Tail Swelling Research Articles

Ultrasound Irradiation as a Candidate Procedure to Improve the Transdermal Drug Delivery to the Tail Edema of a Mouse Model

Drug therapy for secondary lymphedema has not yet been established. Conventional oral and intravenous administration is difficult to administer in sufficient doses due to adverse events. Therefore, it is necessary to develop a transdermal delivery system that can deliver high concentrations of drugs to the edema area. In this study, we examined the efficacy of transdermal drug delivery in a mouse model of tail edema using ultrasound irradiation (sonication method). Ultrasound irradiation can deliver high-molecular-weight substances subcutaneously, and the percutaneous administration of clobetasol propionate to the mouse tail edema model prevented the enlargement of lymphatic vessels with reduced tail volume. Therefore, steroid administration utilizing ultrasound irradiation is effective in decreasing tail swelling in a mouse tail edema model. Thus, ultrasound irradiation could have the potential to innovate the treatment of secondary lymphedema by directly administering the drug to the edema.

Pannexin1 deletion in lymphatic endothelium affects lymphatic function in a sex-dependent manner.

The lymphatic network of capillaries and collecting vessels ensures tissue fluid homeostasis, absorption of dietary fats and trafficking of immune cells. Pannexin1 (Panx1) channels allow for the passage of ions and small metabolites between the cytosol and extracellular environment. Panx1 channels regulate the pathophysiological function of several tissues in a sex-dependent manner. Here, we studied the role of Panx1 in lymphatic function, and potential sex-dependent differences therein, in Prox1-CreERT2Panx1fl/fl and Panx1fl/fl control mice. Panx1 expression was higher in lymphatic endothelial cells (LECs) of male mice. Lymphatic vessel morphology was not affected in Prox1-CreERT2Panx1fl/fl male and female mice. Lymphatic drainage was decreased by 25% in male Prox1-CreERT2Panx1fl/fl mice, but was similar in females of both genotypes. Accordingly, only male Prox1-CreERT2Panx1fl/fl mice exhibited tail swelling, pointing to interstitial fluid accumulation in males upon Panx1 deletion in LECs. Moreover, serum triglyceride and free fatty acid levels raised less in Prox1-CreERT2Panx1fl/fl mice of both sexes in an oral lipid tolerance test. Finally, the percentage of migratory dendritic cells arriving in draining lymph nodes was increased in Prox1-CreERT2Panx1fl/fl female mice, but was comparable between male mice of both genotypes. Our results point to a LEC-specific role for Panx1 in the functions of the lymphatic system.

A Look Inside-Histopathological Examinations of Different Tail Tip Lesions in Dairy Cows.

Feedlot cattle are frequently affected by inflammation and necrosis of the tail tips, resulting in impeded animal welfare and economic losses. In a recent study, it was demonstrated that dairy cows are also affected by different lesions of the tail tip, including alopecia, annular constrictions, crusting, scaling and swelling. Despite the frequent occurrence of these lesions, the underlying etiology and pathomechanisms are unclear to date. To gain insight into this malady, we histopathologically examined 16 tail tips of slaughtered dairy cows, representing the entire range of different lesions. While macroscopically alopecic areas were characterized by the formation of granulation tissue in the dermis with an inconspicuous epidermis, we found not only dermal granulation tissue but also purulent-necrotizing inflammation with the breakdown of the basement membrane in encrusted lesions. Interestingly, in some cases, we found areas of coagulation necrosis of the epidermal and dermal layers in the crusts. Tails with macroscopical scaling were affected by ortho- and/or parakeratotic hyperkeratosis, and, in tails with macroscopical swelling, we observed a low-protein edema of the central longitudinal connective tissue of the tail. We conclude that the observed lesions might be caused by ischemia of the skin in the distal parts of the tail.

Abnormal Lymphatic Sphingosine-1-Phosphate Signaling Aggravates Lymphatic Dysfunction and Tissue Inflammation.

