Progressive esophageal carcinoma can infiltrate the surrounding tissues with subsequent development of a fistula, most commonly between the esophagus and the respiratory tract. The endoscopic placement of covered self-expanding metallic stents (SEMS) is the treatment of choice for malignant esophageal fistulas and should be performed immediately, as a fistula formation represents a potential life-threatening complication. We report the case of a 64-year-old male diagnosed with esophageal carcinoma, who had a 20Fr surgical gastrostomy tube inserted before chemo- and radiotherapy and was referred to our department for complete dysphagia, cough after swallowing and fever. The attempt to insert a SEMS using the classic endoscopic procedure failed. Then, a fully covered stent was inserted, as the 0.035" guide wire was passed through stenosis retrogradely by using an Olympus Exera II GIF-N180 (4.9 mm in diameter endoscope) via surgical gastrostomy, with a good outcome for the patient. The retrograde approach via gastrostomy under endoscopic/fluoroscopic guidance with the placement of a fully covered SEMS proved to be the technique of choice, in a patient with malignant esophageal fistula in whom other methods of treatment were not feasible.

We observed a case of adult T-cell leukemia (ATL) with proliferative synovitis. Culture supernatants from ATL cells (ATL-SN) obtained from the peripheral blood constitutively produced an interleukin-1 (IL-1)-like factor in vitro, as shown by the growth inhibition factor (GIF) assay using the A375 melanoma cell line and the lymphocyte activating factor (LAF) assay using C3H/HeJ thymocytes. Neutralization studies indicated that polyclonal antibodies against IL-1 beta blocked most (80%) of the activity in ATL-SN. In addition, increased amounts of IL-1 beta mRNA were found in the ATL cells by dot blot analysis. Sephacryl S-200 chromatography showed that the molecular weight of this factor was approximately 17.5 kDa, and Western blot analysis revealed that this factor reacted with polyclonal anti-IL-1 beta antibody under the reduced condition. The isoelectric point was 7.5. Furthermore, ATL-SN showed significant activity in promoting the growth of synovial cells in parallel with IL-1 activity. These data suggest that the constitutive production of this IL-1 beta-like factor might be responsible for proliferative synovitis in this case.

Objective: To evaluate robot-assisted system YunSRobot to perform oesophagogastroduodenoscopy(EGD) in simulation model and human volunteers. Methods: The YunSRobot was invented for soft endoscopy by our team. In this trial, gastroenterologists operated the double handles of YunSRobot to perform EGD endoscopy with Olympus GIF-H 260 in simulation model Lm103 of Koken Japan and human volunteers. The operating time, lumenal anatomic exposure, man-machine interaction and other parameters were recorded and compared with manual endoscopy. Results: In the endoscopy on model, each of four doctors performed 5 procedures of EGD by YunSRobot and traditional manual endoscopy, respectively. The average time of one procedure was (626.4±120.7) seconds in the robot group and (241.5±24.7) seconds in the manual group, the operating time in robot group was more than that in manual endoscopy group. (t=14.0, P<0.001). However, the robotic manipulation time of the four endoscopists was significantly shortened from the first case to the fifth case with ((783.5±154.8)s, (667.75±85.1)s, (582.0±74.7)s, (555.0±28.9)s, (543.8±29.7)s, F=15.353, P=0.03). In the endoscopy on human, three physicians operated the EGD endoscopy in 21 volunteers, each physician performed seven volunteers by YunSRobot and traditional manual endoscopy respectively. The average time of each procedure in the robot group and the manual endoscopy group was (875.6±179.8) vs (378.8±80.4)s, the operating time in robot group was also more than that in manual endoscopy group (t=10.278, P<0.001). Like the endoscopy on model, the robotic manipulation time by the three operators was significantly reduced from the first case to the seventh cases ((954.7±62.1)s, (936.7±116.9)s, (968.7±227.1)s, (1 008.0±229.4)s, (876.7±110.5)s, (735.0±149.2)s, (649.3±81.0)s, F=3.79, P=0.024). All endoscopic anatomy of EGD were clearly and skillfully observed including esophgus, cardia, fundus, body, gastric angle, antrum, pylorus, first and second parts of duodenum, and papilla in both groups. In all procedures, there were no complications. Conclusions: The soft endoscopy robot YunSRobot is competent to conduct endoscopy of EGD on human. Presently the initial learning time of YunSRobot for EGD is longer than that of experienced manual endoscopy. The learning curve shows that the time of robot manipulation reduces rapidly in the initial procedures. It indicates that YunSRobot system has user-friendly functions and is easy to master.

