Selective Growth Suppressive Effect of Pravastatin on Senescent Human Lung Fibroblasts.

Various chemical reagents containing inhibitors of mitochondrial activity, antioxidants, nuclear factor-kappa B (NF-kB) inhibitor, mammalian target of rapamycin (mTOR) inhibitor and other clinical therapeutics were screened in order to identify those that selectively decrease the viability of senescent human lung fibroblasts. Cell viability was measured using the CCK-8 assay. The results showed that pravastatin, a drug for hyperlipidemia, decreased the viability of senescent cells but not non-senescent cells. The effect of pravastatin on senescent cells is thought to be due to the inhibition of cell proliferation, rather than cell death. The effect of pravastatin was further investigated using the glucose metabolism assay, which showed that glucose consumption was inhibited both in non-senescent and senescent cells and intracellular nicotinamide adenine dinucleotide (NAD) was decreased in senescent cells. Changes to the mRNA expression levels of senescence-associated genes in response to pravastatin treatment were quantified by real-time-qPCR. There were no significant changes in the relative mRNA expression levels of IL-1ß, p16, p21, and p53 in pravastatin-treated non-senescent cells, whereas the expression of IL-1ß and p16 were increased by pravastatin only in senescent cells. The results of this study suggest that pravastatin does not induce senolysis, but rather selectively inhibits the proliferation of senescent cells and that cellular senescence is enhanced by decreasing intracellular NAD and promoting IL-1ß production.

Motor strategy patterns study of diabetic neuropathic individuals while walking. A wavelet approach.

The aim of this study was to investigate muscle׳s energy patterns and spectral properties of diabetic neuropathic individuals during gait cycle using wavelet approach. Twenty-one diabetic patients diagnosed with peripheral neuropathy, and 21 non-diabetic individuals were assessed during the whole gait cycle. Activation patterns of vastus lateralis, medial gastrocnemius and tibialis anterior were studied by means of bipolar surface EMG. The signal׳s energy and frequency were compared between groups using t-test. The energy was compared in each frequency band (7-542 Hz) using ANOVAs for repeated measures for each group and each muscle. The diabetic individuals displayed lower energies in lower frequency bands for all muscles and higher energies in higher frequency bands for the extensors׳ muscles. They also showed lower total energy of gastrocnemius and a higher total energy of vastus, considering the whole gait cycle. The overall results suggest a change in the neuromuscular strategy of the main extensor muscles of the lower limb of diabetic patients to compensate the ankle extensor deficit to propel the body forward and accomplish the walking task.

Time-frequency analysis methods for detecting effects of diabetic neuropathy.

There have been several research studies on efficient methods for analysis and classification of electromyography (EMG) signals and adoption of wavelet functions, which is a promising approach for determining the spectral distribution of the signal. This study compares distinct time-frequency analysis methods for investigating the EMG activity of the thigh and calf muscles during gait among non-diabetic subjects and diabetic neuropathic patients. It also attempts to verify, by adaptive optimal kernel and discrete wavelet transform, whether there are EMG alterations related to diabetic neuropathy in the lower limb muscles during gait. The results show that diabetics might not keep up with the mechanical demands of walking by changing muscle fibre recruitment strategies, as seen in the control group. Moreover, principal components analysis indicates more alterations in diabetic motor strategies, and we identify that diabetic subjects need other strategies with different muscle energy production and frequencies to carry out their daily activities.

Inter- and intra-tester reliability of clinical measurement to determine medio-lateral patellar position using a pachymeter or visual assessment.

The aim was to investigate inter-tester and intra-tester reliability and parallel reliability between a visual assessment method and a method using a pachymeter for locating the mid-point of the patella in determining the medial/lateral patella orientation. Fifteen asymptomatic subjects were assessed and the mid-point of the patella was determined by both methods on two separate occasions two weeks apart. Inter-tester reliability was obtained by ANOVA and by intraclass correlation coefficient (ICC); intra-tester reliability was obtained by a paired t-test and ICC; and parallel reliability was obtained by Pearson's Correlation and ICC, for the measurement on the first and second evaluations. There was acceptable inter-tester agreement (p=0.490) and reliability for the visual inspection (ICC=0.747) and for the pachymeter (ICC=0.716) at the second evaluation. The inter-tester reliability in the first evaluation was unacceptable (visual ICC=0.604; pachymeter ICC=0.612). Although there was statistical similarity between measurements for the first and second evaluations for all testers, intra-tester reliability was not acceptable for both methods: visual (examiner 1 ICC=0.175; examiner 2 ICC=0.189; examiner 3 ICC=0.155) and pachymeter (examiner 1 ICC=0.214; examiner 2 ICC=0.246; examiner 3 ICC=0.069). Parallel reliability gave a perfect correlation at the first evaluation (r=0.828; p<0.001) and at the second (r=0.756; p<0.001) and reliability was between acceptable and very good (ICC=[0.748-0.813]). Both visual and pachymeter methods provide reliable and similar medial/lateral patella orientation and are reliable between different examiners, but the results between the two assessments at 2 weeks' interval demonstrated an unacceptable reliability.

