Asymmetry in kinematics of dominant/nondominant lower limbs in central and lateral positioned college and sub-elite soccer players.

Change of direction, stops, and pivots are among the most common non-contact movements associated with anterior cruciate ligament (ACL) injuries in soccer. By observing these dynamic movements, clinicians recognize abnormal kinematic patterns that contribute to ACL tears such as increased knee valgus or reduced knee flexion. Different motions and physical demands are observed across playing positions, which may result in varied lower limb kinematic patterns. In the present study, 28 college and sub-elite soccer players performed four dynamic motions (change of direction with and without ball, header, and instep kick) with the goal of examining the effect of on-field positioning, leg dominance, and gender in lower body kinematics. Motion capture software monitored joint angles in the knee, hip, and ankle. A three-way ANOVA showed significant differences in each category. Remarkably, centrally positioned players displayed significantly greater knee adduction (5° difference, p = 0.013), hip flexion (9° difference, p = 0.034), hip adduction (7° difference, p = 0.016), and dorsiflexion (12° difference, p = 0.022) when performing the instep kick in comparison to their laterally positioned counterparts. These findings suggest that central players tend to exhibit a greater range of motion when performing an instep kicking task compared to laterally positioned players. At a competitive level, this discrepancy could potentially lead to differences in lower limb muscle development among on-field positions. Accordingly, it is suggested to implement position-specific prevention programs to address these asymmetries in lower limb kinematics, which can help mitigate dangerous kinematic patterns and consequently reduce the risk of ACL injury in soccer players.

MIF-Modulated Spinal Proteins Associated with Persistent Bladder Pain: A Proteomics Study.

Bladder pain is a prominent symptom in Interstitial Cystitis/Bladder Pain Syndrome (IC/BPS). We studied spinal mechanisms of bladder pain in mice using a model where repeated activation of intravesical Protease Activated Receptor-4 (PAR4) results in persistent bladder hyperalgesia (BHA) with little or no bladder inflammation. Persistent BHA is mediated by spinal macrophage migration inhibitory factor (MIF), and is associated with changes in lumbosacral proteomics. We investigated the contribution of individual spinal MIF receptors to persistent bladder pain as well as the spinal proteomics changes associated with relief of persistent BHA by spinal MIF antagonism. Female mice with persistent BHA received either intrathecal (i.t.) MIF monoclonal antibodies (mAb) or mouse IgG1 (isotype control antibody). MIF antagonism temporarily reversed persistent BHA (peak effect: 2 h), while control IgG1 had no effect. Moreover, i.t. antagonism of the MIF receptors CD74 and C-X-C chemokine receptor type 4 (CXCR4) partially reversed persistent BHA. For proteomics experiments, four separate groups of mice received either repeated intravesical scrambled peptide and sham i.t. injection (control, no pain group) or repeated intravesical PAR4 and: sham i.t.; isotype IgG1 i.t. (15 µg); or MIF mAb (15 µg). L6-S1 spinal segments were excised 2 h post-injection and examined for proteomics changes using LC-MS/MS. Unbiased proteomics analysis identified and relatively quantified 6739 proteins. We selected proteins that showed significant changes compared to control (no pain group) after intravesical PAR4 (sham or IgG i.t. treatment) and showed no significant change after i.t. MIF antagonism. Six proteins decreased during persistent BHA (V-set transmembrane domain-containing protein 2-like confirmed by immunohistochemistry), while two proteins increased. Spinal MIF antagonism reversed protein changes. Therefore, spinal MIF and MIF receptors mediate persistent BHA and changes in specific spinal proteins. These novel MIF-modulated spinal proteins represent possible new targets to disrupt spinal mechanisms that mediate persistent bladder pain.

Lumbosacral spinal proteomic changes during PAR4-induced persistent bladder pain.

Repeated intravesical activation of protease-activated receptor-4 (PAR4) in mice results in persistent bladder hyperalgesia (BHA). We investigated spinal proteomic changes associated with persistent BHA. Persistent BHA was induced in female mice by repeated (3x; days 0,2,4; n = 9) intravesical instillation of PAR4 activating peptide (PAR4-AP) while scrambled peptide served as the control (no pain; n = 9) group. The threshold to lower abdominal von Frey stimulation was recorded prior to and during treatment. On day 7, L6-S1 spinal segments were excised and examined for proteomic changes using LC-MS/MS. In-depth, unbiased proteomic tandem-mass tag (TMT) analysis identified and relatively quantified 6739 proteins. We identified significant changes with 29 decreasing and 51 increasing proteins in the persistent BHA group and they were associated with neuroprotection, redox modulation, mitochondrial factors, and neuronal-related proteins. In an additional experiment, decreases in protein levels were confirmed by immunohistochemistry for metallothionein 1/2. Our results show that persistent bladder pain is associated with central (spinal) protein changes. Previous work showed that PAR4-induced bladder pain is mediated, at least in part by spinal MIF. Further functional studies of these top changing proteins may lead to the discovery of novel potential therapeutic targets at the spinal level to modulate persistent bladder pain. Future studies will examine the effect of spinal MIF antagonism on PAR4-induced spinal proteomics associated with persistent bladder pain.

