High-resolution anoscopy, is there a benefit in proceeding directly to the operating room?

BACKGROUND: The development of high-resolution anoscopy (HRA) has advanced our ability to detect anal dysplasia. Historically, HRA is performed in a clinical setting and subsequent ablation is performed in the clinical setting or operating room. The aim of this study was to determine the most effective venue for the performance of HRA. METHODS: Following institutional review board (IRB) approval, the correlation between anal cytology and HRA performed in the clinic versus in the operating room was evaluated. Data were extracted from our IRB-approved prospective HRA database over the time period of 2013-2017. RESULTS: One hundred twenty-eight HRAs were compared (101 in the clinical setting, 27 in the operating room). There was a statistically significant difference in the correlation between anal cytology and HRA pathology for procedures performed in the clinical setting (55% [56/101]) versus those performed in the operating room (82% [22/27]) (p = 0.014). More biopsies were obtained in the operating room than in the clinic setting (3 vs. 1, p < 0.0001). The majority of patients who had HRA in a clinical setting with subsequent HRA in the operating room stated that they preferred to have their HRAs performed in the operating room due to discomfort from the HRA procedure. CONCLUSIONS: Detection rates for anal dysplasia on HRA, are significantly higher when performed in the operating room. To prevent discomfort in the clinical setting, patients with high-grade dysplasia on anal pap testing may benefit from proceeding directly to the operating room for concurrent HRA and ablation.

Monitoring in real time the effect of TLX overexpression on proliferation and migration of C6 cells.

Orphan nuclear receptor TLX has been shown to play an essential role in regulating the self-renewal and proliferation of neural stem cells (NSCs). However, TLX overexpression in NSCs induces long-term NSC expansion and further leads to glioma initiation in mouse when combined with p53 mutations. Whether overexpression of TLX plays a role in glioma stem cell (GSC) proliferation and migration still remains largely unknown. In this study, we infected C6 cells, a special glioma cell line which is mainly composed of cancer stem cells(CSCs), with lentiviruses expressing GFP(LV-GFP) or GFP-T2A-TLX(LV-TLX) and then monitored cell proliferation and migration using the real-time analyzer system (RTCA, xCELLigence, Roche). We found that the cell index (CI) observed for the TLX overexpressing C6 cells showed a lower value than that of the LV-GFP transduced cells. And the MTT results correlated highly with the RTCA proliferation assessments. Furthermore, the expression of p21 was decreased while other downstream genes PTEN and p53 were not significantly changed in TLX overexpressing C6 cells . These findings strongly indicate that TLX overexpression has the ability to decrease the proliferating and migratory properties of C6 cells by targeting p21. Further, our results suggest that TLX overexpression may also have a similar inhibitory effect on GSC proliferation and migration.

Risk factors for wound complications after abdominoperineal excision: analysis of the ACS NSQIP database.

AIM: The perineal wound following abdominoperineal excision (APR) is associated with a high complication rate. We aimed to evaluate the risk factors for wound complications and examine the effect of flap reconstruction on wound healing. METHOD: The American College of Surgeons National Surgical Quality Improvement Program (NSQIP) database was searched for patients who underwent APR for rectal adenocarcinoma. They were divided into two groups: primary closure of the perineal wound and flap reconstruction. A logistic regression analysis was performed to identify the risk factors for deep surgical site infection (SSI) and wound dehiscence. RESULTS: A total of 8449 (94%) patients from the database underwent primary closure and 550 (6%) underwent flap reconstruction. Patients who underwent flap reconstruction had a longer operation time, a higher incidence of deep SSI, wound dehiscence, more blood transfusion requirement and a higher rate of return to the operating room (all P < 0.001). Risk factors for deep SSI were African American race (OR 1.5, P = 0.02), American Society of Anesthesiologists (ASA) classification ≥ 4 (OR 3.2, P < 0.001), body mass index (BMI) ≥ 35 kg/m(2) (OR 1.7, P = 0.006), weight loss (OR 2, P < 0.001) and closure with a flap (OR 1.9, P < 0.001). Risk factors for wound dehiscence included ASA classification ≥ 4 (OR 2.2, P = 0.003), history of smoking (OR 2.2, P < 0.001), history of chronic obstructive pulmonary disease (OR 1.7, P = 0.03), BMI ≥ 35 kg/m(2) (OR 1.9, P = 0.001) and closure with a flap (OR 2.9, P < 0.001). CONCLUSION: Perineal wound complications are related to a patient's race, ASA classification, smoking, obesity and weight loss. Compared with primary closure, closure with a flap was associated with higher odds of wound infection and dehiscence and was not protective of wound complications in the presence of other risk factors. Therefore optimizing the patient's medical condition will lead to a better outcome irrespective of the technique used for perineal wound closure.