Role of ankle mobility in foot rollover during gait in individuals with diabetic neuropathy.

BACKGROUND: The purpose of this study was to investigate the ankle range of motion during neuropathic gait and its influence on plantar pressure distribution in two phases during stance: at heel-strike and at push-off. METHODS: Thirty-one adults participated in this study (control group, n=16; diabetic neuropathic group, n=15). Dynamic ankle range of motion (electrogoniometer) and plantar pressures (PEDAR-X system) were acquired synchronously during walking. Plantar pressures were evaluated at rearfoot, midfoot and forefoot during the two phases of stance. General linear model repeated measures analysis of variance was applied to investigate relationships between groups, areas and stance phases. FINDINGS: Diabetic neuropathy patients walked using a smaller ankle range of motion in stance phase and smaller ankle flexion at heel-strike (P=0.0005). Peak pressure and pressure-time integral values were higher in the diabetic group in the midfoot at push-off phase when compared to heel-strike phase. On the other hand, the control group showed similar values of peak pressure in midfoot during both stance phases. INTERPRETATION: The ankle mobility reduction observed could be associated to altered plantar pressure distribution observed in neuropathic subjects. Results demonstrated that midfoot and forefoot play a different role in subjects with neuropathy by receiving higher loads at push-off phase that are probably due to smaller ankle flexion at stance phase. This may explain the higher loads in anterior areas of the foot observed in diabetic neuropathy subjects and confirm an inadequate foot rollover associated to the smaller ankle range of motion at the heel-strike phase.

Antioxidant activity and inhibitory effect of Gamazumi (Viburnum dilatatum THUNB.) on oxidative damage induced by water immersion restraint stress in rats.

The antioxidant activity and antiulcer effects of Gamazumi (Viburnum dilatatum THUNB.) fruit squeezing solution (GSS) were investigated. GSS including no antioxidant additive showed strong antioxidant activity by the XYZ-dish method and the electron spin resonance (ESR) method. GSS showed to have no negative effect on growth of rats for 2 weeks of feeding with free access to GSS. After feeding, the GSS group showed a significant inhibitory effect on gastric ulcer formation by water immersion restraint stress for 6 hours compared with the Water group. Plasma, liver and stomach concentrations of lipid peroxide in the GSS group were reduced rather than the water group. Furthermore, the activities of plasma lactic dehydrogenase, amylase and creatine phosphokinase are ordinarily increased by stress; however these activities in the GSS group decreased to the level in the Control group having no stress. The physiological effects of GSS were similar to, or higher than, those of 0.1% (-)-epigallocatechin gallate (EGCg) solution. These effects of EGCg and GSS were similar to the order of antioxidant activity against hydroxyl radical found by both the XYZ-dish and the ESR methods. It was concluded that after ingestion of GSS, during the period of strong antioxidant activity in the body, it could prevent stress-induced oxidative damage.

Stereo acuity in patients with unilateral macular hole and after unilateral macular hole surgery.

PURPOSE: To investigate stereo acuity levels in patients with unilateral idiopathic macular hole and after surgical intervention. METHODS: In 31 consecutive patients with a unilateral macular hole and 46 consecutive patients who underwent successful unilateral macular hole surgery, complete ocular examinations, including orthoptic examinations and microperimetry using the scanning laser ophthalmoscope, were performed. RESULTS: A significantly positive correlation was found between VA and stereo acuity (r = 0.87, P < 0.01). After successful surgery, stereo acuity also correlated with the presence or absence of absolute and/or relative scotoma, and was best in eyes without scotomata. Patients with unilateral idiopathic macular hole, suppression, and symptom duration of 24 months or longer had no stereoscopic vision. CONCLUSIONS: The results indicated that in patients with unilateral idiopathic macular hole and after surgery, stereo acuity correlated with VA. Patients with unilateral macular hole should be operated upon as early as possible, resulting in better VA and better stereo acuity.