Urothelial Oxidative Stress and ERK Activation Mediate HMGB1-Induced Bladder Pain.

Activation of intravesical protease activated receptors-4 (PAR4) results in bladder pain through the release of urothelial macrophage migration inhibitory factor (MIF) and high mobility group box-1 (HMGB1). We aimed to identify HMGB1 downstream signaling events at the bladder that mediate HMGB1-induced bladder pain in MIF-deficient mice to exclude any MIF-related effects. We studied whether oxidative stress and ERK activation are involved by examining bladder tissue in mice treated with intravesical disulfide HMGB1 for 1 h and analyzed with Western blot and immunohistochemistry. HMGB1 intravesical treatment increased urothelium 4HNE and phospho-ERK1/2 staining, suggesting that HMGB1 increased urothelial oxidative stress and ERK activation. Furthermore, we examined the functional roles of these events. We evaluated lower abdominal mechanical thresholds (an index of bladder pain) before and 24 h after intravesical PAR4 or disulfide HMGB1. Intravesical pre-treatments (10 min prior) included: N-acetylcysteine amide (NACA, reactive oxygen species scavenger) and FR180204 (FR, selective ERK1/2 inhibitor). Awake micturition parameters (voided volume; frequency) were assessed at 24 h after treatment. Bladders were collected for histology at the end of the experiment. Pre-treatment with NACA or FR significantly prevented HMGB1-induced bladder pain. No significant effects were noted on micturition volume, frequency, inflammation, or edema. Thus, HMGB1 activates downstream urothelial oxidative stress production and ERK1/2 activation to mediate bladder pain. Further dissection of HMGB1 downstream signaling pathway may lead to novel potential therapeutic strategies to treat bladder pain.

Fisioterapia y osteoartritis: una revisión histórica de las técnicas utilizadas en el pasado y su relevancia actual / Physiotherapy and osteoarthritis: a historical review of techniques used in the past and their current relevance

Objetivo principal: El propósito de esta revisión ha sido proveer una visión histórica de las técnicas de fisioterapia empleadas en el pasado y su relevancia actual en el tratamiento de la Osteoartritis (OA). Metodología: Se realizó una búsqueda exhaustiva en bases de datos y análisis de la literatura. Se incluyeron artículos que abordaran el tratamiento de la OA en fisioterapia desde la Historia Antigua hasta la actualidad. Resultados principales: Se seleccionaron 34 publicaciones que abordaron el tratamiento de la OA en diferentes períodos históricos. Las técnicas identificadas fueron desde tratamientos a base de ungüentos hasta enfoques actuales como la hidroterapia y la electroterapia. Conclusión principal: El abordaje fisioterapéutico de la OA ha evolucionado a lo largo de la historia con el empleo de diversas técnicas y agentes físicos. Desde el siglo XIX las técnicas empleadas lograron un avance científico y terapéutico valioso para esta afección (AU)

Objective: The purpose of this review has been to provide a historical perspective on the physiotherapy techniques used in the past and their current relevance in the treatment of Osteoarthritis (OA). Methods: A comprehensive search of databases and literature analysis was conducted. Articles addressing the treatment of OA in physiotherapy from Ancient History to the present were included. Results: 34 publications addressing the treatment of OA in different historical periods were selected. The identified techniques ranged from ointment-based treatments to current approaches such as hydrotherapy and electrotherapy. Conclusions: The physiotherapeutic approach to OA has evolved throughout history, utilizing various techniques and physical agents. Since the 19th century, the employed techniques have achieved valuable scientific and therapeutic advancements for this condition (AU)

Fisioterapia y osteoartritis: una revisión histórica de las técnicas utilizadas en el pasado y su relevancia actual / Physiotherapy and osteoarthritis: a historical review of techniques used in the past and their current relevance

