Presence of Human Papillomavirus and Epstein-Barr Virus in Squamous Lesions of the Tongue

ABSTRACT Background: Several studies have suggested a possible role of human papillomavirus (HPV) and Epstein-Barr virus in the pathogenesis of oral premalignant lesions. This study aimed to investigate the correlation between squamous dysplasia of the tongue and expression of p16 and Ki67 immunohistochemically as well as HPV genotypes with real-time polymerase chain reaction (PCR). Methods: Twenty-three tongue biopsies were stained immunohistochemically for p16, Epstein- Barr virus and Ki67 and real-time PCR and chromogenic in-situ hybridization for HPV. Results: Dysplasia was diagnosed in 16 of 23 cases without invasive carcinoma and suspicious for dysplasia (n=17) and HPV infection (n=6). These were subjected to chromogenic in-situ hybridization for HPV DNA (HPV-III family 16). There was no immunoreactivity for Epstein-Barr virus. p16 was positive in 4/16 (25%) of dysplastic lesions. One lesion was positive for HPV by chromogenic in-situ hybridization, and one case was positive by real-time PCR for HPV. Conclusion: This evidence suggested that HPV infection but not Epstein-Barr virus infection plays a role in pathogenesis of squamous dysplasia localized tongue.

The anti-inflammatory and antioxidant effects of thymoquinone on ceruleine induced acute pancreatitis in rats.

INTRODUCTION: The aim of this study is to investigate the potential antioxidant and anti-inflammatory effects of thymoquinone (TQ) on ceruleine induced acute pancreatitis. MATERIAL AND METHODS: A total of 14 male Wistar albino rats were divided into 2 groups as follows: (1) normal saline-treated group and (2) thymoquinone- treated groups. For achieving acute pancreatitis, intraperitoneal (IP) cerulein, a stable cholecystokinin (CCK) analogue, was applied in a 50 mcg/kg dose 2 times in one-hour interval in total. One hour after last ceruleine injection, IP 2 ml/kg isotonic saline solution was applied to the saline group and IP 5 mg/kg TQ was applied. The rats were sacrificed by decapitation 12 h after the last injection of last medication. Blood samples were taken, and serum interleukin-1ß (IL-1ß), amylase, lipase pancreatic, total antioxidant capacity (TAC), total oxidant status (TOS), and pancreatic Schoenberg scores were determined. Oxidative stress index (OSI) was calculated for each group. Results are given as mean ± SD. A value of p < 0.05 was accepted as statistically significant. SPSS for Windows v15.0 was used for statistical analyses. RESULTS: The increased serum amylase, lipase levels and histopathological scoring of pancreatic tissue showed that acute pancreatitis was present in both groups. Furthermore, serum IL-1ß level was significantly reduced in TQ administered group (p < 0.05). Although serum TAC level was high and TOS level was low, those changes were not statistically significant. Nevertheless, OSI index, which was driven from TOS/TAC, was significantly low in TQ groups (p < 0.05). Although TQ partially ameliorated the acute pancreatitis in terms of histopathological evaluations, the main effect of it was brought about by reducing the hemorrhage in acute pancreatitis (p < 0.05). CONCLUSION: In this study, it was shown that TQ can reduce the inflammation and has a positive effect on the oxidative status of organism in inflammatory cases such as acute pancreatitis. This is consistent with partial amelioration of acute pancreatitis in rats given TQ (Tab. 2, Fig. 4, Ref. 31).

Association of the expression levels in the longissimus muscle and a SNP in the CDC10 gene with marbling in Japanese Black beef cattle.

The septin 7 (CDC10) gene, involved in cellular proliferation, has been previously shown to be expressed at different levels in the longissimus muscle (LM) between low-marbled and high-marbled steer groups by differential-display PCR. It is located within the genomic region of a quantitative trait locus for marbling, and thus was considered as a positional functional candidate gene for marbling. In this study, we showed that the CDC10 expression levels in the LM were positively correlated with marbling in Japanese Black (JB) steers (P<0.0001). Further, an association analysis indicated that a SNP in the promoter region of the CDC10 gene was associated with marbling using 99 JB sires (P=0.03) and 542 JB paternal half-sib progeny steers from a sire homozygous for G allele at the SNP (P<0.0001). These findings suggest possible effects of the expression levels in the LM and the SNP of the CDC10 gene on marbling in JB cattle.