In vitro and in vivo antibacterial activity and pharmacokinetics of SC-002 and its derivative, SC-004: new oral cephalosporins.

SC-002 is a novel oral cephalosporin possessing a unique thiadiazolylethenyl moiety at the 3 position. In the present study, it was the most active against gram-positive bacteria among oral cephalosporins such as cefdinir (CFDN), cefpodoxime, cefditoren and cefaclor (CCL). It was equal to or 16 times more active than CFDN against standard and clinical strains. In particular, against clinical isolates of Morganella morganii and Haemophilus influenzae, SC-002 was 8-64 times more active than CFDN. The antibacterial activity of SC-002 against some beta-lactam-resistant strains was superior to that of CFDN. The in vivo antibacterial activity of SC-004, a pivaloyloxymethyl ester of SC-002, was 1.2-8 times more protective against systemic infections due to Streptococcus pneumoniae, Escherichia coli and Klebsiella pneumoniae than that of CFDN. The therapeutic effects of SC-004 on experimental respiratory tract infections caused by S. pneumoniae or H. influenzae were superior to those of CFDN and CCL. SC-004 showed higher and longer-lasting blood levels and higher urinary excretion in pharmacokinetics in mice.

[Effects of a short-term pulmonary rehabilitation program on patients with chronic respiratory failure due to pulmonary emphysema].

To evaluate the effects of a short-term pulmonary rehabilitation program on dyspnea, exercise capacity, and lung function, 15 patients with chronic respiratory failure due to pulmonary emphysema were enrolled in such a program for 3 weeks as inpatients. The program consisted of pursed lip breathing, diaphragmatic breathing, respiratory muscle stretch gymnastics, and walking with synchronized breathing. Dyspnea as measured with a visual analogue scale at the end of a 6-minute walk before and after the program (49.7 +/- 4.0% to 24.2 +/- 3.8%) decreased significantly (p < 0.01). As a measure of functional exercise capacity, the 6-minute walking distance (226.9 +/- 32.4 m to 292.1 +/- 35.8 m) increased significantly (p < 0.01). As an indicator of maximal exercise capacity, endurance time on an incremental treadmill test did not improve. Spirometric data did not change during the study. Total lung capacity (TLC) (8.44 +/- 0.70 L to 7.58 +/- 0.74 L) and residual volume (RV) (5.13 +/- 0.53 L to 4.28 +/- 0.59 L) decreased significantly (p < 0.01). The findings suggest that this program relieves dyspnea, increases functional exercise capacity, and decreases TLC and RV on patients with chronic respiratory failure due to pulmonary emphysema.

Effectiveness of the anticholinergic agent oxitropium bromide in elderly patients with bronchial asthma.

The short-term and long-term effects of the anticholinergic agent oxitropium bromide (CAS 30286-75-0, OTB) were studied in nine elderly patients with bronchial asthma (8 men and 1 women, age: 69.5 +/- 5.2 years, 3 atopic and 6 (not atopic cases). 2 h after inhalation of 0.8 mg of OTB, forced vital capacity (FVC) had increased from 2.28 +/- 0.22 1 to 2.87 +/- 0.27 1 (p < 0.01), forced expiratory volume in 1 s (FEV1) had increased from 1.22 +/- 0.14 to 1.57 +/- 0.16 1 (p < 0.01), and respiratory resistance (Rrs) had decreased from 5.9 +/- 0.8 cmH2O/l/s to 4.9 +/- 0.7 cmH2O/l/s (p < 0.05). The patients inhaled OTB (0.2 mg t.i.d.) for 6 weeks. Their peak expiratory flow rates, measured each morning and each evening, increased significantly during the first 2 weeks of administration, and the increase was sustained for the entire weeks (p < 0.05). Other pulmonary-function tests were done before drug administration and again at the end of the 6 weeks. By the end of the 6th week of OTB administration, FVC had increased from 2.30 +/- 0.22 1 to 2.99 +/- 0.26 1 (p < 0.01), and FEV1 had increased from 1.25 +/- 0.17 to 1.70 +/- 0.22 1 (p < 0.05). These data indicate that OTB can be useful in the treatment of elderly patients with bronchial asthma.

Expression of E- and P-selectins by vascular endothelial cells in human tonsils.