Objetivo principal: El propósito de esta revisión ha sido proveer una visión histórica de las técnicas de fisioterapia empleadas en el pasado y su relevancia actual en el tratamiento de la Osteoartritis (OA). Metodología: Se realizó una búsqueda exhaustiva en bases de datos y análisis de la literatura. Se incluyeron artículos que abordaran el tratamiento de la OA en fisioterapia desde la Historia Antigua hasta la actualidad. Resultados principales: Se seleccionaron 34 publicaciones que abordaron el tratamiento de la OA en diferentes períodos históricos. Las técnicas identificadas fueron desde tratamientos a base de ungüentos hasta enfoques actuales como la hidroterapia y la electroterapia. Conclusión principal: El abordaje fisioterapéutico de la OA ha evolucionado a lo largo de la historia con el empleo de diversas técnicas y agentes físicos. Desde el siglo XIX las técnicas empleadas lograron un avance científico y terapéutico valioso para esta afección. (AU)

Objective: The purpose of this review has been to provide a historical perspective on the physiotherapy techniques used in the past and their current relevance in the treatment of Osteoarthritis (OA). Methods: A comprehensive search of databases and literature analysis was conducted. Articles addressing the treatment of OA in physiotherapy from Ancient History to the present were included. Results: 34 publications addressing the treatment of OA in different historical periods were selected. The identified techniques ranged from ointment-based treatments to current approaches such as hydrotherapy and electrotherapy. Conclusions: The physiotherapeutic approach to OA has evolved throughout history, utilizing various techniques and physical agents. Since the 19th century, the employed techniques have achieved valuable scientific and therapeutic advancements for this condition. (AU)

Bladder Oxidative Stress and HMGB1 Release Contribute to PAR4-Mediated Bladder Pain in Mice.

Activation of intravesical PAR4 receptors leads to bladder hyperalgesia (BHA) through release of urothelial macrophage migration inhibitory factor (MIF) and urothelial high mobility group box-1 (HMGB1). MIF deficiency and/or MIF antagonism at the bladder block BHA in mice yet the mechanisms are not clear. Since oxidative stress and ERK phosphorylation are involved in MIF signaling we hypothesized that oxidative stress and/or ERK signaling, activated by MIF release, promote intravesical HMGB1 release to induce BHA. We induced BHA by intravesical PAR4 infusion in female C57BL/6 mice. Mechanical sensitivity was evaluated by measuring abdominal von Frey (VF) 50% thresholds before (baseline) and 24 h post-infusion. Intravesical pre-treatment (10 min infusion prior to PAR4) with N-acetylcysteine amide (NACA; reactive-oxygen species scavenger; 3 mg in 50 µl), FR180204 (selective ERK1/2 inhibitor; 200 µg in 50 µl), ethyl pyruvate (EP; HMGB1 release inhibitor; 600 µg in 50 µl), or diluent controls (50 µl) tested the effects of pre-treatment on PAR4-induced BHA. Intravesical fluid was collected after each treatment and HMGB1 concentration was measured using ELISA. Awake micturition parameters (volume and frequency) were assessed at the end of the experiments. Bladders were collected and examined for histological signs of edema and inflammation. Pre-treatment with PBS followed by PAR4 induced BHA in mice but PBS followed by scrambled peptide did not. Pre-treatment with NACA or EP partially blocked PAR4-induced BHA while FR180204 had no effect. A significant correlation between intravesical HMGB1 levels and 50% VF thresholds was observed. All PAR4 treated groups had increased levels of HMGB1 in the intravesical fluid compared to PBS-Scrambled group although not statistically significant. No significant effects were noted on awake micturition volume, micturition frequency or histological evidence of bladder edema or inflammation. Our results show that intravesical antagonism of bladder reactive-oxygen species accumulation was effective in reducing PAR4-induced bladder pain. The correlation between intravesical levels of HMGB1 and bladder pain indicates that released HMGB1 is pivotal to bladder pain. Thus, modulating events in the MIF signaling cascade triggered by PAR4 activation (including bladder oxidative stress and HMGB1 release) warrant further investigation as possible therapeutic strategies.

Intravesical CD74 and CXCR4, macrophage migration inhibitory factor (MIF) receptors, mediate bladder pain.