Association of the expression levels in the skeletal muscle and a SNP in the CDC10 gene with growth-related traits in Japanese Black beef cattle.

Growth performance, as well as marbling, is the main breeding objective in Japanese Black (JB) cattle, the major beef breed in Japan. The septin 7 (CDC10) gene, involved in cellular proliferation, is located within a genomic region of a quantitative trait locus for growth-related traits. In this study, we first showed that the expression levels of the CDC10 gene in the skeletal muscle were higher in JB steers with extremely high growth performance than in JB steers with extremely low growth, using real-time PCR. Further, a single nucleotide polymorphism (SNP), NC_007302.5:g.63264949G>C, was detected in the promoter region of the CDC10 gene and genotyped in three Japanese cattle breeds (known as 'Wagyu' in Japan) and the Brown Swiss dairy cattle breed. All four cattle populations showed a moderate genetic diversity at the SNP of the CDC10 gene. An association analysis indicated that the SNP was associated with growth-related traits in JB cattle. These findings suggest possible effects of the expression levels in the skeletal muscle and the SNP of the CDC10 gene on growth-related traits in JB cattle. The CDC10 SNP may be useful for effective marker-assisted selection to increase beef productivity in JB beef cattle.

Cuprizone-induced demyelination in Wistar rats; electrophysiological and histological assessment.

OBJECTIVES: Multiple Sclerosis (MS) is a disease that affects the Central Nervous System by destructing myelin shield and also can affects the peripheral nervous system. Demyelination is acquired characteristics disease and appears with the degeneration of myelin which protects the axons. Cuprizone (CPZ) model is a toxic demyelination model. The purpose of this study was to develop an MS model by cuprizone exposure to Wistar rats. MATERIALS AND METHODS: Rats were separated into control and experimental groups and daily cuprizone was administered to experimental groups for 4, 5, 6 and 7 weeks. At the end of the experiments, spinal nerve conduction velocity was measured by EMG detected from the gastrocnemius muscle. After scarification, cerebrum and cerebellum of the animals were taken for histopathological investigation. RESULTS: Spinal cord nerve conduction velocity (SNCV) of control animals was 76.54 m/s. Whereas SNCV of the rats that were feed with CPZ for 6 weeks was significantly reduced to 46.35 m/s in comparison with the control group. Demyelinated areas and vacuolization were seen on the brain sections of CPZ exposed rats. CONCLUSIONS: SNCV of the rats were feed with cuprizone began tendency of decrease after 4th weeks. These reductions were observed as maximum at 6th weeks. At 7th week increments were observed at SNCV. These results indicated that 6 weeks of cuprizone feedings could be suitable to bring into existence of MS model in Wistar rats.

Risk factors for early bacterial infections in liver transplantation.

OBJECTIVE: Our aim was to determine perioperative risk factors for early bacterial infection after liver transplantation. METHODS: Retrospectively examining medical records using Centers for Disease Control and Prevention (CDC) definitions to identify nosocomial infections, we analyzed data on 367 adult patients. RESULTS: The incidence of infection was 37.3% (n = 137): namely, surgical site (n = 74; 20.2%) [corrected], blood stream (n = 64; 17.4%), pulmonary (n = 49; 13.4%), urinary system (n = 26; 7.1%). Significant risk factors within the first 30 days were as follows: deceased donor, Model for End-Stage Liver Disease (MELD) >20, albumin level <2.8 g/dL, intraoperative erythrocyte transfusion >6 U, intraoperative fresh frozen plasma >12 U, bilioenteric anastomosis, postoperative intensive care unit stay >6 days, and postoperative length of stay >21 days. Significant risk factors detected within the first 90 days were as follows: MELD >20, preoperative length of stay >7 days, reoperation, postoperative length of intensive care unit stay >6 days, and postoperative length of stay >21 days. Variability was observed in risk factors according to localization of infection. As a result, except for MELD, type of donor, and biliary anastomosis, the others are preventable factors for early bacterial infection. In addition, the same risk factors showed variability according to the site of infection.