Lymphedema is a global health problem with no effective drug treatment. Enhanced T-cell immunity and abnormal lymphatic endothelial cell (LEC) signaling are promising therapeutic targets for this condition. Sphingosine-1-phosphate (S1P) mediates a key signaling pathway required for normal LEC function, and altered S1P signaling in LECs could lead to lymphatic disease and pathogenic T-cell activation. Characterizing this biology is relevant for developing much needed therapies. Human and mouse lymphedema was studied. Lymphedema was induced in mice by surgically ligating the tail lymphatics. Lymphedematous dermal tissue was assessed for S1P signaling. To verify the role of altered S1P signaling effects in lymphatic cells, LEC-specific S1pr1-deficient (S1pr1LECKO) mice were generated. Disease progression was quantified by tail-volumetric and -histopathologic measurements over time. LECs from mice and humans, with S1P signaling inhibition, were then cocultured with CD4 T cells, followed by an analysis of CD4 T-cell activation and pathway signaling. Last, animals were treated with a monoclonal antibody specific to P-selectin to assess its efficacy in reducing lymphedema and T-cell activation. Human and experimental lymphedema tissues exhibited decreased LEC S1P signaling through S1P receptor 1 (S1PR1). LEC S1pr1 loss-of-function exacerbated lymphatic vascular insufficiency, tail swelling, and increased CD4 T-cell infiltration in mouse lymphedema. LECs, isolated from S1pr1LECKO mice and cocultured with CD4 T cells, resulted in augmented lymphocyte differentiation. Inhibiting S1PR1 signaling in human dermal LECs promoted T-helper type 1 and 2 (Th1 and Th2) cell differentiation through direct cell contact with lymphocytes. Human dermal LECs with dampened S1P signaling exhibited enhanced P-selectin, an important cell adhesion molecule expressed on activated vascular cells. In vitro, P-selectin blockade reduced the activation and differentiation of Th cells cocultured with shS1PR1-treated human dermal LECs. P-selectin-directed antibody treatment improved tail swelling and reduced Th1/Th2 immune responses in mouse lymphedema. This study suggests that reduction of the LEC S1P signaling aggravates lymphedema by enhancing LEC adhesion and amplifying pathogenic CD4 T-cell responses. P-selectin inhibitors are suggested as a possible treatment for this pervasive condition.

Effect of a dietary supplement on the reduction of lymphedema-progression in mouse tail-cut model.

The aim of our study was to evaluate in vivo, in a mouse tail model of lymphedema, the effects of a dietary supplement, Garlive®, based on hydroxytyrosol from olive leaves, spermidine from rice seeds, hesperidin from citrus fruits and vitamin A. Hydroxytyrosol has anti-inflammatory, antioxidant and antimicrobial activities and inhibits leukotriene B4 generation; spermidine is able to inhibit the production of pro-inflammatory cytokines and mediators; hesperidin inhibits the secretion of pro-inflammatory cytokines: IFN-γ, IL-2, IL-4, IL-10; vitamin A deficiency was shown to induce inflammation and aggravate existing inflammatory states, whereas supplementation with vitamin A could ameliorate inflammation. The active compounds were included in tablets: 250 mg of olive leaf extract titrated in 10% hydroxytyrosol, 200 mg of citrus fruits extract titrated in 60% hesperidin, 10 mg of rice (Oryza sativa) seeds extract titrated in 1% spermidine and 0.8 mg of vitamin A. Mice of an inbred group were randomly selected and divided in the control group and drug-treated group. The wound necessary for lymphedema generation was made on the tail of each mice 1 cm below the base of the trunk. After surgical intervention, there was a gradual increase in the circumference of both ends of the wound. The control group showed higher increase of tail volume than the drug-treated group. The differences in tail swelling between the control group and the drug-treated group were significantly different. The peak of swelling was anticipated to the 6th day in the drug-treated group, whereas in the control group the peak was reached later on. The tested drug prevented the induction of swelling from day 5th of wound creation and decreased the duration of swelling, favoring the wound healing.