BACKGROUND: Activation of intravesical protease activated receptor 4 (PAR4) leads to release of urothelial macrophage migration inhibitory factor (MIF). MIF then binds to urothelial MIF receptors to release urothelial high mobility group box-1 (HMGB1) and elicit bladder hyperalgesia. Since MIF binds to multiple receptors, we investigated the contribution of individual urothelial MIF receptors to PAR4-induced HMGB1 release in vivo and in vitro and bladder pain in vivo. METHODOLOGY/PRINCIPAL FINDINGS: We tested the effect of intravesical pre-treatment with individual MIF or MIF receptor (CD74, CXCR4, CXCR2) antagonists on PAR4-induced HMGB1 release in vivo (female C57/BL6 mice) and in vitro (primary human urothelial cells) and on PAR4-induced bladder hyperalgesia in vivo (mice). In mice, PAR4 induced HMGB1 release and bladder hyperalgesia through activation of intravesical MIF receptors, CD74 and CXCR4. CXCR2 was not involved in these effects. In primary urothelial cells, PAR4-induced HMGB1 release through activation of CD74 receptors. Micturition parameters in mice were not changed by any of the treatments. CONCLUSIONS/SIGNIFICANCE: Urothelial MIF receptors CD74 and CXCR4 mediate bladder pain through release of urothelial HMGB1. This mechanism may set up persistent pain loops in the bladder and warrants further investigation. Urothelial CD74 and CXCR4 may provide novel targets for interrupting bladder pain.

Beauveria bassiana Multifunction as an Endophyte: Growth Promotion and Biologic Control of Trialeurodes vaporariorum, (Westwood) (Hemiptera: Aleyrodidae) in Tomato.

The tomato, Solanum lycopersicum L. is one of the most consumed vegetables in the world; nevertheless, it is affected by biotic and abiotic factors that reduce its productivity. The whitefly is globally considered as the main pest under protected crop conditions, where biologic control using endophytic fungi emerges as a sustainable alternative. We evaluated the indirect effects of five native endophytic strains of Beauveria bassiana on the reproduction of greenhouse whiteflies and the growth of tomatoes. The plant growth substrate was inoculated with five strains of this endophyte and the resulting plants were then exposed to whiteflies afterwards. The effect that endophytic strains had on phosphate solubilization, iron siderophore production, plant height, and plant biomass were evaluated. The evaluated endophytes reduced the number of eggs per cm2 on leaflets compared to the control and behaved similarly to the commercial synthetic insecticide. Leaflets inoculated with strains RGM-557, RGM-644 and RGM-731 showed fewer nymphs than the control and those treated with insecticide. RGM-557 and RGM-731 produced the greatest plant heights; RGM-731 obtained the greatest plant biomass. Our study provides evidence that native endophytic strains of B. bassiana have a biocontrol effect on whiteflies and could be used to promote tomato growth.

Anatomía topográfica del nervio laríngeo superior: importancia quirúrgica en las tiroidectomías / Topographic anatomy of the upper laryngeal nerve: surgical importance in thyroidectomies

En las cirugías sobre la glándula tiroides se ha prestado mucha atención al manejo del nervio laríngeo inferior y de las glándulas paratiroides, no así del ramo lateral del nervio laríngeo superior, el cual es satélite del pedículo vascular superior de la glándula tiroides. El manejo del polo superior de la glándula tiroides requiere de un conocimiento acabado de su anatomía topográfica del área y está determinada por sus relaciones más importantes, dadas por el citado nervio y la arteria tiroidea superior principalmente. En este trabajo se pretende estudiar estas relaciones en base a la disección meticulosa del triángulo laringo-esternotiroideo ("Triángulo de Joll") de 25 cadáveres adultos formolizados. Como hallazgo relevante se informa que los "nervios en riesgo", según la clasificación de Cernea, que se basa en una distancia menor a un centímetro en el entrecruzamiento del nervio con la arteria tiroidea superior con respecto al polo superior de la glándula tiroides, es del 52 % para el lado derecho y 44 % para el lado izquierdo del cuello. El origen bajo de la arteria a nivel de la bifurcación carotídea se presenta asociada a un mayor número de "nervios en riesgo" en el lado izquierdo. Según el punto de penetración del ramo lateral del nervio laríngeo superior en el músculo constrictor inferior de la faringe se establece la clasificación de Friedman, muy útil sobre todo en cirugías ayudadas por la neuroestimulación. En esta clasifiacción los "nervios en riesgo" son aquellos que transcurrren superficial al músculo, mientras que los "nervios protegidos" serían aquellos que perforan el músculo en su porción superior. En este trabajo los "nervios en riesgo" se presentaron del lado izquierdo en el 56 % de los casos y del derecho en el 60 %, mientras que los "nervios protegidos" en el 24 % y 16 %, respectivamente.