Adsorption kinetics of aromatic compounds on carbon nanotubes and activated carbons.

Adsorption kinetics of two organic compounds on four types of carbonaceous adsorbents (a granular activated carbon [HD4000], an activated carbon fiber [ACF10], a single-walled carbon nanotube [SWNT], and a multiwalled carbon nanotube [MWNT]) was examined in aqueous solutions. The times needed for the adsorption to reach apparent equilibrium on the four carbons followed the order of ACF10 > HD4000 > SWNT > MWNT. Ultrasonication of the carbon nanotubes (CNTs) accelerated their adsorption kinetics but had no effect on their equilibrium adsorption capacities. The pseudo-second order model (PSOM) provided good fitting for the kinetic data. The fitting of kinetic data with the intraparticle diffusion model indicated that external mass transfer controls the sorption process in the organic compound-CNT systems, whereas intraparticle diffusion dominates in the sorption of organic compounds onto activated carbons.

Learning-stage-dependent, field-specific, map plasticity in the rat auditory cortex during appetitive operant conditioning.

Cortical reorganizations during acquisition of motor skills and experience-dependent recovery after deafferentation consist of several distinct phases, in which expansion of receptive fields is followed by the shrinkage and use-dependent refinement. In perceptual learning, however, such non-monotonic, stage-dependent plasticity remains elusive in the sensory cortex. In the present study, microelectrode mapping characterized plasticity in the rat auditory cortex, including primary, anterior, and ventral/suprarhinal auditory fields (A1, AAF, and VAF/SRAF), at the early and late stages of appetitive operant conditioning. We first demonstrate that most plasticity at the early stage was tentative, and that long-lasting plasticity after extended training was able to be categorized into either early- or late-stage-dominant plasticity. Second, training-induced plasticity occurred both locally and globally with a specific temporal order. Conditioned-stimulus (CS) frequency used in the task tended to be locally over-represented in AAF at the early stage and in VAF/SRAF at the late stage. The behavioral relevance of neural responses suggests that the local plasticity also occurred in A1 at the early stage. In parallel, the tone-responsive area globally shrank at the late stage independently of CS frequency, and this shrinkage was also correlated with the behavioral improvements. Thus, the stage-dependent plasticity may commonly underlie cortical reorganization in the perceptual learning, yet the interactions of local and global plasticity have led to more complicated reorganization than previously thought. Field-specific plasticity has important implications for how each field subserves in the learning; for example, consistent with recent notions, A1 should construct filters to better identify auditory objects at the early stage, while VAF/SRAF contribute to hierarchical computation and storage at the late stage.

Adsorption of aromatic compounds by carbonaceous adsorbents: a comparative study on granular activated carbon, activated carbon fiber, and carbon nanotubes.

Adsorption of three aromatic organic compounds (AOCs) by four types of carbonaceous adsorbents [a granular activated carbon (HD4000), an activated carbon fiber (ACF10), two single-walled carbon nanotubes (SWNT, SWNT-HT), and a multiwalled carbon nanotube (MWNT)] with different structural characteristics but similar surface polarities was examined in aqueous solutions. Isotherm results demonstrated the importance of molecular sieving and micropore effects in the adsorption of AOCs by carbonaceous porous adsorbents. In the absence of the molecular sieving effect, a linear relationship was found between the adsorption capacities of AOCs and the surface areas of adsorbents, independent of the type of adsorbent. On the other hand, the pore volume occupancies of the adsorbents followed the order of ACF10 > HD4000 > SWNT > MWNT, indicating that the availability of adsorption site was related to the pore size distributions of the adsorbents. ACF10 and HD4000 with higher microporous volumes exhibited higher adsorption affinities to low molecular weight AOCs than SWNT and MWNT with higher mesopore and macropore volumes. Due to their larger pore sizes, SWNTs and MWNTs are expected to be more efficient in adsorption of large size molecules. Removal of surface oxygen-containing functional groups from the SWNT enhanced adsorption of AOCs.