We report a 63-year-old man with recurrent gastroesophageal junction adenocarcinoma. He underwent esophagogastrectomy in August 2004. After curative operation with Stage III (pT3N1M0), a recurrence was found at the anastomosis site in November 2004. Chemoradiotherapy with S-1 followed by chemotherapy (S-1) was performed from January 2005 to April 2006. Lymphnode metastasis to the left side of the main bronchus appeared in May 2006, and paclitaxel was used until December 2007 when PR was indicated by CT scan and GIF. Now he is receiving CPT-11. During these 3 years and 3 months, his performance status was maintained from 0 to 1.

After the initial report of Marshall and Warren, several publications have also demonstrated the presence of pyloric campylobacter in an elevated percentage of cases of chronic gastritis and peptic ulcer. We present our experience studying 672 patients examined by esophagogastroduodenoscopy after referral to clinical because of upper gastrointestinal symptoms. We used an Olympus GIF-K2 panendoscope taking two biopsies from the gastric antrum. Specimens were stained with hematoxilin-eosin and silver. In our experience these methods have given us the best results in detecting this bacteria. Diagnosis of gastric and duodenal ulcer was made by endoscopic criteria and chronic gastritis diagnosed by histology. Chronic gastritis was categorized into active and inactive based in the invasion of polymorphonuclear into mucosa. We have also quantitated damage of the mucin producing cells as well as the degree of bacterial colonization. We have found pyloric campylobacter in 91.8% of patients suffering from chronic active gastritis, 72.7% of patients with diagnosis of gastric ulcer and 84.2% of cases of duodenal ulcer. Our results agree with the ones reported by other authors. We conclude that pyloric campylobacter is not an opportunist microorganism within the stomach because it produces inflammatory changes as well as damage of the mucin producing cells. Its presence may play some role in the etiology of gastritis and peptic ulcer disease.

The ability of GIFs to generate emotionality in social media marketing strategies is analyzed. The aim of this work is to show how neuroscience research techniques can be integrated into the analysis of emotions, improving the results and helping to guide actions in social networks. This research is structured in two phases: an experimental study using automated biometric analysis (facial coding, GSR and eye tracking) and an analysis of declared feelings in the comments of Instagram users. Explicit valence, type of emotion, length of comment and proportion of emojis are extracted. The results indicate that the explicit measure of emotional valence shows a higher and more positive emotional level than the implicit one. This difference is influenced differently by the engagement and the proportion of emojis in the comment. A further step has been taken in the measurement of user emotionality in social media campaigns, including not only content analysis, but also providing new insights thanks to neuromarketing.

Keywords: GIF; Instagram; consumer neuroscience; digital consumer behavior; emotion; eye tracking; facial coding; neuromarketing; sentiment analysis; skin conductance; social networks.

The performance of a pilot stabilization ponds system was evaluated for the treatment of sanitary landfill leachate, focusing on effluent detoxification. Three serial ponds (anaerobic-P1, facultative-P2, and maturation-P3) were evaluated during two different operational conditions: natural condition (period 1) and 24 hours of artificial aeration in P2 (period 2). The ponds system exhibited removal efficiencies of 53% for soluble chemical oxygen demand (SCOD), 81% for filtered biochemical oxygen demand (FBOD), and 84% for NH4+-N during period 1 and 70% for SCOD, 80% for FBOD, and 96% for>-N during period 2. Most metals (Fe, Cu, Cd, Zn, Cr, Ni, and Pb) and organic compounds in the leachate were significantly reduced in the treatment system, with greater reductions during period 2. Moreover, toxicity tests demonstrated the capacity of the system to reduce the toxicity of the raw leachate (up to 89% toxicity reduction).