In surgeries on the thyroid gland, much attention has been given to the management of the inferior laryngeal nerve and parathyroid glands, but not the external branch of the given by the aforementioned nerve and the superior thyroid artery. This paper intends to study these relationships based on the meticulous dissection of the larynx-sternothyroid triangle ("Joll triangle") of 25 formolized adult corpses. As a relevant finding, it is reported that the " nerves at risk" according to the Cernea classification, which is based on the distance less than one centimeter at the intersection of the nerve with the superior thyroid artery with respect to the upper pole of the gland, is 52 % for the right side and 44 % for the left side of the neck. The low origin of the artery at the level of the carotid bifurcation is associated with a greater number of "nerves at risk" on the left side. According to the penetration point of the external branch of the superior laryngeal nerve in the inferior pharyngeal constrictor muscle, the Friedman classification is established, very useful especially in surgeries aided by neurostimulation. In this classification the "nerves at risk" are those that run superficially to the muscle, while the protected nerves would be those that pierce the muscle in its upper part. In tis work, the "nerves at risk" presented on the left side in 56 % of the cases and the right side in 60 %, while those "protected" in 24 % and 16 % respectively.

MIF mediates bladder pain, not inflammation, in cyclophosphamide cystitis.

Macrophage migration inhibitory factor (MIF), a proinflammatory mediator, is recognized as a player in inflammatory and neuropathic pain. Cyclophosphamide (CYP) results in bladder inflammation and pain and it's a frequently used animal model of interstitial cystitis/bladder pain syndrome (IC/BPS). Because pretreatment with a MIF inhibitor (ISO-1) prevented both CYP-induced bladder pain and inflammation we used genetic MIF knockout (KO) mice to further investigate MIF's role in CYP-induced bladder pain and inflammation. Abdominal mechanical threshold measured bladder pain induced by CYP in wild type (WT) and MIF KO mice at several time points (0-48 hours). End-point (48 hours) changes in micturition parameters and histological signs of bladder inflammation were also evaluated. Abdominal mechanical hypersensitivity developed within 4 hours after CYP injection (and lasted for the entire observation period: 48 hours) in WT mice. MIF KO mice, on the other hand, did not develop abdominal mechanical hypersensitivity suggesting that MIF is a pivotal molecule in mediating CYP-induced bladder pain. Both WT and MIF KO mice treated with CYP showed histological signs of marked bladder inflammation and showed a significant decrease in micturition volume and increase in frequency. Since both changes were blocked in MIF KO mice by pretreatment with a MIF inhibitor (ISO-1) it is likely these are non-specific effects of ISO-1. MIF mediates CYP-induced bladder pain but not CYP-induced bladder inflammation. The locus of effect (bladder) or central (spinal) for MIF mediation of bladder pain remains to be determined.

Erectile Dysfunction Medications and Skin Cancer: An Analysis in US Veterans.

OBJECTIVE: To examine the relationship between phosphodiesterase 5 inhibitor drugs (PDE5i) and skin cancers in a large-scale study of Veterans. METHODS: This was a retrospective database review using the Department of Veterans Affairs Informatics and Computing Infrastructure database. Veterans Affairs Informatics and Computing Infrastructure was searched 19 years for Veterans who received PDE5i treatment of erectile dysfunction. A non-PDE5i group of Veterans was selected for comparison analysis. Follow-up time, outpatient clinic visits and incidence of malignant melanoma (MM), and basal cell carcinoma (BCC) were measured in both groups. RESULTS: A total of 2.55 million Veterans were included in this study (1.27 million in each group). Mean age of the PDE5i group and non-PDE5i group was 59.2 years (standard deviation [SD] ± 10.8) and 58.7 (SD ± 10.8), respectively. Mean follow-up time for the PDE5i group was 8.9 years (SD ± 4.2) and 8.5 years (SD ± 4.3) for non-PDE5i group. Odds ratio for malignant melanoma and BCC in the PDE5i group was 1.25 (confidence interval 95%, 1.22-1.28, P <.0001) and 1.49 (confidence interval 95%, 1.46-1.51, P <.0001), respectively. PDE5i users showed more mean outpatient visits/year (8.9 SD ± 9.50) compared to non-PDE5i users (5.9 SD ± 10.0; P <.0001). CONCLUSION: Veterans prescribed PDE5is to treat erectile show a minimal increased risk of MM and a greater risk of BCC compared to non-PDE5i users. PDE5i users visited outpatient VA clinics at a higher rate than non-PDE5i users in this study. These findings suggest confounding variables are likely involved in the relationship between skin cancers and PDE5i use. PDE5i drugs remain a safe treatment for erectile dysfunction.

Spinal macrophage migration inhibitory factor and high mobility group box 1 mediate persistent bladder pain.