Effects of prolonged intensive training on the resting levels of salivary immunoglobulin A and cortisol in adolescent volleyball players.

AIM: Concerns have been raised regarding the effects of prolonged intensive training on adolescent athletes. This study investigated the differences in mucosal immune functions and stress responses between intensively trained male adolescent volleyball players and age-matched sedentary controls. METHODS: Twelve male volleyball players (16.5 [0.7] years of age) and sixteen healthy sedentary male volunteers (17.1 [0.6] years of age) participated in this study. Volleyball players were engaged in regular and year-round training. Unstimulated saliva samples were collected from volleyball players during the high-intensity training period and from the counterparts at the same timepoints after at least 18 hours of rest. Concentrations of salivary total protein, secretory immunoglobulin A (SIgA), cortisol, and lactoferrin were measured. RESULTS: Results of this study revealed that the SIgA concentrations and the ratio of SIgA/total protein in volleyball players were significantly lower compared with those in sedentary controls. However, the salivary cortisol concentrations and the ratio of cortisol/total protein in volleyball players were markedly higher compared with those in sedentary controls. No significant difference was observed in lactoferrin levels between volleyball players and sedentary controls. CONCLUSION: The findings of this study suggest that the prolonged intensive training may elicit a sustained stress and induce a suppressive effect on mucosal immunity in regularly and intensively trained adolescent athletes.

Impact of intense training and rapid weight changes on salivary parameters in elite female Taekwondo athletes.

The aim of this study is to examine the cumulative effects of prolonged intensive training with or without rapid weight changes (RWC) on salivary parameters of elite female Taekwondo (TKD) athletes. Ten elite female Taiwanese TKD athletes (ages: 21.3 ± 1.2 years of age, Ht 164.4 ± 5.6 cm) volunteered to participate in this study. Resting saliva samples were collected at 28-, 14-, 7-, and 1 day before and 1-, 7-, 21 days after a national competition. The levels of salivary immunoglobulin A (sIgA), cortisol, and lactoferrin were measured. In analyzing the anthropometric data, we found that a significant proportion (50%) of elite female TKD athletes had RWC shortly before and after a national competition. The participants were allocated either to the RWC or to the non-RWC group according to their weight change profiles. Our results showed that levels of sIgA and cortisol of athletes with RWC were significantly modulated during the study period. However, athletes without RWC only showed reduced lactoferrin after competition. The results presented here demonstrate that intensive training in combination with RWC affects the mucosal immunity and disrupts the cortisol stress response of elite female TKD athletes.

Changes of mucosal immunity and antioxidation activity in elite male Taiwanese taekwondo athletes associated with intensive training and rapid weight loss.