Inhalation of formaldehyde and xylene induces apoptotic cell death in the lung tissue.

The aim of this study was to determine the localization and number of apoptotic cells in lung tissue and bronchus-associated lymphoid tissue (BALT) of newborns, young, and adult rats exposed to formaldehyde (6 ppm) or technical xylene (300 ppm) for 6 weeks (8 h/day). A total of 27 female Sprague-Dawley rats were used. Apoptotic cells were mainly localized around the bronchus and bronchioles and relatively less frequently on the walls of alveoli and interalveolar septa both in control and experimental groups. In the BALT, reactive cells were localized in the area under the epithelium and distributed homogenously within the lymphoid follicles. The numbers of apoptotic cells in the lung tissue including the BALT were significantly higher in young and adult rats exposed to formaldehyde and xylene than those detected in control groups.

The right method for midline laparotomy: what is the best choice for wound healing?

BACKGROUND: The techniques used for midline laparotomy affect healing in surgical wounds, but the relationship between cold scalpel and haemostatic methods (harmonic scalpel, diathermy) regarding wound healing remains unclear. There are also limited studies concerning the effects of harmonic scalpel on abdominal fascia. This study was aimed at comparing myofascial wound healing following laparotomy incision on fascia by cold scalpel (CS), harmonic scalpel (HS), and diathermy in terms of hydroxyproline content, inflammatory changes and tensile strength. MATERIALS AND METHODS: Twenty-seven male Wistar albino rats underwent midline laparotomy either with cold scalpel (CS), harmonic scalpel (HS) or diathermy. Fascia incisions were closed with continuous 4/0 polypropylene and skin incisions were closed with interrupted 4/0 polypropylene stitches. On the 7th postoperative day, the abdominal walls of the rats were tested for tensile strength. In addition, each abdominal fascia was evaluated for inflammation scores and hydroxyproline levels. RESULTS: HS caused less inflammation and necrosis in abdominal fascia compared to the diathermy group (p < 0.01 and p < 0.05, respectively), whereas the CS group showed no difference in inflammation scores, but had significantly lower necrosis scores than the HS and diathermy groups (p < 0.05 and p < 0.001, respectively). Hydroxyproline content of the fascia did not differ among groups, while the tensile strength of the wound was obviously higher in the CS group (p < 0.001). CONCLUSION: HS causes less inflammatory reaction and necrosis than diathermy, but more necrosis than CS. Fascia incisions with CS gains tensile strength faster than in other groups. HS appears to cause less tissue injury than diathermy and also has comparable results for wound healing. Further clinical studies on the impact of HS in fascia incisions are needed.

Genetic verification of the role of CCK-AR in pancreatic proliferation and blood glucose and insulin regulation using a congenic rat carrying CCK-AR null allele.

A number of studies have indicated that cholecystokinin type A receptor (CCK-AR) plays a crucial role in postnatal pancreatic proliferation and blood glucose regulation through stimulating insulin secretion. The Otsuka Long-Evans Tokushima Fatty (OLETF) rat has been shown to possess poor pancreatic proliferation (PPP) capability after pancreatectomy (Px). Here we have constructed a congenic strain which introgressed an OLETF-derived 18.5 cM genomic fragment identified in our previous quantitative trait locus (QTL) analysis as a locus responsible for PPP into normoglycemic F344 genetic background The introgressed region includes CCK-AR null mutation. After Px, the congenic rat showed weak pancreatic proliferation equivalent to that of the OLETF rat. Furthermore, post-surgery non-fasting blood glucose levels for the congenic rats are significantly higher in comparison with the F344 rats. At 28 days after Px, the congenic rats also showed lower blood insulin levels than the F344 rats. These results further provide the genetic evidence that 1) CCK-AR is essential for pancreatic regeneration; 2) impaired pancreatic proliferation mediates the development of hyperglycemia.

Genetic evidence for obesity loci involved in the regulation of body fat distribution in obese type 2 diabetes rat, OLETF.