Repeated intravesical PAR4 (protease activated receptor 4) activation elicits persistent bladder pain lasting 5 days after the last treatment. Persistent bladder pain was fully reversed by a systemic HMGB1 (high mobility group box 1) inhibitor while a MIF (macrophage migration inhibitory factor) antagonist partly reversed it. Since there is growing evidence that spinal MIF and HMGB1 mediate inflammatory and neuropathic pain we examined whether there were spinal changes occurring during persistent bladder pain that may be responsible for maintaining bladder pain. In addition, we tested whether we could modulate persistent bladder pain with spinal MIF or HMGB1 antagonists. Persistent bladder pain was elicited in female C57 mice by repeated (3x) intravesical instillation of PAR4-activating peptide while control animals received scramble peptide treatment. On day 4, spinal cord (L6-S1) changes in c-fos (non-specific marker of spinal activation) was assessed with immunofluorescence while MIF and HMGB1 were assessed with immunofluorescence, western blotting and real-time PCR. On day 7, mice received an intrathecal injection of a neutralizing MIF monoclonal antibody (15 µg in 5 µl PBS) or a HMGB1 inhibitor glycyrrhizin (25 µg in 5 µl of 5% alcohol in PBS) and abdominal mechanical threshold was tested. On day 9, mice were treated with vehicle or control and abdominal mechanical threshold was tested. Immunofluorescence showed that c-fos and MIF in the dorsal horn, dorsal grey commissure and intermediolateral areas significantly increased in PAR4-treated mice while HMGB1 was decreased. In addition, intrathecal treatment with MIF neutralizing mAb or glycyrrhizin significantly alleviated abdominal mechanical hypersensitivity at 1 and 2 h and the analgesic effect diminished at 6 h. Vehicle or control treatment had no effect. Persistent bladder pain is associated with spinal changes in MIF and HMGB1 levels. Furthermore, spinal treatment with MIF monoclonal antibody and HMGB1 inhibitor temporarily reversed bladder pain. Our findings suggest that spinal MIF and HMGB1 participate in persistent bladder pain induced by repeated intravesical PAR4 and may be potential therapeutic targets in chronic bladder pain conditions.

Protease activated-receptor 4 activation as a model of persistent bladder pain: Essential role of macrophage migration inhibitory factor and high mobility group box 1.

OBJECTIVES: To develop a rodent model of persistent non-inflammatory bladder pain and to test macrophage migration inhibitory factor and high mobility box group 1 as mediators of bladder pain. METHODS: Female C57BL/6 mice received intravesical instillations of protease activated receptor 4 (100 µmol/L, for 1 h) three times every other day and abdominal mechanical hypersensitivity (50% mechanical threshold) was tested on day 0 (baseline), and at days 1, 2, 3, 4, 7 and 9 after the first protease-activated receptor 4 injection. At the end of the experiment, micturition changes were measured and bladders were examined for histological changes. Macrophage migration inhibitory factor antagonist (MIF098; 40 mg/kg i.p. b.i.d.) or high mobility group box 1 inhibitor (glycyrrhizin; 50 mg/kg i.p. daily) was administered from day 2 until day 8. RESULTS: There was a significant and persistent decrease in abdominal mechanical threshold starting from day 3 in the protease-activated receptor 4-treated group that persisted until day 9 (5 days post-last instillation), but not in the control group. Glycyrrhizin fully reversed while MIF098 partially reversed abdominal mechanical hypersensitivity in protease-activated receptor 4-treated mice. The changes started on day 3 after the first protease-activated receptor 4 instillation, and analgesic effects lasted throughout the rest of the testing period. None of the groups had significant micturition changes or overt bladder histological changes. CONCLUSIONS: Repeated intravesical protease activated receptor 4 instillations produce persistent bladder pain without inflammation. Macrophage migration inhibitory factor and high mobility group box 1 are possible effective target molecules for bladder pain alleviation.

Elevated Urine Levels of Macrophage Migration Inhibitory Factor in Inflammatory Bladder Conditions: A Potential Biomarker for a Subgroup of Interstitial Cystitis/Bladder Pain Syndrome Patients.