Changes in tail posture detected by a 3D machine vision system are associated with injury from damaging behaviours and ill health on commercial pig farms.

To establish whether pig tail posture is affected by injuries and ill health, a machine vision system using 3D cameras to measure tail angle was used. Camera data from 1692 pigs in 41 production batches of 42.4 (±16.6) days in length over 17 months at seven diverse grower/finisher commercial pig farms, was validated by visiting farms every 14(±10) days to score injury and ill health. Linear modelling of tail posture found considerable farm and batch effects. The percentage of tails held low (0°) or mid (1–45°) decreased over time from 54.9% and 23.8% respectively by -0.16 and -0.05%/day, while tails high (45–90°) increased from 21.5% by 0.20%/day. Although 22% of scored pigs had scratched tails, severe tail biting was rare; only 6% had tail wounds and 5% partial tail loss. Adding tail injury to models showed associations with tail posture: overall tail injury, worsening tail injury, and tail loss were associated with more pigs detected with low tail posture and fewer with high tails. Minor tail injuries and tail swelling were also associated with altered tail posture. Unexpectedly, other health and injury scores had a larger effect on tail posture- more low tails were observed when a greater proportion of pigs in a pen were scored with lameness or lesions caused by social aggression. Ear injuries were linked with reduced high tails. These findings are consistent with the idea that low tail posture could be a general indicator of poor welfare. However, effects of flank biting and ocular discharge on tail posture were not consistent with this. Our results show for the first time that perturbations in the normal time trends of tail posture are associated with tail biting and other signs of adverse health/welfare at diverse commercial farms, forming the basis for a decision support system.

The Kinetics of Lymphatic Dysfunction and Leukocyte Expansion in the Draining Lymph Node during LTB4 Antagonism in a Mouse Model of Lymphedema.

The mechanisms of lymphedema development are not well understood, but emerging evidence highlights the crucial role the immune system plays in driving its progression. It is well known that lymphatic function deteriorates as lymphedema progresses; however, the connection between this progressive loss of function and the immune-driven changes that characterize the disease has not been well established. In this study, we assess changes in leukocyte populations in lymph nodes within the lymphatic drainage basin of the tissue injury site (draining lymph nodes, dLNs) using a mouse tail model of lymphedema in which a pair of draining collecting vessels are left intact. We additionally quantify lymphatic pump function using established near infrared (NIR) lymphatic imaging methods and lymph-draining nanoparticles (NPs) synthesized and employed by our team for lymphatic tissue drug delivery applications to measure lymphatic transport to and resulting NP accumulation within dLNs associated with swelling following surgery. When applied to assess the effects of the anti-inflammatory drug bestatin, which has been previously shown to be a possible treatment for lymphedema, we find lymph-draining NP accumulation within dLNs and lymphatic function to increase as lymphedema progresses, but no significant effect on leukocyte populations in dLNs or tail swelling. These results suggest that ameliorating this loss of lymphatic function is not sufficient to reverse swelling in this surgically induced disease model that better recapitulates the extent of lymphatic injury seen in human lymphedema. It also suggests that loss of lymphatic function during lymphedema may be driven by immune-mediated mechanisms coordinated in dLNs. Our work indicates that addressing both lymphatic vessel dysfunction and immune cell expansion within dLNs may be required to prevent or reverse lymphedema when partial lymphatic function is sustained.

A Murine Tail Lymphedema Model.

Lymphedema is extremity swelling caused by lymphatic dysfunction. The affected limb enlarges because of accumulation of fluid, adipose, and fibrosis. There is no cure for this disease. A mouse tail model that uses a focal full thickness skin excision near the base of the tail, resulting in tail swelling, has been used to study lymphedema. However, this model may result in vascular comprise and consequent tail necrosis and early tail swelling resolution, limiting its clinical translatability. The chronic murine tail lymphedema model induces sustained lymphedema over 15 weeks and a reliable perfusion to the tail. Enhancements of the traditional murine tail lymphedema model include 1) precise full thickness excision and lymphatic clipping using a surgical microscope, 2) confirmation of post-operative arterial and venous perfusion using high resolution laser speckle, and 3) functional assessment using indocyanine green near infrared laser lymphangiography. We also use tissue nanotransfection technology (TNT) for novel non-viral, transcutaneous, focal delivery of genetic cargo to the mouse tail vasculature.