The ammonia nitrogen stripping process at open horizontal flow reactors was evaluated and is described in this paper. The ammonia came from landfill leachate. Superficial loads of 650 kgN->.ha-1.day-1 (phase 1), 750 kgN->.ha-1.day-1 (phase 2), and 850 kgN->.ha-1.day-1 (phase 3) were applied with a hydraulic retention time of 51.2, 64.9, and 55.6 days, respectively. The experimental system setup was designed, built, and monitored at EXTRABES (Estação Experimental de Tratamentos Biológicos de Esgotos Sanitários) and consisted of a series of four open horizontal flow reactors fed with in natura leachate. The study was based on a stripping process with no alkaline chemical species addition, nor air flow for ammonium gas drag, and aimed to reduce the landfill leachate ammoniacal nitrogen concentration. The studied process showed a removal efficiency of 99.0% in the first phase, 99.3% in the second, and 99.5% in the third. Regarding the organic matter expressed in terms of CODtotal, the removal efficiency was 69.20% in phase 1, 40.12% in phase 2, and 29.23% in phase 3. It was noted that ammonia removal efficiency was directly proportional to the applied superficial load and the carbonaceous material removal was proportional to the organic matter applied in influent, since in phase 1 the CODtotal concentration removal efficiency was 1.7 times greater than that of phase 2, and 2.3 times that of phase 3. Briefly, the ammonia nitrogen stripping process by open horizontal flow reactor is a promising technological alternative for ammonia nitrogen reduction, mainly at regions with available area and temperature greater than 25°C. It is promising because it has no costs in terms of chemical reagents to raise the pH, nor does it require electric costs.

Established methods used to detect serum antibodies to granulocytes require the isolation of granulocytes. Flow cytometric analysis of granulocytes with monoclonal antibodies eliminates the need for granulocyte isolation. The purpose of this study was to develop a method to evaluate reactions of antibodies to granulocytes without separating granulocytes from other leukocytes. Three screening cell samples for granulocyte antibody detection were prepared from whole-blood samples in which the red blood cells (RBCs) were lysed and remaining leukocytes tested against sera at 4 degrees C. Binding of human alloantibodies to the screening cells was determined by flow cytometric analysis using phycoerythrin-conjugated antibody to human immunoglobulin. Forward and side scatter were used to analyze granulocytes separately from other leukocytes. The assay was validated by testing granulocytes with reference alloantibodies directed to NA1, NA2, 5b, and Mart antigens. Samples from 32 patients were tested, and the results of the assays were compared with the results of testing the samples in a granulocyte immunofluorescence (GIF) assay performed by a reference laboratory. In the whole-blood flow cytometric (WBFC) assay the mean fluorescence intensities of reference antisera with antigen-positive cells, expressed in arbitrary units, were anti-NA1 = 48 to 221, anti-NA2 = 24 to 69, anti-5b = 13 to 57, and anti-Mart = 42 to 72. In contrast, the mean fluorescence intensity of type AB-negative control sera ranged from 3 to 11. Of the 32 patient sera tested, 23 were positive (range = 12 to 56) and 9 were negative (range = 3 to 10). When compared with the results obtained by the reference laboratory, 27 sera were concordant between the WBFC and the GIF assays. Four of the samples were positive in WBFC (range = 11 to 31) and negative in GIF and one sample was negative in WBFC (range = 5 to 6) and positive in GIF. Leukocytes prepared from whole blood after lysis of RBCs can be used in flow cytometric analysis to detect granulocyte alloantibodies. The results of testing for granulocyte antibodies with this assay were similar to results of testing sera in GIF. Further comparative studies are indicated to confirm findings and explain the discordant results.

Two patients were admitted to our Gamete Intrafallopian Transfer (GIF) program, and received and ovulation stimulation protocol with Clomiphene Citrate and human menopausal gonadotropin (hMG). Human chorionic gonadotropin (hCG, 10,000 IU IM) was administered when follicle size achieved 18 mm adn oocyte retrieval was scheduled after 36 hours. The estradiol levels in both patients were 1,127 pg/ml respectively at the time hCG was administered, which demonstrates an excellent response to the stimulation. Follicle aspiration was accomplished by transvaginal ultrasound-guided puncture or laparoscopy, and we were unable to recover oocytes in 17 follicles. Our findings support the existence of the "Empty Follicle Syndrome". This pathology may explain some of the infertility causes of our sterile population.

