Leukocyte-endothelium interactions after hemorrhagic shock/reperfusion and cecal ligation/puncture: an intravital microscopic study in rat mesentery

Hemorrhagic shock/reperfusion (HS/R) followed by sepsis triggers systemic microcirculatory disturbances that may induce multiple organ failure. The present study evaluated the effects of HS/R and cecal ligation and puncture, followed by necrotic cecal resection/peritoneal lavage (REL) on leukocyte-endothelium interactions at the mesentery. Eighty-one anesthetized Wistar rats (200-250 g) were randomly assigned to a first injury: (1) control-HS-no hemorrhagic shock/no reperfusion group, (2) HS/blood-HS/R with 25% shed blood, and (3) HS/blood + LR-HS/R with 25% of the shed blood + lactated Ringer's solution, 3x shed blood volume. Twenty-four hours post-HS/R, animals were submitted to cecal ligation and puncture and, 24 h thereafter, to REL. Leukocyte-endothelium interactions were assessed by intravital microscopy and intercellular adhesion molecule (ICAM) 1 and P-selectin expression by immunohistochemistry. Lungs were observed for ICAM-1 expression and neutrophil infiltration. Single and double injury induced significant increases in rolling (approximately 2-fold), adherent (approximately 5-fold), and migrated leukocytes (approximately 7-fold); ICAM-1 expression (approximately 1/2-fold), and P-selectin expression (approximately 1/2-fold) at the mesentery compared with control-HS group. REL normalized leukocyte-endothelium interactions at the mesentery in single-injured animals. However, in double-injured rats, adherence and migration of leukocytes decreased but did not normalize. Similar results were observed on ICAM-1 expression and neutrophil infiltration in the lungs from these animals. In conclusion, the current in vivo observation of the mesenteric microcirculation after a double injury followed by REL is a suitable model for the systematic evaluation of the inflammatory reaction at local and distant sites. In addition, data presented herein emphasized the importance of surgical removal of the septic focus in controlling the otherwise lethal sepsis-induced multiple organ dysfunction syndrome.     

Increased risk of intrauterine transmission of HIV-1 associated with granulocyte elastase in endocervical mucus

BACKGROUND: One of the remaining challenges in the prevention of mother-to-child transmission of HIV is to reduce the risk of the transmission during pregnancy. It remains to be investigated which factors affect intrauterine HIV transmission and how they can be identified and addressed during pregnancy. METHODS: Granulocyte elastase in the endocervical mucus of HIV-positive pregnant women in Zambia was measured, and its association with intrauterine transmission of HIV-1 from the mother to the fetus was investigated. RESULTS: The intrauterine transmission rate determined by polymerase chain reaction assay of DNA from neonates at birth was 15.3%. The risk for intrauterine transmission was 8.65-fold higher in women who were positive for granulocyte elastase than in those who were negative. CONCLUSION: We suggest that the women showing positive granulocyte elastase at delivery be strongly suspected of having and if having had chorioamnionitis during pregnancy, which could affect the intrauterine transmission of HIV.     

Serum reactivity against bacterial pyruvate dehydrogenase: increasing the specificity of anti-mitochondrial antibodies for the diagnosis of primary biliary cirrhosis

Antimitochondrial antibodies (AMA) are the serum hallmark of primary biliary cirrhosis (PBC). However, AMA-positivity can be found in non-PBC sera when lower dilutions are used, thus raising issues about the specificity and sensitivity of the test. AMA reacts primarily with the lipoylated domains of pyruvate dehydrogenase-E2 (PDC-E2) which is highly conserved across species, including bacteria. We studied 77 serum samples, including 24 from patients with anti-PDC-E2-positive PBC and 53 controls (16 with autoimmune hepatitis (AIH), 10 with primary sclerosing cholangitis (PSC), and 27 healthy individuals) for their reactivities at serial dilutions (1:10, 1:20 and 1:40) against Escherichia coli DH5 alpha lysate overexpressing human PDC-E2 using immunoblotting (IB). A murine anti-human PDC-E2 monoclonal antibody (mAB) was used as control. We further studied positive sera using adsorption with a synthetic E. coli peptide sharing similarity with human PDC-E2. Finally, we verified whether a unique buffer for E. coli preparation could reduce non-specific serum reactivity. Results demonstrated that 100% of anti-PDC-E2-positive PBC and up to 38% of control sera at 1:10 dilution recognized E. coli PDC-E2 at IB while dilution tests indicated that the overall potency of PBC reactivity was 100-fold higher compared to controls. In fact, a subgroup (20-38%) of non-PBC sera were positive at low titers but lost the reactivity when absorbed with the synthetic E. coli peptide. Finally, our unique buffer reduced the reactivity of non-PBC sera as measured by ELISA. In conclusion, we demonstrated that weak cross-reactivity with E. coli PDC-E2 occurs in non-PBC sera at lower dilutions and that such reactivity is not due to AMA-positivity. The use of a specific buffer might avoid the risk of false positive AMA determinations when E. coli-expressed recombinant antigens are used.     