Sensitivity and specificity of a tail-activity measuring device for calving prediction in dairy cattle

Efficient calving surveillance is essential for avoiding stillbirth due to unattended dystocia. Calving sensors can help detect the onset of parturition and thus ensure timely calving assistance if necessary. Tail-raising is an indicator of imminent calving. The objective of this study was to evaluate a tail-mounted inclinometer sensor (Moocall Ltd., Dublin, Ireland) and to monitor skin integrity after sensor attachment. Cows (n = 157) and heifers (n = 23) were enrolled at 275 d post insemination, and a sensor was attached to each cow's tail. Investigators checked for signs indicating the onset of stage II of parturition, verified the position of the sensor, and evaluated the skin integrity of the tail above and below the sensor hourly for 24 h/d. We used 5 different intervals (i.e., 1, 2, 4, 12, and 24 h until calving) to calculate sensitivity and specificity. Sensors continuously remained on the tail (i.e., within 3 cm of the initial attachment position) after initial attachment until the onset of calving in only 13.9% of animals (n = 25). Sensors were reattached until a calving event occurred (51.6%) or the animal was excluded for other reasons (34.4%). In 31 animals the sensor was removed because the tail was swollen or painful. Heifers were significantly less likely than cows to lose a sensor but more likely to experience tail swelling or pain. Depending on the interval preceding the onset of parturition, sensitivity varied from 19 to 75% and specificity from 63 to 96%.

Effects of the Traditional Chinese Medical Prescription Linba Fang as a Treatment for Lymphedema

Lymphedema can lead to a series of complicated and irreversible chronic pathological changes, including lymphatic fluid retention, infiltration of inflammatory cells, lipid deposition, and fibrosis of the surrounding tissues. Typically, compression physiotherapy is recommended for early lymphedema. However, the chronic fluid compartments will lead to fat deposition, skin fibrosis, and hyperkeratosis. Few treatment methods are available for patients with lymphedema. Previous studies have attempted to apply diuretics, diosmin, and sodium β-aescinate to treatment for venous edema, but the curative effect was unsatisfactory. There is currently no established effective treatment for lymphedema. In this paper, we investigated the effects of the traditional Chinese medical prescription Linba Fang as a treatment for lymphedema using a mouse model. A lymphedema model was established in C57BL/6 mice through lymphatic ablation at the base of tails. Negative controls were administered with 0.5% sodium carboxymethyl cellulose solution by gavage twice daily, positive controls with aescuvenforte, and test mice with Linba Fang. Aescuvenforte and Linba Fang were dissolved in 0.5% sodium carboxymethyl cellulose solution to produce a homogeneous mixture. After treatment for 2–4 weeks, tail diameter and weight, inflammatory cytokine levels (IL-1, IL-6, and TNF-α), lipid deposition, and fibrosis were evaluated. The results showed that none of the mice died during the treatment with Linba Fang. The levels of tail swelling, inflammation, lipid deposition, and fibrosis in mice treated with Linba Fang were significantly decreased compared with negative and positive controls. Among mice treated with the same dose of Linba Fang, the levels of tail swelling, inflammation, lipid deposition, and fibrosis in mice treated for 4 weeks were significantly lower than those treated for 2 weeks. Among mice treated for the same duration of time, the levels of tail swelling, inflammation, lipid deposition, and fibrosis showed a decreasing tendency following increasing doses. Notably, the inflammation in tail tissues decreased to the similar level of normal group after treatment for 4 weeks using the high dose of Linba Fang. In conclusion, the traditional Chinese medical prescription Linba Fang could inhibit the pathological changes caused by lymphedema, including swelling, inflammation, lipid deposition, and fibrosis.