OBJECTIVE: The aim of this study was to investigate the cumulative effects of prolonged, intensive training and rapid weight loss on immunological parameters and antioxidation activity of elite male Taiwanese taekwondo athletes. DESIGN: 16 Elite male taekwondo athletes (mean age, 21.6 (1.3) years; mean height, 173.7 (5.5) cm) volunteered to participate in this study. Beginning at 30 days before a national competition, saliva samples were obtained during a 7-week training, the competition and the postcompetition period. Levels of salivary IgA, cortisol, lactoferrin and free-radical scavenging activity were measured at 30-, 14-, 7- and 1-day precompetition and 1-, 7- and 19-day postcompetition. Body weight and body fat were also recorded. RESULTS: The mean body weight was notably decreased during the week immediately before the competition. Results reveal that the levels of salivary IgA were differentially regulated during the training, competition and recovery period, while the salivary cortisol and lactoferrin concentrations and free-radical scavenging activity were not appreciably affected during the training and the competition period. Furthermore, the results of an upper respiratory tract infection incidence indicate that following the decreases of mucosal immunity, the risk of acquiring infection was significantly increased. CONCLUSIONS: Our results demonstrated that mucosal immunity in elite male taekwondo athletes is modulated by exercise and rapid weight reduction during the training, competition and recovery period. Cumulative effects of prolonged intensive training and rapid weight reduction suppressed mucosal immunity. Furthermore, because of the "open window" of impaired immunity during the precompetition period, the incidence of upper respiratory tract infection was significantly increased after the competition.

MiR-15b ameliorates SONFH by targeting Smad7 and inhibiting osteogenic differentiation of BMSCs.

OBJECTIVE: The aim of this study was to explore the role of micro ribonucleic acid (miR)-15b in steroid-induced osteonecrosis of the femoral head (SONFH) and its potential mechanism. PATIENTS AND METHODS: Bone marrow tissues were collected from 5 patients with glucocorticoid (GC)-induced ONFH (GC-ONFH, GC group) and 5 patients with secondary ONFH (control group) undergoing total hip replacement in our hospital from July 2016 to August 2017. Subsequently, bone marrow mesenchymal stem cells (BMSCs) were separated from bone marrow extracted and cultured in vitro. Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) assay was used to detect differentially expressed miRNAs in BMSCs of patients in GC group and control group. BMSCs were treated with different concentrations of GC. Next, the effect of GC on tmiR-15b expression level was detected via qRT-PCR. Alizarin red staining assay was performed to evaluate the effect of miR-15b on osteogenic differentiation of BMSCs. Meanwhile, the potential targets of miR-15b were predicted using bioinformatics software and validated through luciferase reporter gene assay, respectively. Additionally, Western blotting was conducted to determine the effect of miR-15b on the protein expression of the transforming growth factor beta (TGF-ß) signaling pathway. RESULTS: Flow cytometry demonstrated that the proportion of cluster of differentiation 44 (CD44)-positive cells was 99.7%, while that of CD45-positive cells was only 0.17% in cultured BMSCs. This suggested that the purity of BMSCs was relatively high. QRT-PCR assay indicated that the expression level of miR-15b declined significantly in BMSCs of GC group when compared with control group (p<0.01). The osteogenic differentiation capacity of BMSCs was significantly strengthened in GC group compared with control group (p<0.01). Subsequent qRT-PCR assay revealed that GC down-regulated the expression level of miR-15b in a dose-dependent manner. Besides, the osteogenic differentiation capacity of cells was remarkably strengthened in miR-15b mimic treatment group when compared with control group (p<0.01). Bioinformatics software (TargetScan) predicted that drosophila mothers against decapentaplegic protein 7 (Smad7) might be a potential target of miR-15b, which was indicated by luciferase reporter gene assay. In comparison with control group, miR-15b mimic treatment group exhibited significantly down-regulated protein expression level of Smad7, increased expression level of phosphorylated (p)-Smad2/3 and up-regulated messenger RNA (mRNA) expression level of runt-related transcription factor 2 (Runx2). However, the protein expression level of Smad7 and p-Smad2/3 and the mRNA expression level of Runx2 exhibited opposite trends in miR-15b inhibitor treatment group. CONCLUSIONS: MiR-15b relieves SONFH by targeting Smad7 and repressing osteogenic differentiation of BMSCs.

Increased expression of cysteinyl leukotriene receptor-1 in the brain mediates neuronal damage and astrogliosis after focal cerebral ischemia in rats.