OBJECTIVE: To investigate whether urinary levels of macrophage migration inhibitory factor (MIF) are elevated in interstitial cystitis/bladder pain syndrome (IC/BPS) patients with Hunner lesions and also whether urine MIF is elevated in other forms of inflammatory cystitis. METHODS: Urine samples were assayed for MIF by enzyme-linked immunosorbent assay. Urine samples from 3 female groups were examined: IC/BPS patients without (N = 55) and with Hunner lesions (N = 43), and non-IC/BPS patients (N = 100; control group; no history of IC/BPS; cancer or recent bacterial cystitis). Urine samples from 3 male groups were examined: patients with bacterial cystitis (N = 50), radiation cystitis (N = 18) and noncystitis patients (N = 119; control group; negative for bacterial cystitis). RESULTS: Urine MIF (mean MIF pg/mL ± standard error of the mean) was increased in female IC/BPS patients with Hunner lesions (2159 ± 435.3) compared with IC/BPS patients without Hunner lesions (460 ± 114.5) or non-IC/BPS patients (414 ± 47.6). Receiver operating curve analyses showed that urine MIF levels discriminated between the 2 IC groups (area under the curve = 72%; confidence interval 61%-82%). Male patients with bacterial and radiation cystitis had elevated urine MIF levels (2839 ± 757.1 and 4404 ± 1548.1, respectively) compared with noncystitis patients (681 ± 75.2). CONCLUSION: Urine MIF is elevated in IC/BPS patients with Hunner lesions and also in patients with other bladder inflammatory and painful conditions. MIF may also serve as a noninvasive biomarker to select IC/BPS patients more accurately for endoscopic evaluation and possible anti-inflammatory treatment.

Macrophage migration inhibitory factor mediates protease-activated receptor 4-induced bladder pain through urothelial high mobility group box 1.

Macrophage migration inhibitory factor (MIF) mediates pain although the mechanisms are not well understood. Urothelial activation of protease activated receptor 4 (PAR4) results in urothelial MIF release, urothelial high mobility group box 1 (HMGB1) release and bladder pain in mice without bladder inflammation. All three effects are prevented by MIF inhibition while intravesical disulfide HMGB1 alone can induce bladder pain. This study utilizes genetic MIF deletion to determine whether MIF mediates PAR4-induced bladder pain and is upstream of HMGB1-induced bladder pain. Wild type (C57/BL6) and MIF knockout (KO) mice were treated with intravesical PAR4 activating peptide or disulfide HMGB1 and tested for abdominal mechanical hypersensitivity at baseline (before treatment) and 24 h after injection. Micturition parameters and bladder histology were examined after behavioral test. Real-time PCR and western blotting measured HMGB1 mRNA and protein levels in the bladders of naïve wild type and MIF KO mice, while immunofluorescence measured HMGB1 protein levels in the urothelium of both strains. Intravesical PAR4 activation resulted in abdominal mechanical hypersensitivity in wild-type mice but not MIF KO mice. Intravesical disulfide HMGB1 induced abdominal mechanical hypersensitivity in both strains. Neither treatment resulted in significant changes in micturition or bladder histology in either strain. HMGB1 mRNA and protein levels were higher in MIF KO mouse bladders and the urothelium of MIF KO bladder had greater immunostaining than the wild-type strain. MIF is a pivotal molecule mediating PAR4-induced bladder pain and regulating urothelial HMGB1 production and release to elicit bladder pain.

Hereditary gingival fibromatosis: Characteristics and treatment approach.

Hereditary gingival fibromatosis (HGF) is a rare disorder characterized by a benign, non-hemorrhagic, fibrous gingival overgrowth that can appear in isolation or as part of a syndrome. Clinically, a pink gingiva with marked stippling can be seen to cover almost all the tooth, in many cases preventing eruption. HGF usually begins during the transition from primary to permanent teeth, giving rise to a condition that can have negative psychological effects at that age. As it does not resolve spontaneously, the treatment of choice is gingivectomy, which can be performed with an internal or external bevel incision, depending on each case and bearing in mind the changes that will take place at the dentogingival junction (DGJ). This paper describes clinical aspects and treatment in two eight-year-old boys with HGF, considering different facets of the surgical approach with conscious sedation in young children. Key words:Hereditary gingival fibromatosis, gingivectomy, internal bevel incision, external bevel incision, gingival overgrowth.

Disulfide high mobility group box-1 causes bladder pain through bladder Toll-like receptor 4.