Determination of the Functional Membrane Integrity of Mice Spermatozoa Using the Classic Hypoosmotic Swelling Test (HOST) and Modified Host

The hypo-osmotic swelling test (HOST) is widely used in determining sperm quality by evaluating membrane integrity in both human and animal spermatozoa. The test measures the swelling ability and membrane integrity of functional sperm in hypo-osmotic solutions. The ability of the sperm membrane to maintain equilibrium between itself and the environment indicates a functional and intact plasma membrane. This functionality is depicted by the coiling and ballooning or "swelling” of the sperm tail when influx of fluid occurs due to hypo-osmotic stress. The modified HOST solution consists of Earle's Buffer Salt Solution (EBSS) while the classic HOST solution consists of sodium citrate and fructose. It has been reported that modified HOST solution show more rapid sperm tail swelling and better viability in human sperm cells. This study aims to compare the swelling ability and viability of mouse sperm upon incubation in classic HOST medium and modified HOST medium. Iso-osmotic solution was used as negative control, while distilled water acts as positive control. Caudal epididymal sperm from twenty mature male mice were collected into Kreb Ringer's Buffer medium and divided into 4 batches, one for each test solution. Sperm were pooled and discarded of cell debris and centrifuged in a discontinuous Ficoll gradient (45% and 90%). Sperm were collected at the pellet and incubated for 60 minutes in the respective test solutions. Tail swelling and viability were observed from smears taken at intervals of 5 minutes. ANOVA analysis showed no significant difference between the classic HOST and modified HOST in sperm ‘ballooning' and sperm viability. The results suggests that using modified HOST showed no significant improvement from classic HOST in functional membrane integrity changes in mouse spermatozoa as compared to human spermatozoa.

Molecular aspect of silver nanoparticles regulated embryonic development in Zebrafish (Danio rerio) by Oct-4 expression

With the enhancement of commercial manifestation of silver nanoparticles, concerned has risen on their accumulation in aquatic system and consequent effects on fish development and metabolism. In this study, experiments were conducted to assess the impacts of silver nanoparticles on early life cycles of fish considering Zebrafish (Danio rerio) as experimental model. Silver nanoparticles were synthesized through chemical reduction method and characterized through UV–visible spectroscopy, dynamic light scattering (DLS), and HR-TEM. Different sub lethal doses of nanosilver were applied (13.6, 21.6, 42.4, 64, and 128 μgL−1) to post-fertilization phases of Zebrafish embryos and their interaction effects were monitored up to six days period. No significant morphological variations were observed at 13.6, 21.6, 42.4 μgL−1 dose of silver nanoparticles, whereas 64 and 128 μgL−1 exposure dose exhibited bending in myotome, deformity in tail region, somites, notochord and swelling in anterior and posterior region of embryos and larva. Hatching performances analysis elicited highest hatching success in 13.6 and 21.6 μgL−1 doses of silver nanoparticles followed by positive and negative control, whereas exposure dose of 64 and 128 μgL−1 exhibited comparatively lower success. Western blot analysis were conducted on developing hatchlings with Oct4 antibody and at 13.6 and 21.6 μgL−1dose,it showed over expression elucidating stimulatory role of nanosilver in these mentioned doses. In silico analysis depicted a firm interaction of nanosilver with Oct4 revealing their key role in growth stimulation of developing embryos. The study demonstrates the function of nanosilver as a growth promoter rather only as a toxicant in fish metabolic system.