Cysteinyl leukotrienes are potent pro-inflammatory mediators. Cysteinyl leukotriene receptor 1 is one of the two cysteinyl leukotriene receptors cloned. We recently reported that cysteinyl leukotriene receptor 1 antagonists protected against cerebral ischemic injury, and an inducible expression of cysteinyl leukotriene receptor 1 was found in neuron- and glial-appearing cells after traumatic injury in human brain. To determine the role of cysteinyl leukotriene receptor 1 in ischemic brain injury, we investigated the temporal and spatial profile of cysteinyl leukotriene receptor 1 expression in rat brain from 3 h to 14 days after 30 min of middle cerebral artery occlusion, and observed the effect of pranlukast, a cysteinyl leukotriene receptor 1 antagonist, on the ischemic injury. We found that cysteinyl leukotriene receptor 1 mRNA expression was up-regulated in the ischemic core both 3-12 h and 7-14 days, and in the boundary zone 7-14 days after reperfusion. In the ischemic core, cysteinyl leukotriene receptor 1 was primarily localized in neurons 24 h, and in macrophage/microglia 14 days after reperfusion; while in the boundary zone it was localized in proliferated astrocytes 14 days after reperfusion. Pranlukast attenuated neurological deficits, reduced infarct volume and ameliorated neuron loss in the ischemic core 24 h after reperfusion; it reduced infarct volume, ameliorated neuron loss and inhibited astrocyte proliferation in the boundary zone 14 days after reperfusion. Thus, we conclude that cysteinyl leukotriene receptor 1 mediates acute neuronal damage and subacute/chronic astrogliosis after focal cerebral ischemia.

HAMI 3379, a CysLT2R antagonist, dose- and time-dependently attenuates brain injury and inhibits microglial inflammation after focal cerebral ischemia in rats.

Cysteinyl leukotrienes (CysLTs) induce inflammatory responses by activating their receptors, CysLT1R and CysLT2R. We have reported that CysLT2R is involved in neuronal injury, astrocytosis, and microgliosis, and that intracerebroventricular (i.c.v.) injection of the selective CysLT2R antagonist HAMI 3379 protects against acute brain injury after focal cerebral ischemia in rats. In the present study, we clarified features of the protective effect of intraperitoneally-injected HAMI 3379 in rats. We found that HAMI 3379 attenuated the acute brain injury 24 h after middle cerebral artery occlusion (MCAO) with effective doses of 0.1-0.4 mg/kg and a therapeutic window of ∼1h. It attenuated the neurological deficits, and reduced infarct volume, brain edema, and neuronal loss and degeneration 24 and 72h after MCAO. RNA interference with i.c.v. injection of CysLT2R short hairpin RNA (shRNA) attenuated the acute injury as well. Also, HAMI 3379 inhibited release of the cytokines IL-1ß, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) into the serum and cerebrospinal fluid 24h after MCAO. Moreover, HAMI 3379 ameliorated the microglial activation and neutrophil accumulation in the ischemic regions, but did not affect astrocyte proliferation 72h after MCAO. In comparison, the CysLT1R antagonist pranlukast did not affect microglial activation and IFN-γ release, but inhibited astrocyte proliferation and reduced serum IL-4. Thus, we conclude that HAMI 3379 has a protective effect on acute and subacute ischemic brain injury, and attenuates microglia-related inflammation. CysLT2R antagonist(s) alone or in combination with CysLT1R antagonists may be a novel class of therapeutic agents in the treatment of ischemic stroke.

Evidence for arylamine N-acetyltransferase activity in the fungi Candida albicans.

N-acetyltransferase activities were determined in Candida albicans, which is a member of the normal flora of the mucous membranes in the respiratory, gastrointestinal and female genital tract. The N-acetylation of 2-aminofluorene and p-aminobenzoic acid by the N-acetyltransferase from Candida albicans was determined using high pressure liquid chromatography. The activities (mean +/- S.D.) of N-acetyltransferase from Candida albicans cytosols were 1.06 +/- 0.01 nmol/min per mg protein for the acetylation of 2-aminofluorene substrate, and not detectable levels of acetyl-p-aminobenzoic acid for the acetylation of p-aminobenzoic acid. The apparent kinetic constants Km and Vmax values were 0.17 +/- 0.06 mM and 1.43 +/- 0.42 nmol/min per mg protein, respectively, for 2-aminofluorene substrate. The optimum pH value for the enzyme activity was 8.0. The optimal temperature for the enzyme activity is 40 degrees C for 2-aminofluorene substrate. Among a series of divalent cations and salts, Fe2+, SCN-, I-, and NH4+ were demonstrated to be the most potent inhibitors. The N-acetyltransferase activity was inhibited by iodoacetamide: at 0.25 mM iodoacetamide, activity was reduced 50% and 1.0 mM iodoacetamide inhibited activity more than 90%. This is the first demonstration of acetyl CoA arylamine N-acetyltransferase activity in the yeast-like fungus Candida albicans.