Serum growth hormone (GH) levels in basal conditions (two samples) and 30, 60, 90, 120, 150 and 180 minutes after oral administration of baclofen (20 mg) were evaluated in 6 healthy subjects and in 6 acromegalic patients. The effect of cimetidine (400 mg i.v.) administrated 45 minutes after baclofen (20 mg by mouth) were evaluated in 9 healthy women. Baclofen was able to significantly rise serum GH levels in normal subjects but no in acromegalic patients. Cimetidine suppress GH increase induced by baclofen. It was concluded that: 1) baclofen, GABAb receptor agonist, stimulate GH secretion by inhibition of GIF secretion or by stimulation of GRF secretion; 2) istamine, through H2 receptors in the hypothalamus, is important to mediate GH release induced by stimulation of GABAb receptors.

Carbon nanospheres were prepared and functionalized with carboxyl acid groups (CNS-CA), then reacted with 3-aminopropyltriethoxysilane to introduce amino groups onto the surface (CNS-NH₂) by post-synthesis grafting. CNS-NH₂ was acidified in order to convert the amino groups (-NH₂) into ammonium moieties (>). Various techniques such as N₂ physisorption, X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, thermogravimetry, X-ray photoelectron spectroscopy, and transmission electron microscopy were used to characterize the nanospheres. The removal of chromium ions from aqueous solution using> was investigated. Factors influencing the uptake of Cr(VI) ions such as solution pH, adsorbent dose, and initial Cr(VI) ion concentration were investigated. Equilibrium adsorption data fitted the Langmuir model very well. The adsorption maximum capacity of Cr(VI) was found to be 52.38 mg/g. The reusability of> results indicated that it can be reused five times successfully without loss of adsorption capacity.

A case of a 13-year-old girl is reported. She swallowed her toothbrush accidentally, the surgeon suggested the endoscopic examination. The teen-age, well-developed girl did not have complaints, there were five hours between the accident, and the starting of the examination. After the pharynx anaesthetization by Lidocaine spray, the endoscopy was performed by a video endoscope of GIF-130 type. The brush was found in the stomach, but its handle went through the pylorus. Injury was not found on the esophagus and stomach mucosa. The patient and her mother were fully informed, and for their request, the removal of the toothbrush was attempted by using grasping forceps and polypectomy snare. It lasted nearly forty minutes and was ensured by the intensive care. The procedure was successful, the patient who tolerated it well, left the hospital after observation, without complaints.

We propose a new metric for long-range transport potential (LRTP), GIF, based on source-receptor analyses and evaluate the LRTP and persistence of a wide variety of chlorinated and brominated organic compounds using GIF and overall persistence (POV), respectively. We calculated GIF and POV using our global 3D dynamic multimedia model (FATE). Physicochemical properties were obtained from quantitative structure-property relationship (QSPR) models. The FATE-QSPR combined model enabled us to systematically investigate the LRTP and persistence of a wide variety of chemical substances. On average, the estimated GIF and POV for chlorinated compounds were larger than those for their brominated counterparts, with the largest and smallest values found for polychlorinated biphenyls and polybrominated dibenzodioxins, respectively. We also compared GIF with four differently defined LRTP metrics and two LRTP metrics obtained from a simple model. The results of our analyses indicate that the LRTP ranks can differ considerably among LRTP metrics, the differences being dependent on the governing environmental processes, relevant physicochemical properties, and multimedia model.

Studies aimed at a comparison of chemical, biomimetic (Gif system GoAgg(III))and enzymatic (CHMO) transformations of natural (+)-10beta,14-dihydroxy-allo-aromadendrane have led to preparation of an eight-member sesquiterpene lactone.

This study aimed to determine the interference of nitrite (>) with soluble chemical oxygen demand (COD) measurements from the effluent of landfill leachate treated by sequencing batch reactors (SBRs). Synthetic wastewater assimilating young and old landfill leachate was used as influent for SBR1 and SBR2, respectively. A mixture of raw wastewater and landfill leachate was used as influent for SBR3. Due to the high ammonia concentration in the influent, different levels of partial nitrification were observed in all the reactors and> was detected in the effluents. Theoretically, when> is present in the effluent, 1 mg/L of> accounts for 1.1 mg/L of COD (i.e., 1.1 mg COD/mg>) due to the oxidation of> to>. It was found that the value of 1.1 mg COD/mg> was 3 times higher than the values obtained in the experiments with SBR1 and SBR3. In addition, the values obtained in the experiments with SBR2 were much higher than 1.1 mg COD/mg>. These results suggest that the theoretical value of 1.1 mg COD/mg> cannot be applied to predict the COD values caused by nitrite in the tested wastewaters. To obtain an accurate measurement of soluble COD in samples that contain nitrite, nitrite should be eliminated before the measurements.