Sperm with an abnormal hypo-osmotic swelling test – normal fertilization, normal embryo development, but implantation failure

Purpose: To review the clinical importance of including the hypo-osmotic swelling (HOS) test in routine male fertility testing which in general is not evaluated by most physicians dealing with infertility. Materials and Methods: Pregnancy rates were evaluated in patients with low HOS test scores. A low HOS test was specifically defined as having less than 50% of sperm exhibiting the normal physiologic response of tail swelling, when subjected to a hypo-osmolar solution. Pregnancy rates of patients with low HOS test were examined after intercourse, intrauterine insemination (IUI), conventional oocyte insemination, and in vitro fertilization (IVF). Patients with a low HOS test were also treated with a protein digestive enzyme chymotrypsin. Patients receiving intervention then underwent IUI, IVF with conventional oocyte insemination, or IVF with intracytoplasmic sperm injection (ICSI). Pregnancy rates of the cohort receiving intervention were then examined for comparison. Results: The HOS test abnormality leads to normal fertilization but almost invariably negatively effects embryo implantation. Treatment with chymotrypsin, or performing IVF with ICSI, can overcome the toxic protein causing the embryo implantation defect. This toxic protein may be cryolabile and freezing sperm or embryos may prove to be another mode of therapy. Conclusions: The HOS abnormality may be the most reliable semen abnormality predicting failure to conceive even with IVF unless the defect is negated. Therapy is very effective. Unfortunately this test is rarely evaluated by most infertility specialists but it should be. The frequency increases with age.

Alaria mesocercariae in the tails of red-sided garter snakes: evidence for parasite-mediated caudectomy.

Trematodes of the genus Alaria develop into an arrested stage, known as mesocercariae, within their amphibian second intermediate host. The mesocercariae are frequently transmitted to a non-obligate paratenic host before reaching a definitive host where further development and reproduction can occur. Snakes are common paratenic hosts for Alaria spp. with the mesocercariae often aggregating in the host's tail. In the current study, we used morphological examination and molecular analyses based on partial sequences of nuclear large ribosomal subunit gene and mitochondrial cytochrome C oxidase subunit 1 gene to identify larvae in the tails of red-sided garter snakes (Thamnophis sirtalis parietalis) as mesocercariae of Alaria marcianae, Alaria mustelae, and Alaria sp. as well as metacercariae of Diplostomidae sp. of unknown generic affiliation. We assessed infection prevalence, absolute and relative intensity, and associated pathological changes in these snakes. Infection prevalence was 100% for both male and female snakes. Infection intensity ranged from 11 to more than 2000 mesocercariae per snake tail but did not differ between the sexes. Gross pathological changes included tail swelling while histopathological changes included mild inflammation and the presence of mucus-filled pseudocysts surrounding mesocercariae, as well as the compression and degeneration of muscle fibers. Our results indicate that mesocercariae can lead to extensive muscle damage and loss in both sexes which likely increases the fragility of the tail making it more prone to breakage. As tail loss in garter snakes can affect both survival and reproduction, infection by Alaria mesocercariae clearly has serious fitness implications for these snakes.

Cimetidine attenuates vinorelbine-induced phlebitis in mice by militating E-selectin expression

We investigated E-selectin expression in mice and rabbits with vinorelbine-induced phlebitis and the effect of cimetidine. To find the relationship between E-selectin expression and vinorelbine-induced phlebitis. Mouse and rabbit model of vinorelbine-induced phlebitis was established by intravenous infusion of vinorelbine. Pathological observation, molecular-biological determination of E-selectin and protein function of it was evaluated. Grossly, we observed swelling, edema and cord-like vessel changes in mice receiving vinorelbine but only mild edema in mice pretreated with cimetidine. Pathological scoring yielded a total score of 37 for vinorelbine-treated mice and 17 for mice pretreated with cimetidine (P < 0.05). ELISA revealed that rabbits treated with vinorelbine had markedly higher serum contents of E-selectin than normal saline (NS) controls (vinorelbine 1.534 ± 0.449 vs. NS 0.746 ± 0.170 ng/mL, P < 0.05), which was markedly attenuated by cimetidine (cimetidine 0.717 ± 0.468 vs. vinorelbine 1.534 ± 0.449 ng/mL, P < 0.05). Rose Bengal staining assays showed that vinorelbine markedly increased the adhesion rate of neutrophils for endothelial cells (vinorelbine 38.70 ± 8.34% vs. controls 8.93 ± 4.85%, P < 0.01), which, however, was significantly suppressed by cimetidine (9.93 ± 5.91%, P < 0.01 vs. vinorelbine). In E-selectin knockout mice, we found no apparent difference in tail swelling in mice receiving vinorelbine or cimetidine and vinorelbine. In conclusion, cimetidine attenuates vinorelbine-induced phlebitis in mice probably by suppressing increased expression of E-selectin.