Relationship between development of intramuscular connective tissue and toughness of pork during growth of pigs.

We investigated changes in structures and properties of the endomysium and perimysium during development of semitendinosus muscle in relation to the increase in toughness of pork using samples from neonates to 55-mo-old pigs. The shear force value of pork increased linearly until 6 mo of age, and the rate of increase slowed down thereafter. The secondary perimysium thickened owing to an increase in the number and thickness of perimysial sheets consisting of collagen fibers, which became thicker and wavy with the growth of the pigs. This increase in thickness of the secondary perimysium was correlated significantly with the increase in the shear force value (r = .98). The endomysial sheaths became thicker and denser in the muscle of 6-mo-old pigs. Maturation of the endomysium was accompanied by hypertrophy of muscle fibers. The amount of heat-soluble collagen decreased almost linearly, indicating that nonreducible cross-links between collagen molecules were formed throughout chronological aging. We conclude that thickening of the perimysium is closely related to an increase in the toughness of pork during growth of pigs.

Mechanical test method on the estimation of the lubricant performance by hyaluronic acid.

Cases with loss of articular cartilage or lowering of articular lubricant performance are frequent among patients afflicted by various gonarthropathies or those fitted with prosthetic joints. The management of such cases has necessitated evaluation studies on the clinical efficacy of using high molecular weight hyaluronic acid as an artificial synovial fluid with a view to improving articular lubricant performance. The proper implementation of such evaluation studies on lubrication performance requires testing methods capable of assessing basic lubricant performance not only with respect to articular cartilage alone but also with respect to various other combinations of materials, i.e., lubrication between articular bone and artificial joints, etc. In the present study, first, a simple standard lubricant performance test using several small readily procurable test specimens was devised. Then, this test was employed to evaluate the basic lubricant properties of hyaluronic acid intended for use as artificial synovial fluid.

The new P2Y-like receptor G protein-coupled receptor 17 mediates acute neuronal injury and late microgliosis after focal cerebral ischemia in rats.

G protein-coupled receptor 17 (GPR17), the new P2Y-like receptor, is phylogenetically related to the P2Y and cysteinyl leukotriene receptors, and responds to both uracil nucleotides and cysteinyl leukotrienes. GPR17 has been proposed to be a damage sensor in ischemic stroke; however, its role in brain inflammation needs further detailed investigation. Here, we extended previous studies on the spatiotemporal profiles of GPR17 expression and localization, and their implications for brain injury after focal cerebral ischemia. We found that in the ischemic core, GPR17 mRNA and protein levels were upregulated at both 12-24 h and 7-14 days, but in the boundary zone the levels increased 7-14 days after reperfusion. The spatiotemporal pattern of GPR17 expression well matched the acute and late (subacute/chronic) responses in the ischemic brain. According to previous findings, in the acute phase, after ischemia (24 h), upregulated GPR17 was localized in injured neurons in the ischemic core and in a few microglia in the ischemic core and boundary zone. In the late phase (14 days), it was localized in microglia, especially in activated (ED1-positive) microglia in the ischemic core, but weakly in most microglia in the boundary zone. No GPR17 was detectable in astrocytes. GPR17 knockdown by a small interfering RNA attenuated the neurological dysfunction, infarction, and neuron loss at 24 h, and brain atrophy, neuron loss, and microglial activation at 14 days after reperfusion. Thus, GPR17 might mediate acute neuronal injury and late microgliosis after focal cerebral ischemia.

