Perioperative changes in cardiac biomarkers in juvenile cats during neutering.

Perioperative myocardial injury (PMI) is commonly caused by myocardial ischemia that develops during or after non-cardiac surgery. It occurs in 17.9% of human patients after non-cardiac surgery due to elevated high-sensitive perioperation cardiac troponin. However, PMI has not been demonstrated in cats. To investigate its occurrence, this study aimed to analyze the perioperative changes in cardiac biomarkers and clinical data, including measurement of vital signs, echocardiography, blood pressure, electrocardiogram, X-ray, and anesthetic profile, in 30 juvenile cats under neutering surgery. All cats had increased high-sensitive cardiac troponin I (hs-cTnI) postsurgery compared with presurgery. In particular, 48% of cats (14/29) showed elevated hs-cTnI over a reference range after surgery. In all groups, hs-cTnI and systolic arterial blood pressure (SAP) were significantly higher at 0 h and 18 h postoperation than at preoperation. A significant positive correlation was found between hs-cTnI and SAP at 18 h postoperation. Atrial natriuretic peptides, heart rate, and left ventricular wall thickness were markedly higher at 0 h postoperation than at preoperation; however, respiratory rate and body temperature were significantly lower at 0 h postoperation than at preoperation. Anesthetic time and hs-cTnI were significantly higher at 18 h postoperation in females than in males. Significant positive correlations were observed between hs-cTnI and anesthetic time at 18 h postoperation in females. These results indicate that postoperative hs-cTnI level can greatly increase in juvenile cats and hs-cTnI measurement at perioperation is potentially beneficial for early detection and evaluation of the presence of PMI.

Lipid Composition of Liposomal Membrane Largely Affects Its Transport and Uptake through Small Intestinal Epithelial Cell Models.

Lipid composition of liposomal bilayer should alter the cell response for permeability, transport, and uptake in small intestine. This work was done to investigate the transport and uptake of liposomes composed of docosahexaenoic acid-enriched phosphatidylcholine (PtdCho), phosphatidylserine (PtdSer), and sulfoquinovosyl diacylglycerol (SQDG) derived from marine products on multilamellar vesicles (MLV) in small intestinal epithelial cell models. The results showed that addition of PtdSer and SQDG as liposomal bilayer could improve the efficiency entrapment of liposomes. The liposomes containing PtdSer showed higher transport and uptake through both Caco-2 cell and M cell monolayers as compared to PtdCho-MLV. SQDG-containing liposomes exhibited only higher transport through M cell monolayer, while its uptake effect was higher both in Caco-2 cell and M cell monolayers. The results of experiments done with endocytosis inhibitors indicated that PtdCho-MLV must be transported via macropinocytosis and uptaken by phagocytosis in M cell monolayer model. PtdCho/PtdSer-MLV and PtdCho/SQDG-MLV might be transported and uptaken through M cell monolayer by phagocytosis. The result also indicated that PtdCho/SQDG-MLV could open the tight junction of small intestinal epithelial cell monolayers. Furthermore, our findings demonstrated that the surface status of cholesterol-containing liposomes were smooth, but they did not affect their transport and uptake through Caco-2 cell and M cell monolayers.

Distribution of regulatory T cells in inflammatory colorectal polyps of miniature dachshunds.