Abstract We evaluated the efficacy of omeprazole (OPZ) for the treatment of upper gastrointestinal (UGI) lesions in rheumatoid arthritis (RA) patients. Fourteen RA patients with H2 receptor antagonist- (H2RA-) resistant UGI lesions (1 stomal, 11 gastric, and 2 esophageal with reflux esophagitis ulcers) were treated with OPZ at 20 mg/day (study A). New untreated UGI lesions (1 stomal and 12 gastric ulcers) were treated with OPZ (study B). Three patients who showed renal dysfunction during H2RA treatment for UGI lesion were treated with OPZ (study C). Nonsteroidal antiinflammatory drugs (NSAIDs) were not discontinued. The stage of each ulcer was determined by gastrointestinal fiberscopy (GIF). In study A, during the first 8 weeks of OPZ treatment, 1 esophageal and 7 gastric ulcer patients were completely cured. Six patients showing partial response were treated further with OPZ for another 8 weeks. During this second period, 1 stomal and 3 gastric ulcer patients were completely cured, and 1 gastric and 1 esophageal ulcer patient showed only partial response. In study B, after an 8-week OPZ treatment, all except 2 patients showed complete healing. One patient developed mild eruption at 4 weeks and was shifted to H2RA. One patient showed complete healing after 4 weeks. No patient in study C showed renal dysfunction with OPZ. Our results suggest that OPZ is an effective treatment for UGI lesions in RA patients using NSAIDs.

A year-long field investigation was carried out in the most heavily polluted bay of Lake Chaohu to assess the temporal exchanges of ammonium nitrogen (>) and soluble reactive phosphorus (SRP) across the sediment-water interface (SWI) and to provide remediation advises. Results showed that the monthly average fluxes of> and SRP were 31.38 and 6.98 mg m-2 d-1, respectively, both of which were higher than those in many other hyper-eutrophic lakes around the world. The exchanges of> and SRP were both closed related to the oxygen penetration. Low oxygen penetration depth and generally negative oxygen uptake rates provoked the dissolution of redox sensitive phosphorus and labile> in the sediment and increased the fluxes. In addition, the generally higher fluxes during late spring to autumn should be noted during the reduction of internal loadings, when applicable techniques should be implemented accordingly to achieve better reduction effects.

Immunization with alum-absorbed ovalbumin (OA) failed to induce IgE antibody responses in SJL mice, while mice pretreated with either cyclophosphamide (CY) or gamma-irradiation prior to immunization transiently formed IgE antibodies. Spleen cells of OA-primed SJL mice formed IgE-suppressive factors upon incubation with OA. In contrast, spleen cells of the CY-treated or irradiated mice formed IgE-potentiating factors. It was found that CY treatment diminished the formation of glycosylation inhibiting factor (GIF) and enhanced the formation of glycosylation enhancing factor (GEF). The results suggest that the enhancement of the IgE antibody response by CY treatment is due not only to elimination of suppressor T cells but also to the activation of GEF-forming lymphocytes.

Growth inhibitory factor (GIF), a member of the metallothionein (MT) family, is also known as MTIII. This protein distinguishes itself from other MT isoforms by exerting an inhibitory effect on cortical neuron growth instead of metal ion buffering. In this work, we cloned MTIII genes from a porcine brain cDNA library. Two species of clones were isolated that vary with respect to one nt in the coding sequence. This discrepancy results in the translation of two MTIII primary structures having a different amino acid at residue 46. Herein, both MTIII cDNAs were constructed into an expression vector and transformed into yeast cells, respectively. The yeast carrying either MTIII gene displayed a similar metal tolerance when cultured in a medium containing metal. The resistance to metal toxicity was attributed to the expression of MTIII gene which was confirmed by RNA and protein analyses. The characteristics of the protein stability, metal binding affinity and ultraviolet absorption spectrum of the yeast produced MTIII are also compared with those of MTII. The comparison reveals that both MTs have similar physical characteristics. Moreover, circular dichroism spectrum of Cd saturated MTIII was analyzed as well. Typical Cys-Cd bands for MTII appear in the spectrum, indicating similar metal-thiol interactions for MTIII to those for other MT isoforms.