The Multicomponent Medication Lymphomyosot Improves the Outcome of Experimental Lymphedema

Secondary lymphedema is a life-long disease of painful tissue swelling that often follows axillary lymph node dissection to treat breast cancer. It is hypothesized that poor lymphatic regeneration across the obstructive scar tissue during the wound healing process may predispose the tissue to swell at a later date. Treatment for lymphedema remains suboptimal and is in most cases palliative. The purpose of this study was to evaluate the ability of Lymphomyosot to treat tissue swelling and promote lymphangiogenesis in experimental models of murine lymphedema. Experimental models of mouse lymphedema were injected with varied amounts of Lymphomyosot and saline as control. Measurements of tail swelling and wound closure were taken and compared amongst the groups. Three separate groups of mice were analyzed for lymphatic capillary migration, lymphatic vessel regeneration, and macrophage recruitment. Lymphomyosot significantly reduced swelling and increased the rate of surgical wound closure. Lymphomyosot did not increase the migration of lymph capillaries in a mouse tail skin regeneration model or regeneration of lymph vessels following murine axillary lymph node dissection. Lymphomyosot may act through inflammatory and wound repair pathways to reduce experimental lymphedema. Its ability to regulate inflammation as well as assist in tissue repair and extracellular formation may allow for the production of a scar-free matrix bridge through which migrating cells and accumulated interstitial fluid can freely spread.

Evaluation of subgroups of the human sperm hypoosmotic swelling test in normozoospermic male cases with recurrent fertilization failure: a prospective case-controlled study

To compare subgroups of the human sperm hypoosmotic swelling test subgroups in both recurrent fertilization negative infertile cases with normal semen analysis and fertilization positive controls. This was a prospective case-controlled study performed with normospermic 33 previously fertile male (secondary infertility) and 41 infertile men who had undergone two or three unsuccessful in vitro fertilization attempts. HOS test was investigated in 4 subgroups including HOS 1, HOS 2, HOS 3, and HOS 4 according to the degree of sperm tail swelling and compared between the two groups. Four subgroups were compared and statistical significance was demonstrated in HOS 1, HOS 3 and HOS 4 tests (p < 0.001) in fertile and infertile men. Highest HOS 1 and lowest HOS 4 grades were determined in Group A. However, no statistical significance was determined between two groups in HOS 2 test which was minimal swelling in sperm tails. HOS 1, HOS 3 and HOS 4 subgroups of HOS test are reliable and useful methods providing important information regarding the sperm function. A high HOS test 1 grade plus a low HOS test 4 grade should suggest a fertility problem, despite a normal semen analysis. HOS test subgroups provide additional information in normospermic cases with unexplained infertility.

