Changes of antioxidant enzymes in the kidney after cardiac arrest in the rat model.

Globally, cardiac arrest (CA) is a leading cause of death and disability. Asphyxial CA (ACA)-induced kidney damage is a crucial factor in reducing the survival rate. The purpose of this study was to investigate the role of antioxidant enzymes in histopathological renal damage in an ACA rat model at different time points. A total of 88 rats were divided into five groups and exposed to ACA except for the sham group. To evaluate glomerular function and oxidative stress, serum levels of blood urea nitrogen (BUN) and creatinine (Crtn) and malondialdehyde (MDA) levels in renal tissues were measured. To determine histopathological damage, hematoxylin and eosin staining, periodic acid-Schiff staining, and Masson's trichrome staining were performed. Expression levels of antioxidant enzymes including superoxide dismutase-1 (SOD-1), superoxide dismutase-2 (SOD-2), catalase (CAT), and glutathione peroxidase (GPx) were measured by immunohistochemistry (IHC). Survival rate of the experimental rats was reduced to 80% at 6 h, 55% at 12 h, 42.9% at 1 day, and 33% at 2 days after return of spontaneous circulation. Levels of BUN, Crtn, and MDA started to increase significantly in the early period of CA induction. Renal histopathological damage increased markedly from 6 h until two days post-CA. Additionally, expression levels of antioxidant enzymes were significantly decreased at 6 h, 12 h, 1 day, and 2 days after CA. CA-induced oxidative stress and decreased levels of antioxidant enzymes (SOD-1, SOD-2, CAT, GPx) from 6 h to two days could be possible mediators of severe renal tissue damage and increased mortality rate.

Changes of antioxidant enzymes in the kidney after cardiac arrest in the rat model

Globally, cardiac arrest (CA) is a leading cause of death and disability. Asphyxial CA (ACA)-induced kidney damage is a crucial factor in reducing the survival rate. The purpose of this study was to investigate the role of antioxidant enzymes in histopathological renal damage in an ACA rat model at different time points. A total of 88 rats were divided into five groups and exposed to ACA except for the sham group. To evaluate glomerular function and oxidative stress, serum levels of blood urea nitrogen (BUN) and creatinine (Crtn) and malondialdehyde (MDA) levels in renal tissues were measured. To determine histopathological damage, hematoxylin and eosin staining, periodic acid-Schiff staining, and Masson's trichrome staining were performed. Expression levels of antioxidant enzymes including superoxide dismutase-1 (SOD-1), superoxide dismutase-2 (SOD-2), catalase (CAT), and glutathione peroxidase (GPx) were measured by immunohistochemistry (IHC). Survival rate of the experimental rats was reduced to 80% at 6 h, 55% at 12 h, 42.9% at 1 day, and 33% at 2 days after return of spontaneous circulation. Levels of BUN, Crtn, and MDA started to increase significantly in the early period of CA induction. Renal histopathological damage increased markedly from 6 h until two days post-CA. Additionally, expression levels of antioxidant enzymes were significantly decreased at 6 h, 12 h, 1 day, and 2 days after CA. CA-induced oxidative stress and decreased levels of antioxidant enzymes (SOD-1, SOD-2, CAT, GPx) from 6 h to two days could be possible mediators of severe renal tissue damage and increased mortality rate.

Conservation implications of the genetic diversity of Gymnospermium microrrhynchum in Korea.

Gymnospermium microrrhynchum (Berberidaceae) is an ephemeral perennial herb with a limited distributional range in the Baekdudaegan mountain areas of the Korean Peninsula, and is designated a rare plant by the Korean Forest Service. Information about its genetic variation and structure is important for developing successful conservation strategies. To investigate the genetic variation within and among seven G. microrrhynchum populations, random amplified polymorphic DNA data were obtained for 207 individuals. The populations exhibited relatively low genetic diversity: the percentage of polymorphic bands (PPB) ranged from 32.1 to 66.7% (mean = 51.4%) and Nei's gene diversity (HE) ranged from 0.116 to 0.248 (mean = 0.188). However, genetic diversity at the species level was relatively high (PPB = 98.7%, HE = 0.349). An analysis of molecular variance revealed high differentiation among populations (ΦST = 0.6818), but the low gene flow value (Nm = 0.117) suggests a low level of gene exchange occurs among populations. Principal coordinates analysis revealed that individuals were separated according to population. The high level of genetic differentiation and restricted gene flow among G. microrrhynchum populations, which resulted from their isolation in alpine areas after the Ice Age, indicates that it is essential to protect and manage all populations, rather than focus on specific populations, in order to maintain the genetic diversity of this species.

Effect of peracetic acid on biofilms formed by Staphylococcus aureus and Listeria monocytogenes isolated from dairy plants.

This research investigated the removal of adherent cells of 4 strains of Staphylococcus aureus and 1 Listeria monocytogenes strain (previously isolated from dairy plants) from polystyrene microtiter plates using peracetic acid (PAA, 0.5%) for 15, 30, 60, and 120 s, and the inactivation of biofilms formed by those strains on stainless steel coupons using the same treatment times. In the microtiter plates, PAA removed all S. aureus at 15 s compared with control (no PAA treatment). However, L. monocytogenes biofilm was not affected by any PAA treatment. On the stainless steel surface, epifluorescence microscopy using LIVE/DEAD staining (BacLight, Molecular Probes/Thermo Fisher Scientific, Eugene, OR) showed that all strains were damaged within 15 s, with almost 100% of cells inactivated after 30 s. Results of this trial indicate that, although PAA was able to inactivate both S. aureus and L. monocytogenes monospecies biofilms on stainless steel, it was only able to remove adherent cells of S. aureus from polystyrene microplates. The correct use of PAA is critical for eliminating biofilms formed by S. aureus strains found in dairy plants, although further studies are necessary to determine the optimal PAA treatment for removing biofilms of L. monocytogenes.