Selectins play an important role in the adhesion between vascular endothelial cells and leukocytes in immuno-inflammatory events. We investigated the immunohistochemical localization of E- and P-selectins in the human palatine tonsils at light and electron microscopic levels. E-selectin was expressed by high endothelial venules (HEV) in a linear pattern in heavily inflamed tonsils with focal infection and extensive neutrophilic infiltration, and much less in hypertrophic tonsils without focal infection. P-selectin expression was found on HEV to the same extent in both disease states in a dot pattern in the cytoplasm. Immunoelectron microscopically, E- and P-selectins were localized along the luminal plasma membrane and in the perinuclear space in HEV endothelial cells. The present study suggests that increased expression of E- and P-selectin in vascular endothelial cells is a significant feature of inflamed tonsils induced by focal infection. E- and P-selectins may participate in different roles in the inflammatory process of the tonsils.

Survey of natural scrapie in Japan: analysis of RFLP types of the PrP gene and detection of PrPSc mainly in Suffolk sheep.

We examined the brains, spleens and/or lymph nodes of 197 mainly suffolk sheep collected from Hokkaido, and the Tohoku, Kanto and Chubu districts to detect PrPSc and thus to estimate scrapie contamination in Japan. Sixteen sheep in Hokkaido and 2 sheep in other districts that were introduced from Hokkaido were positive for PrPSc. By comparison of the frequencies of the restriction fragment length polymorphism (RFLP) types of these 18 scrapie sheep with 128 healthy sheep, we confirmed the association of specific RFLP types of the PrP gene with natural scrapie. The frequency of RFLP type I in scrapie sheep was significantly higher than that in healthy sheep and those of other types in scrapie sheep. In contrast, the frequencies of type II and VI in healthy sheep were higher than those in scrapie sheep. Therefore, type I sheep seemed to be susceptible but type II and VI sheep seemed to be resistant to natural scrapie in Suffolk sheep in Japan. Furthermore, we investigated the distribution of the RFLP types of the PrP gene in 161 sheep in Japan to learn about the genetic background of the susceptibility to scrapie. There were variations in the distribution of the RFLP types in each district.

Detection of PrPSc in sheep at the preclinical stage of scrapie and its significance for diagnosis of insidious infection.

A field sheep insidiously infected with natural scrapie was diagnosed at the preclinical stage through detection of the core fragment of PrpSc (PrPcore) in peripheral lymph nodes by the biopsy method. Three out of 32 euthanized healthy sheep were found positive for PrPcore in the spleen and lymph nodes. Mice that were inoculated with spleen homogenate of one of these sheep showed clinical signs of scrapie and were positive for PrPcore in their brain samples. These results suggest that the detection of PrPcore is significant for diagnosis in the preclinical phase of scrapie.

Post-synaptic potentiation of vagal contractile responses by neurokinin A.

We have studied the effect of neurokinin A (NKA) on tracheal smooth muscle (TSM) contraction caused by administration of acetylcholine (ACh) intra-arterially (ia) into the tracheal circulation, or bilateral stimulation of the vagus nerves, in 26 mongrel dogs. The tracheal contractile response to 10(-7) mol NKA was 3.12 +/- 0.48 g/cm, 6.3 +/- 0.7%AChmax. Plasma histamine concentration was measured in the right atrium at the peak response following administration of NKA. Plasma histamine concentration was 3.33 +/- 2.05 ng/ml after 10(-7) mol NKA (vs. 1.82 +/- 0.54 ng/ml for control, P > 0.10). A frequency-response curve was generated by electrical stimulation of the caudal ends of cut cervical vagi over the range of frequencies 1-20 Hz (constant 30 V) at 15 s intervals 10 min before and 10 min after administration of 10(-8) mol NKA. Substantial augmentation of tracheal contractile responses to 10(-10)-10(-7) mol ia ACh was obtained 10 min after administration of NKA (P < 0.05). Significant potentiation of tracheal contraction to vagal stimulation for 10-20 Hz was also observed after administration of 10(-8) mol NKA (P < 0.05). We have demonstrated a substantial contractile effect of NKA on canine tracheal smooth muscle that is not related to histamine release from respiratory mast cells. We have also demonstrated that NKA causes augmentation of the parasympathetic tracheal contractile response induced by ia administration of ACh and efferent vagus nerve stimulation. These data suggest that potentiation of vagal contractile response by NKA is related to post-synaptic activation of the parasympathetic nerves.