Increased Hyaluronan Expression at Distinct Time Points in Acute Lymphedema

Lymphatic dysfunction in lymphedema results in chronic accumulation of interstitial fluid and life-long tissue swelling. In the absence of restored lymphatic drainage via adequate lymphangiogenesis, the interstitial environment can remodel in ways that decrease the elevated interstitial stress. Presently, relatively little is known about the glycosaminoglycans (GAGs) that become upregulated in the interstitium during lymphedema. We employed a mouse tail model of acute lymphedema that reproduces important features of the chronic human condition to establish a relationship between hyaluronan (HA) and sulfated GAG concentration with tissue swelling. We found that HA was upregulated by tissue injury at day 5 and became upregulated again by skin swelling (HA content increasing by 27% relative to controls at days 15 and 20). Surprisingly, the second phase of HA expression was associated with the declining phase of the tail skin swelling (tail diameter significantly decreasing by 17% from day 10 peak to day 20), demonstrating that HA is upregulated by tissue swelling and may help to counteract the edema in the mouse tail. This finding was confirmed by intradermal injection of an HA degrading enzyme (hyaluronidase) to the swollen tail, which was found to worsen the tail swelling. Sulfated GAGs, including chondroitin sulfate (CS), were not regulated by tissue swelling. The results demonstrate that HA, but not sulfated GAGs, is upregulated in the interstitium by acute tissue swelling. We speculate that HA expression during lymphedema may be part of a natural adaptive mechanism of the interstitial environment to reduce capillary filtration and increase interstitial fluid outflow following lymphatic obstruction and fluid accumulation.

Pancreatitis with Pancreatic Tail Swelling Associated with Incretin-based Therapies Detected Radiologically in Two Cases of Diabetic Patients with End-Stage Renal Disease

We herein report two cases of pancreatitis associated with incretin-based therapies in end-stage renal disease (ESRD) patients undergoing dialysis. A 75-year-old woman with a history of liraglutide use and a 68-year-old man with a history of vildagliptin use both presented with nausea. They showed elevated levels of pancreatic enzymes and pancreatic tail swelling on CT. Their symptoms improved after discontinuing the drugs. In the absence of any obvious secondary causes of pancreatitis, we believe that the pancreatitis observed in these cases was associated with the incretin-based therapies. Few reports have been published on the safety and efficacy of incretin-based therapies in ESRD patients, and it remains uncertain whether the changes in the pancreas observed in the present cases are characteristic of ESRD patients.

Mechanism for sperm immobilization activity of extract from Platycodon grandiflorum in vitro

To explore the mechanism of spermicidal effect of crude extract and platycodin-D from Platycodon grandiflorum (PG) root in vitro. Between February 2006 and December 2009, 38 fertile and healthy adult males were selected as donors. PG root was extracted and platycodin-D purified. Grouping was as follows: crude extract from PG root, platycodin-D, nonoxynol-9 (N-9, as a reference standard) and semen-added physiological saline (as control). Spermicidal experiments were carried out in vitro (Sander-Cramer test). The hypo-osmotic swelling (HOS) test and modified Eosin-Giemsa (EG) staining were used to detect the integrity of sperm membrane. Four types of sperm morphology were divided through HOS-EG test: Type A: spermatozoa with swelling in tails and head white staining HOS(+)-EG(-) (membrane intact); Type B: spermatozoa with no swelling in tails (membrane-damaged) and head white staining HOS(-)-EG(-); Type C: spermatozoa with tail swelling and head red HOS(+)-EG(+); Type D: spermatozoa with no swelling in tails and head red HOS(-)-EG(+). Sperm chromatin dispersion (SCD) test was performed to determine the integrity of sperm DNA. The crude extract from PG root could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 50.0 g/L and 20.0 g/L (v:v = 1:1 in semen). When the semen sample was exposed to the concentrations of 2.0 g/L and 1.0 g/L of platycodin-D, all spermatozoa were immobilized within 20 s. In the control group, the mean percentage of Types A, B, C and D was (69.0 ± 8.3)%, (3.4 ± 0.5)%, (10.2 ± 1.7)% and (17.4 ± 2.1)% respectively. In the groups of platycodin-D and N-9 solution, the rate of Types A and B was 0. The rate of Types C [(65.3 ± 3.8)%] and D [(34.7 ± 7.1)%] significantly increased versus control in the platycodin-D group (P < 0.01). Sperm DNA fragmentation had no change upon an exposure to the extract from PG root, platycodin-D and N-9 solution. And the sperm revival test showed none of the spermatozoa recovered their motility. The extract and platycodin-D from PG root have a quick sperm-killing effect in a short time in vitro by disrupting the integrity of sperm membrane (main head).