OBJECTIVE

To observe the morphology of rabbit corneal nerve regeneration in the grafts after penetrating keratoplasty (PKP) with long-term cryopreserved homogeneous donor material.

METHODS

Both long-term cryopreserved and short-term moist-chamber preserved fresh rabbit corneas were applied in the keratoplasty. By means of glyoxylic-acid induced fluorescence (GIF) and acetylcholine esterase (AChE) staining histochemical techniques, the morphological features of adrenergic and cholinergic nerve regeneration were monitored under light microscope in those grafts from ten days to eight months after surgery.

RESULTS

During various intervals, the morphological changes of neural reinnervation were similar in the two groups, and numerous subepithelial nerves that showed random changes in direction and caliber were apparent. The nerve endings in the epithelium were few.

CONCLUSIONS

The cryopreservative procedures do not affect neural regeneration under light microscope. The depressed tactile sensitivity following the surgery is correlated with nerve endings in the epithelium subnormal in number and with the regeneration of corneal nerves abnormal in distribution and morphology.

The aim of this work was to compare efficiency of a new digital endoscopic system in combination with the NBI technology and routine endoscopic examination of patients with chronic gastritis. 181 of the 831 patients with different forms of chronic gastritis were examined with a GIF-H180 Evis Exera-II apparatus (Olympus, Japan) coupled to the NBI system. The control group comprised 650 patients examined using conventional GIF-Q160 and GIF-Q160 Evis Exera-I fiberscopes (Olympus, Japan). Signs of inflammation, its severity and extent were evaluated. Significant correlation was revealed for the first time between inflammation of gastric mucosa, type of gastritis, endoscopic picture in the NBI regime, and results of histologic studies. Inflammation foci and sites of intestinal metaplasia were found to produce characteristic images in which they had different shades of lilac and deep blue respectively. Distribution of the stained foci corresponded to that of inflammation and/or metaplasia. It is concluded that the use of the NBI technology in routine oesophagogastroduodenoscopy improves diagnosis of intestinal metaplasia to 80-100% (p = 0.000).

Although stratification in deep lakes is well-discussed, few studies pay attention to thermal structure as well as its influences on stratification of biochemical parameters in subtropical lakes in mountainous cities. Here, we studied the depth profile of temperature and biochemical parameters in Longjing Lake, a subtropical reservoir in a mountainous city. Thermal stratification became strong during summer. Biochemical parameters were strongly associated with thermal structure. Stratification started at 2~6 m depth with a substantial decrease in dissolved oxygen, biochemical oxygen demand (BOD₅), chlorophyll a, and pH, corresponding to an increase in total nitrogen, ammonium (<inline-graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="02036-ilm01.gif"/>), nitrite (<inline-graphic xmlns:xlink="http://www.w3.org/1999/xlink" xlink:href="02036-ilm12.gif"/>), total inorganic nitrogen (TIN), total phosphorus, and soluble reactive phosphorus (SRP) with depth; the majority of biochemical parameters showed slight variations from 12 m downward. Our results indicated the stratification of Longjing Lake was stronger and more stable than the stratification of tropical and temperate lakes in lowland cities.

The study was designed to assess the value of a new NBI-based digital videoendoscopic system for the diagnosis of erosive and ulcerative lesions in the stomach and duodenum of 136 patients in comparison with 185 ones examined with the use of conventional fiber-optic and videoendoscopes. Patients of group 1 were examined using a GIF-H180 EVIS Exera-I apparatus ("Olympus", Japan), controls with GIF-Q40 and GIF-Q160 EVIS Exera-II endoscopes ("Olympus", Japan). Endoscopic signs of inflammation, its extent and severity were recorded. NBI endoscopy revealed perifocal inflammatory changes associated with ulcerative lesions in the stomach and duodenum as spots of different tones of blue. This method allowed for the detection of lesions not visualized by conventional techniques. The use of NBI technology for routine endogastroduodenoscopy in patients with erosive and ulcerative lesions in the stomach and duodenum improves visualization of even minor changes in gastric and duodenal mucosa (p < 0.05).