Inflammatory colorectal polyp (ICRP) is an emerging disease in Miniature Dachshunds (MDs). Animals with this disease exhibit multiple polyps with severe neutrophil infiltration that respond to immunosuppressive therapy. Macrophages in polypoid lesions have been described to play an important role in neutrophil infiltration in the lesion by producing IL-8. In contrast, IL-10, an anti-inflammatory cytokine, was also reported to be upregulated in polypoid lesions, but its significance in the pathogenesis of ICRP has not been clarified. Regulatory T cells (Tregs) are the main source of IL-10 production and contribute to the maintenance of intestinal homeostasis. Therefore, the objective of this research was to compare the distribution of Tregs in polypoid lesions of ICRPs and the association between the distribution and expression of pro- or anti-inflammatory cytokines. Tissue biopsy specimens of polypoid lesions were collected from 28 MDs with ICRP. Those of macroscopically non-polypoid colonic mucosa from 24 MDs with ICRPs and 21 control dogs were further included as controls. Real-time quantitative polymerase chain reaction was used to quantify gene expression of IL-1ß, IL-4, IL-6, IL-8, IL-10, IL-17, IL-22, IFN-γ, TNF-α, TGF-ß, and forkhead box protein P3 (Foxp3) in each tissue sample. The numbers of Foxp3-positive cells (Tregs) and ionized calcium binding adapter molecule 1 (Iba-1)-positive cells (macrophages) were determined by immunohistochemistry. The gene expression of IL-1ß, IL-6, IL-8, TNF-α, IFN-γ, IL-17, IL-10, TGF-ß, and Foxp3 was significantly upregulated in polypoid lesions relative to control levels. The numbers of Foxp3-positive Tregs and Iba-1-positive macrophages were significantly increased in polypoid lesions compared to those in the non-polypoid colonic mucosa of MDs with ICRPs and control dogs. The upregulation of IL-10 was moderately correlated with the distribution of Tregs in polypoid lesions from MDs with ICRPs. In addition, the relative upregulation of IL-1ß, IL-6, and IL-8 in polypoid lesions, compared to expression in non-polypoid colonic mucosa of MDs with ICRPs, was significantly greater than that of IL-10. These results indicate that increases in Treg numbers and anti-inflammatory cytokines in polypoid lesions comprise reactive changes in response to the inflammation, which warrants further investigation.

CD8⁺ tumor-infiltrating lymphocytes at primary sites as a possible prognostic factor of cutaneous angiosarcoma.

Tumor-infiltrating lymphocytes (TILs) have been reported as a prognostic factor in various cancers and are a promising target for immunotherapy. To investigate whether TILs have any impact on the prognosis of angiosarcoma patients, 55 non-treated patients (40 patients at stage 1 with cutaneous localized tumors, 4 patients at stage 2 with lymph node metastases and 11 patients at stage 3 with distant metastases) with angiosarcoma were evaluated retrospectively by immunohistochemistry stained CD4, CD8, FOXP3 and Ki67. The Kaplan-Meier method was used to estimate overall survival with patients at stage 1. Survival differences were analyzed by the log-rank test. Patients with higher numbers of CD8(+) TILs in their primary tumors survived significantly longer compared with patients with lower values. Moreover, the number of CD8 in TILs was positively correlated with a distant metastasis-free period. The total number of primary TILs (CD4 plus CD8) and CD8(+) primary TILs of stage 3 patients with distant metastases was positively correlated with their overall survival. To evaluate whether CD8(+) effector T cells are activated or differentiated, flow cytometric analysis of peripheral blood mononuclear cells (PBMC) was performed. The percentages of CD8(+) T cells producing IFN-γ in PBMC were significantly higher in patients with angiosarcoma (n = 10) compared not only with that of healthy controls (n = 20) but also patients with advanced melanoma (n = 11). These results suggest that anti-tumor immunity is clinically relevant in angiosarcoma.

Intractable diffuse alopecia caused by multifactorial side-effects in treatment of acute lymphocytic leukemia: connection to iatrogenic failure of estrogen secretion.

Treatment of infantile acute lymphocytic leukemia (ALL) may cause failure to thrive and hypogonadism due to hypopituitarism induced by chemotherapy and whole-brain radiotherapy. We report the case of a 22-year-old girl with a genetic predisposition to pattern hair loss who developed inveterate diffuse alopecia. The patient had onset of ALL at 8 years old and underwent bone marrow transplantation (BMT). Diffuse alopecia gradually advanced over her whole body. Her vellus scalp hair gradually came out, and hair loss progressed again at 8 years, after BMT. She later developed iatrogenic failure of secretion of estrogen and was treated with estrogen substitution therapy for 14 months from the age of 20. There was a small increase in the volume of hair during therapy, but alopecia returned to the former level after the therapy was suspended. Histopathologic examinations of the scalp performed during estrogen substitution therapy and 2 years after suspension of the therapy showed a 60% decrease in the number of hair follicles and prominent development of vellus hair. We conclude that estrogen influenced hair growth in the context of a genetic predisposition for pattern hair loss in this case.

Congenital vulvar lipoma within an accessory labioscrotal fold.