The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese type 2 diabetes in human. Obesity is essential for the onset of type 2 diabetes in this rat. Our present investigation was designed to identify quantitative trait loci (QTLs) contributing to obesity by performing a whole-genome search using 214 F(2) intercross progeny between OLETF and F344 rats. We have identified six QTLs responsible for adiposity indices of fat pads on rat chromosomes 2 (Obs1 for mesenteric fat), 4 (Obs2 for retroperitoneal fat), 8 (Obs3 for mesenteric fat), 9 (Obs4 for retroperitoneal fat), and 14 (Obs5 and Obs6 for retroperitoneal fat), demonstrating that the adiposity indices of individual fat pads were under the control of different genes. As expected, the OLETF allele corresponds to increased adiposity indices for all QTLs, except for Obs3, in which the F344 allele leads to an increase in the index.

Induction of apoptosis and inhibition of DNA topoisomerase-I in K-562 cells by a marine microalgal polysaccharide.

We have previously purified an extracellular polysaccharide, D-galactan sulfate associated with L(+)-lactic acid, produced from a marine microalga Dinoflagellate Gymnodinium sp. A3 (GA3). The GA3 polysaccharide, irrespective of presence or absence of lactic acid, exhibited significant cytotoxicity, which is based on an induction of apoptotic cell death, toward human myeloid leukemia K562 cells. Furthermore, we found that the GA3 polysaccharide with or without lactic acid possesses an inhibitory effect on topoisomerase-I (topo-I). The potent cytotoxic effect of GA3 polysaccharide may result from its inhibitory effect on topo-I, because the topo-I inhibition is known to trigger apoptotic cell death.

Bradykinin regulation of salt transport across mouse inner medullary collecting duct epithelium involves activation of a Ca(2+)-dependent Cl(-) conductance.

The mechanism by which bradykinin regulates renal epithelial salt transport has been investigated using a mouse inner medullary renal collecting duct cell-line mIMCD-K2. Using fura-2 loaded mIMCD-K2 cells bradykinin (100 nM) has been shown to induce a transient increase in intracellular Ca(2+) via activation of bradykinin B2 receptors localized to both the apical and basolateral epithelial cell surfaces. In mIMCD-K2 epithelial cell-layers clamped in Ussing chambers, 100 nM bradykinin via apical and basolateral bradykinin B2 receptors stimulated a transient increase in inward short-circuit current (I:(sc)) of similar duration to the increase in intracellular Ca(2+). Replacements of the bathing solution Na(+) by the impermeant cation, N-methyl-D-glucamine and of Cl(-) and HCO(3)(-) by the impermeant anion gluconate at either the apical (no reduction) or basal bathing solutions (abolition of the response) are consistent with the bradykinin-stimulated increase in inward I:(sc) resulting from basal to apical Cl(-) (anion) secretion. Using the slow whole cell configuration of the patch-clamp technique, bradykinin was shown to activate a transient Cl(-) selective whole cell current which showed time-dependent activation at positive membrane potentials and time-dependent inactivation at negative membrane potentials. These currents were distinct from those activated by forskolin (CFTR), but identical to those activated by exogenous ATP and are therefore consistent with bradykinin activation of a Ca(2+)-dependent Cl(-) conductance. The molecular identity of the Ca(2+)-dependent Cl(-) conductance has been investigated by an RT - PCR approach. Expression of an mRNA transcript with 96% identity to mCLCA1/2 was confirmed, however an additional but distinct mRNA transcript with only 81% of the identity to mCLCA1/2 was identified.

Chemoprevention of DMBA-induced mammary gland carcinogenesis--preventive effects of free oxygen radical scavengers.

This study was carried out to determine the effects of Mangan-Desferrioxamine (Mn-DFX) and Verapamil (VRP) in 7,12-dimethyl-benz-[a]anthracene (DMBA) induced mammary carcinoma. 70 Spraque Dawley rats were divided into four groups; as DMBA alone, DMBA + Mn-DFX, DMBA + VRP, and control. Incidence, multiplicity, and size of tumors were evaluated in addition to analyses of DNA ploidy status and proliferation index by flow cytometric technique. The results revealed that Mn-DFX and VRP caused significant decreases in tumor incidence (p < 0.02 and p < 0.05), multiplicity (p < 0.02 and p < 0.05), size (p < 0.05 and p < 0.02), and proliferation index (p < 0.05 and p < 0.02) without any toxic effect. The appearance of first tumors in Mn-DFX and VRP groups was also later than in the DMBA alone group (10th and 11th week versus 6th week). In conclusion, Mn-DFX and VRP have offered prevention in experimental mammary carcinogenesis. These agents caused slower tumor growth, though they could not achieve a complete prevention.