BACKGROUND: Bladder pain is a prominent symptom in several urological conditions (e.g. infection, painful bladder syndrome/interstitial cystitis, cancer). Understanding the mechanism of bladder pain is important, particularly when the pain is not accompanied by bladder pathology. Stimulation of protease activated receptor 4 (PAR4) in the urothelium results in bladder pain through release of urothelial high mobility group box-1 (HMGB1). HGMB1 has two functionally active redox states (disulfide and all-thiol) and it is not known which form elicits bladder pain. Therefore, we investigated whether intravesical administration of specific HMGB1 redox forms caused abdominal mechanical hypersensitivity, micturition changes, and bladder inflammation in female C57BL/6 mice 24 hours post-administration. Moreover, we determined which of the specific HMGB1 receptors, Toll-like receptor 4 (TLR4) or receptor for advanced glycation end products (RAGE), mediate HMGB1-induced changes. RESULTS: Disulfide HMGB1 elicited abdominal mechanical hypersensitivity 24 hours after intravesical (5, 10, 20 µg/150 µl) instillation. In contrast, all-thiol HMGB1 did not produce abdominal mechanical hypersensitivity in any of the doses tested (1, 2, 5, 10, 20 µg/150 µl). Both HMGB1 redox forms caused micturition changes only at the highest dose tested (20 µg/150 µl) while eliciting mild bladder edema and reactive changes at all doses. We subsequently tested whether the effects of intravesical disulfide HMGB1 (10 µg/150 µl; a dose that did not produce inflammation) were prevented by systemic (i.p.) or local (intravesical) administration of either a TLR4 antagonist (TAK-242) or a RAGE antagonist (FPS-ZM1). Systemic administration of either TAK-242 (3 mg/kg) or FPS-ZM1 (10 mg/kg) prevented HMGB1 induced abdominal mechanical hypersensitivity while only intravesical TLR4 antagonist pretreatment (1.5 mg/ml; not RAGE) had this effect. CONCLUSIONS: The disulfide form of HMGB1 mediates bladder pain directly (not secondary to inflammation or injury) through activation of TLR4 receptors in the bladder. Thus, TLR4 receptors are a specific local target for bladder pain.

Protease-Activated Receptor 4 Induces Bladder Pain through High Mobility Group Box-1.

Pain is the significant presenting symptom in Interstitial Cystitis/Painful Bladder Syndrome (IC/PBS). Activation of urothelial protease activated receptor 4 (PAR4) causes pain through release of urothelial macrophage migration inhibitory factor (MIF). High Mobility Group Box-1 (HMGB1), a chromatin-binding protein, mediates bladder pain (but not inflammation) in an experimental model (cyclophosphamide) of cystitis. To determine if PAR4-induced bladder hypersensitivity depends on HMGB1 downstream, we tested whether: 1) bladder PAR4 stimulation affected urothelial HMGB1 release; 2) blocking MIF inhibited urothelial HMGB1 release; and 3) blocking HMGB1 prevented PAR4-induced bladder hypersensitivity. HMGB1 release was examined in immortalized human urothelial cultures (UROtsa) exposed to PAR4-activating peptide (PAR4-AP; 100 µM; 2 hours) or scrambled control peptide. Female C57BL/6 mice, pretreated with a HMGB1 inhibitor (glycyrrhizin: 50 mg/kg; i.p.) or vehicle, received intravesical PAR4-AP or a control peptide (100 µM; 1 hour) to determine 1) HMGB1 levels at 1 hour in the intravesical fluid (released HMGB1) and urothelium, and 2) abdominal hypersensitivity to von Frey filament stimulation 24 hours later. We also tested mice pretreated with a MIF blocker (ISO-1: 20 mg/kg; i.p.) to determine whether MIF mediated PAR4-induced urothelial HMGB1 release. PAR4-AP triggered HMGB1 release from human (in vitro) and mice (in vivo) urothelial cells. Intravesical PAR4 activation elicited abdominal hypersensitivity in mice that was prevented by blocking HMGB1. MIF inhibition prevented PAR4-mediated HMGB1 release from mouse urothelium. Urothelial MIF and HGMB1 represent novel targets for therapeutic intervention in bladder pain conditions.

Concentration measurement of yeast suspensions using high frequency ultrasound backscattering.

This work proposes the use of an ultrasound based technique to measure the concentration of yeasts in liquid suspension. This measurement was achieved by the detection and quantification of ultrasonic echoes backscattered by the cells. More specifically, the technique was applied to the detection and quantification of Saccharomyces cerevisiae. A theoretical approach was proposed to get the average density and sound speed of the yeasts, which were found to be 1116 kg/m(3) and 1679 m/s, respectively. These parameters were needed to model the waves backscattered by each single cell. A pulse-echo arrangement working around 50 MHz, being able to detect echoes from single yeasts was used to characterize experimentally yeast solutions from 10(2) to 10(7)cells/ml. The Non-negative Matrix Factorization denoising technique was applied for data analysis. This technique required a previous learning of the spectral patterns of the echoes reflected from yeasts in solution and the base noise from the liquid medium. Comparison between pulse correlation (without denoising) and theoretical and experimental pattern learning was made to select the best signal processing. A linear relation between ultrasound output and concentration was obtained with correlation coefficient R(2)=0.996 for the experimental learning. Concentrations from 10(4) to 10(7)cells/ml were detected above the base noise. These results show the viability of using the ultrasound backscattering technique to detect yeasts and measure their concentration in liquid cultures, improving the sensitivity obtained using spectrophotometric methods by one order of magnitude.