Perineal lipomas are uncommon congenital anomalies. They mainly develop in male patients with an accessory scrotum, posterior to the original scrotum. However, on rare occasions, these lipomas can occur in female patients with an accessory labioscrotal fold. We report on three such cases, where the lipomas were found in the anterior part of the right side of the labia majora. None had other anomalies, with the exception of one who had premature growth of pubic hair on the accessory labioscrotal fold. Skin with a rugated scrotal appearance was present in all cases.

[Anaphylactic reaction to latex during spinal anesthesia: a case report].

A 46-year-old man, with a history of atopic dermatitis and bronchial asthma, underwent surgery for an inguinal hernia. Forty-three minutes subsequent to spinal anesthesia, the patient complained suddenly of dyspnea with wheezing. Blood pressure decreased and skin eruption was observed on his chest. Postoperative laboratory tests revealed high IgE concentration, and a skin test confirmed an allergy to latex. The patient's allergic reaction was easily overlooked because of his history of bronchial asthma and the possibility that the hypotension was caused by the high spinal anesthesia. Latex allergy should be considered in any suspicious case presenting with these symptoms during surgery. After recovery, a skin test should be used to confirm the allergy to avoid repeated allergic episodes.

Erythemas caused by electrodes while monitoring neuromuscular blockade: three cases.

Skin erythemas formed in three patients during surgery at the sites where negative electrodes had been attached to stimulate the ulnar nerve for a neuromuscular transmission monitor (Relaxograph). The patients were all women, aged 52, 62, and 74 years, and general anesthesia lasted 8 h 20 min, 4 h 50 min, and 8 h 45 min, respectively. The electrodes used were disposable ECG electrodes in the first two patients and one designed for a neuromuscular monitor in the third; all were carbon-coated and then covered with gel. However, when the electrodes were detached from the lesion, they all showed loss or damage of the carbon coating under the gel. We recommend balancing the merit of monitoring with the risk of complications, even when applying an apparently safe, noninvasive monitor.

[A case of extensive Fournier's gangrene].

A 52-year-old man was admitted to our hospital complaining of high grade fever and painful swelling of the scrotum. Although debridement of the scrotum and inguinal region was immediately carried out, the inflammation extended to the right abdominal region 2 days later. We tunneled the newly infected areas and placed Penrose drains through the tunnels, so that complete drainage was achieved with minimal tissue loss. The infection was controlled by frequent irrigation of the wound and antibiotic administration. Forty days after the admission the defect of the skin was reconstructed by using a meshed skin graft, and the patient was discharged on the 68th hospital day. We concluded that limited debridement with placement of through-and-through drains might have produced a good result with minimal tissue loss.

New and better protocols for a short-term Caco-2 cell culture system.

The aim of the present study was to develop new and better protocols for a short-term Caco-2 cell culture system for use in rapid screening of intestinal drug absorption. Caco-2 cells were cultured according to several protocols for short-term cell culture to obtain monolayers. The effects of serum (fetal bovine serum, FBS) in the culture medium and of the period of cell culture on the barrier function and transporter activities of the monolayers were examined. The barrier function was estimated both from the transepithelial electrical resistance (TEER) and the permeability of [(14)C]mannitol. Transporter activities were monitored by measuring the permeability of [(14)C]glycylsarcosine for oligopeptide transporter (PepT1) and of rhodamine 123 for P-glycoprotein (P-gp). Caco-2 monolayers obtained by 3-day culture in the BIOCOAT HTS Caco-2 Assay System, developed by Becton Dickinson Bioscience, showed much higher permeability to hydrophilic compounds, such as mannitol, compared with those obtained by the standard 21-day culture system, due to the leaky structure of cell junctions. The newly developed 3-day protocol, which includes 10% FBS in the culture medium during the first day of culture, markedly enhanced TEER and lowered mannitol permeability of the monolayers. This protocol allowed us to better determine the rank order of permeability of compounds, giving results equivalent to those in the 21-day culture system. The longer culture period gave tighter monolayers, and the maximum value of TEER was obtained with 5 days in culture. However, after 5 days in culture, the integrity of monolayers decreased gradually. The highest activities of transporters, PepT1 and P-gp, in monolayers were obtained at days 5 or 6 of culture by the new protocol with FBS-containing medium. These results indicate that by a simple modification of the short-term culture protocol, it is possible to obtain Caco-2 monolayers with better barrier properties and higher activity of transporters that are equivalent to those found in the 21-day Caco-2 culture system.