Homophilic synaptic target recognition mediated by immunoglobulin-like cell adhesion molecule Fasciclin III.

We demonstrate that the cell adhesion molecule Fasciclin III (FAS3) mediates synaptic target recognition through homophilic interaction. FAS3 is expressed by the RP3 motoneuron and its target muscles during synaptic target recognition. The RP3 growth cone can form synapses on muscles that ectopically express FAS3. This mistargeting is dependent on FAS3 expression in the motoneurons. In addition, when the FAS3-negative aCC and SNa motoneuron growth cones ectopically express FAS3, they gain the ability to recognize FAS3-expressing muscles as alternative targets. We propose that homophilic synaptic target recognition serves as a basic mechanism of neural network formation.

Targeted gene expression without a tissue-specific promoter: creating mosaic embryos using laser-induced single-cell heat shock.

We have developed a method to target gene expression in the Drosophila embryo to a specific cell without having a promoter that directs expression in that particular cell. Using a digitally enhanced imaging system to identify single cells within the living embryo, we apply a heat shock to each cell individually by using a laser microbeam. A 1- to 2-min laser treatment is sufficient to induce a heat-shock response but is not lethal to the heat-shocked cells. Induction of heat shock was measured in a variety of cell types, including neurons and somatic muscles, by the expression of beta-galactosidase from an hsp26-lacZ reporter construct or by expression of a UAS target gene after induction of hsGAL4. We discuss the applicability of this technique to ectopic gene expression studies, lineage tracing, gene inactivation studies, and studies of cells in vitro. Laser heat shock is a versatile technique that can be adapted for use in a variety of research organisms and is useful for any studies in which it is desirable to express a given gene in only a distinct cell or clone of cells, either transiently or constitutively, at a time point of choice.

Toll, a muscle cell surface molecule, locally inhibits synaptic initiation of the RP3 motoneuron growth cone in Drosophila.

Toll, a transmembrane molecule with extracellular leucine-rich repeats, is dynamically expressed by the Drosophila embryonic musculature. Growth cones of RP3 and other motoneurons normally grow past Toll-positive muscle cells and innervate more distal muscle cells, which have down-regulated their Toll expression. In this study, we show that reciprocal genetic manipulations of Toll proteins can produce reciprocal RP3 phenotypes. In Toll null mutants, the RP3 growth cone sometimes innervates incorrect muscle cells, including those that are normally Toll-positive. In contrast, heterochronic misexpression of Toll in the musculature leads to the same growth cone reaching its correct target region but delaying synaptic initiation. We propose that Toll acts locally to inhibit synaptogenesis of specific motoneuron growth cones and that both temporal and spatial control of Toll expression is crucial for its role in development.

The prophylactic effects of superoxide dismutase, catalase, desferrioxamine, verapamil and disulfiram in experimental colitis.

In our study we have tried to compare the prophylactic effects of superoxide dismutase (SOD), SOD+catalase (CAT), desferrioxamine, verapamil and disulfiram, which are all free oxygen radical (FOR) scavengers, in an animal model of experimental acetic acid colitis. The control group had 58.3% colonic ulcers. The SOD group (48.6% ulcerated area) and the SOD+CAT group (47.2% ulcerated area) did not show a significant difference when compared with the control group. The values were significantly different in all other groups when compared with the control group (desferrioxamine (29.2% ulcerated area): P < 0.01; verapamil (10.4% ulcerated area) and disulfiram (7.6% ulcerated area): P < 0.001. In conclusion, desferrioxamine, verapamil and disulfiram have been found to be effective as prophylactics in experimental acetic acid colitis.