Cysteinyl leukotriene receptor 2 is spatiotemporally involved in neuron injury, astrocytosis and microgliosis after focal cerebral ischemia in rats.

Cysteinyl leukotrienes (CysLTs), potent inflammatory mediators, are released from ischemic brain, and may regulate ischemic injury through activating CysLT1 and CysLT2 receptors. The CysLT1 receptor is closely associated with ischemic injury and post-ischemic repair; however, the CysLT2 receptor-mediated responses remain unknown. Here, we investigated the spatiotemporal profiles and implications of CysLT2 receptor expression and localization in rat brain after focal cerebral ischemia. CysLT2 receptors were normally localized in astrocytes in the cortex and around the ventricles. After focal cerebral ischemia, CysLT2 receptor expression was up-regulated in concert with neuronal and glial responses. In the acute phase (6-24 h), up-regulated CysLT2 receptors were restricted to injured neurons in the ischemic core; while in the late phase (3-28 days), the up-regulation was restricted to hypertrophic microglia (ischemic core) and mainly localized in hypertrophic astrocytes (boundary zone). Thus, the spatiotemporal profiles of CysLT2 receptor expression suggest that it plays regulatory roles in acute neuron injury, and astrocytosis and microgliosis in the late phase.

Morin sulphates/glucuronides enhance macrophage function in microgravity culture system.

BACKGROUND: The immune system changes significantly in astronauts during and after space flight. Although the mechanism has not been defined, it is reasonable to begin developing effective countermeasures to the physiological consequences of spaceflight, especially immunosuppression. Many studies have been published about the effect of flavonoids on immune modulation. Thus, the aim of this study was to develop whether flavonoids could be the effective countermeasures to the immunosuppression caused by microgravity. MATERIALS AND METHODS: We used a rotating wall vessel 3D (three-dimensional) culture system which recreates some of the culture conditions that occur during microgravity to study the effects of microgravity on the function of macrophages and assess the modulating effects of flavonoids on microgravity-induced macrophage dysfunction. RESULTS: We demonstrated 65% and 80% reduction in mitogen-induced nitric oxide and cytokine production of 3D-cultured macrophages, compared to conventional two-dimensional (2D)-cultured cells. Moreover, the microgravity-induced macrophage dysfunction was not restored by transferring cells from 3D to 2D culture. However, the addition of morin sulphates/glucuronides in 3D culture compensated for the loss of macrophage function. CONCLUSION: The result presented here suggests for the first time that an immune-modulatory strategy using flavonoid supplements such as morin would benefit the health of astronauts.

The self-cleaving activity of nucleolin determines its molecular dynamics in relation to cell proliferation.

Nucleolin (105 kDa) is the most abundant nucleolar phosphoprotein in exponentially growing cells. We have demonstrated that nucleolin has an intrinsic protease activity for autodegradation. This self-cleaving activity is markedly decreased when cells enter the proliferative stage, resulting in significant stabilization of the nucleolin molecule. The fusion protein derived from a partial nucleolin cDNA clone, which encodes the C-terminal two-thirds of nucleolin, maintained the self-cleaving activity similar to that of the nucleolin purified from nonproliferating cells. The enzymatic domain is therefore located within this region. An exogenous peptide, highly negatively charged, exerted an inhibitory effect, indicating that the enzymatic activity can be regulated. The cleavage kinetics of nucleolin isolated from proliferating cells was apparently concentration independent, suggesting that the autoproteolytic reaction of nucleolin in cells at dividing stage is an intramolecular event. There were many cleavage fragments, which were clustered at four major sites, with apparent molecular weights of about 100, 70, 60, and 50 kDa. These results imply that the nucleolin molecule has repeated motifs that serve as the cleavage sites. Since the detailed electrophoresis patterns, including location and intensity, of the in vitro self-cleaved products of nucleolin derived from nonproliferating and proliferating cells were not quite the same, preferential usage of the particular cutting sites according to cell stage was suggested. It is possible that post-translational modification of nucleolin in proliferating cells may cause these variations.