobacteria were found, when elicited with old tuberculin, to be more resistant to intravenous vaccinia virus challenge than controls. This was manifest as protection from killing when large amounts of virus were injected, or as significantly less tail swelling and damage as well as lower titers of infectious virus when a lesser inoculum was used. Preliminary experiments indicate that animals sensitized with Staphylococcus aureus and elicited with phage lysate of staphylococcus are also more resistant to vaccinia infection.