Chronic pain disorder manifesting as dental pain: case report

Chronic pain disorder is a challenging presentation for the dental surgeon, and carries considerable impairment for the affected individual. Patients undergo extensive unnecessary investigations and treatments before the correct diagnosis is reached. It is frequently a diagnosis of exclusion. We report the case of a 46-year-old man who experienced persistent severe orofacial pain superimposed upon pain involving other anatomical regions for more than 20 years. He received treatment by eight specialties and at five national centres with increasingly poor outcome.The importance of early recognition of medically unexplained pain will be discussed. Clinical Relevance: Dentists should have a high index of suspicion of pain disorder in those with multiple complaints of chronic pain in order to avoid unnecessary investigation and treatment.     

Assessment of sleep in women with night eating syndrome

STUDY OBJECTIVES: Evaluation of sleep in subjects with night eating syndrome (NES). DESIGN: Polysomnographic and questionnaire comparisons between subjects with NES and controls. SETTING AND PARTICIPANTS: Fifteen women with NES (mean +/- SD = 41 +/- 8 years) and 14 women (comparable age and weight) without NES (39 +/- 10 years) were studied in the laboratory for 3 days. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Subjects with NES did not differ from controls in timing of sleep onset or offset. They had less stage 2 sleep than controls (minutes, p = .012; percentage, p = .016) and less stage 3 sleep (p = .023), which contributed to their having a lower total sleep time (p = .05) and reduced sleep efficiency (p = .03). Subjects with NES did not have more awakenings than controls, but 93.3% of them ate on awakening during all 3 nights, while 92.9% of controls did not eat on any night. Logistic discriminant analyses identified a multiple sleep parameter model associated with increased likelihood of NES that had sensitivity of 84.6% and specificity of 76.9%. Patients with NES were more depressed than controls (p < .001) and reported greater sleep disturbance that included lower sleep quality (p < or = .001), reduced sleep duration (p < or = .001), and increased number of awakenings (p < or = .001). CONCLUSIONS: Patients with NES appear to have sleep maintenance insomnia rather than sleep-related eating disorder or a parasomnia. The maintenance of normal timing for sleep-wake behavior in the presence of a phase delay in the timing of caloric intake suggests this disorder reflects a state of internal circadian desynchrony associated with significant sleep complaints. It remains unknown whether the sleep disturbance precedes the abnormally timed eating.     

Equine estrogen-induced mammary tumors in rats

Long-term hormone replacement therapy is associated with an increased risk of breast, ovarian and endometrial cancers in women. Equine estrogens are a principal component of hormone replacement therapy; however, their tumorigenic potential toward mammary tissue and reproductive organs has not been extensively explored. A pellet containing equilin was inserted under the skin of female ACI rats and the development of mammary tumors was monitored. Histological examination revealed premalignant lesions such as apocrine metaplasia in whole-mount preparations of mammary gland from the equilin-treated rats. ACI rats given 10 mg equilin developed palpable mammary tumors at 13 weeks of treatment, and 37.5% of the rats developed mammary tumors within 15 weeks. For 2.5 mg equilin, palpable tumors were observed in 8.3% of the rats after 8 weeks’ treatment; the frequency was lower than that (42.9%) observed with 2.5 mg E₂. No tumors were observed in the untreated rats. Evidently, equilin is a mammary carcinogen, and this potential may be associated with development of breast and reproductive cancers in women receiving hormone replacement therapy.     

The OPCML tumor suppressor functions as a cell surface repressor-adaptor, negatively regulating receptor tyrosine kinases in epithelial ovarian cancer

Epithelial ovarian cancer is the leading cause of death from gynecologic malignancy, and its molecular basis is poorly understood. We previously demonstrated that opioid binding protein cell adhesion molecule (OPCML) was frequently epigenetically inactivated in epithelial ovarian cancers, with tumor suppressor function in vitro and in vivo. Here, we further show the clinical relevance of OPCML and demonstrate that OPCML functions by a novel mechanism in epithelial ovarian cancer cell lines and normal ovarian surface epithelial cells by regulating a specific repertoire of receptor tyrosine kinases: EPHA2, FGFR1, FGFR3, HER2, and HER4. OPCML negatively regulates receptor tyrosine kinases by binding their extracellular domains, altering trafficking via nonclathrin-dependent endocytosis, and promoting their degradation via a polyubiquitination-associated proteasomal mechanism leading to signaling and growth inhibition. Exogenous recombinant OPCML domain 1-3 protein inhibited the cell growth of epithelial ovarian cancers cell in vitro and in vivo in 2 murine ovarian cancer intraperitoneal models that used an identical mechanism. These findings demonstrate a novel mechanism of OPCML-mediated tumor suppression and provide a proof-of-concept for recombinant OPCML protein therapy in epithelial ovarian cancers. SIGNIFICANCE: The OPCML tumor suppressor negatively regulates a specific spectrum of receptor tyrosine kinases in ovarian cancer cells by binding to their extracellular domain and altering trafficking to a nonclathrin, caveolin-1–associated endosomal pathway that results in receptor tyrosine kinase polyubiquitination and proteasomal degradation. Recombinant OPCML domain 1-3 recapitulates this mechanism and may allow for the implementation of an extracellular tumor-suppressor replacement strategy.