Association of LTßR gene polymorphisms with prostate volume in benign prostatic hyperplasia in the Korean population.

The lymphotoxin-ß receptor (LTßR) gene is involved in autoimmune disease and inflammatory disorder development, but the relationship between LTßR and benign prostatic hyperplasia (BPH) is unclear. In total, 222 with BPH were examined for 3 single nucleotide polymorphisms [rs3759333 (-1387C/T), rs3759334 (-1326A/G), and rs2364480 (Ala172Ala)] located in the promoter and coding regions of LTßR using direct sequencing. The genotype distributions of rs3759334 were associated with prostate volume larger than 40 g. There were significant differences between the small (<40 g) and large (≥40 g) group subjects [codominant 1 model: odds ratio (OR) = 4.65, 95% confidence interval (CI) = 1.95-11.09, P = 0.001; dominant model: OR = 4.91, 95%CI = 2.07-11.63, P = 0.0002; log-additive model: OR = 4.81, 95%CI = 2.05-11.24, P = 0.0001]. The allele distributions of rs3759334 were significantly associated with BPH (OR = 4.87, 95%CI = 2.16-10.99, P = 0.0001). The distribution of rs2364480 was significantly different between groups (codominant 1 model: OR = 2.17, 95%CI = 1.11-4.26, P = 0.028; dominant model: OR = 2.16, 95%CI = 1.13-4.12, P = 0.019; log-additive model: OR = 1.86, 95%CI = 1.07-3.24, P = 0.027). The allele distribution of rs2364480 was significantly associated with BPH (OR = 1.88, 95%CI = 1.08-3.30, P = 0.027). We found that LTßR polymorphisms caused severe BPH. Thus, LTßR may contribute to the risk of BPH development.

Biofilm-producing ability of Staphylococcus aureus isolates from Brazilian dairy farms.

This study aimed to investigate the in silico biofilm production ability of Staphylococcus aureus strains isolated from milking parlor environments on dairy farms from São Paulo, Brazil. The Staph. aureus isolates were obtained from 849 samples collected on dairy farms, as follows: milk from individual cows with subclinical mastitis or history of the disease (n=220); milk from bulk tank (n=120); surfaces of milking machines and utensils (n=389); and milk handlers (n=120). Thirty-one Staph. aureus isolates were obtained and categorized as pulsotypes by pulsed-field gel electrophoresis and submitted to assays for biofilm formation on polystyrene, stainless steel, rubber, and silicone surfaces. Fourteen (45.2%) pulsotypes were considered producers of biofilm on the polystyrene microplate assay, whereas 13 (41.9%) and 12 (38.7%) pulsotypes were biofilm producers on stainless steel and rubber, respectively. None of the pulsotypes evaluated produced biofilms on silicone. Approximately 45% of Staph. aureus pulsotypes isolated from different sources on dairy farms showed the ability to produce biofilms in at least one assay, indicating possible persistence of this pathogen in the milking environment. The potential involvement of Staph. aureus in subclinical mastitis cases and its occurrence in milk for human consumption emphasize the need to improve hygiene practices to prevent biofilm formation on the farms studied.

Characterization of Staphylococcus aureus isolates in milk and the milking environment from small-scale dairy farms of São Paulo, Brazil, using pulsed-field gel electrophoresis.

This research aimed to evaluate the occurrence of Staphylococcus aureus isolates in milk and in the milking environment of 10 small-scale farms (<400 L/d) located in the regions of Franca and Ribeirão Preto, state of São Paulo, Brazil. Two-hundred twenty samples of milk were collected from individual cows, along with 120 samples from bulk tank milk, 389 samples from milking equipment and utensils (teat cups, buckets, and sieves), and 120 samples from milkers' hands. Fifty-six Staph. aureus strains were isolated from 849 analyzed samples (6.6%): 12 (5.5%) from milk samples of individual cows, 26 (21.7%) from samples of bulk tank milk, 14 (3.6%) from samples collected from equipment and utensils, and 4 (3.3%) from samples from milkers' hands. Pulsed-field gel electrophoresis typing of the 56 Staph. aureus isolates by SmaI restriction enzyme resulted in 31 profiles (pulsotypes) arranged in 12 major clusters. Results of this study indicate a low incidence, but wide distribution of Staph. aureus strains isolated from raw milk collected from individual cows and surfaces of milkers' hands and milking equipment in the small-scale dairy farms evaluated. However, the high percentage of bulk milk samples found with Staph. aureus is of public health concern because raw, unprocessed milk is regularly consumed by the Brazilian population.

Change of telomerase activity in peripheral whole blood of head and neck squamous cell carcinoma patients before and after surgery: a pilot study.

BACKGROUND The purpose of this study was to evaluate telomerase activity in peripheral whole blood from head and neck squamous cell carcinoma (HNSCC) patients as a biomarker for diagnosis of HNSCC or detection of recurrence during follow-up. MATERIALS AND METHODS Telomerase activity was measured from peripheral whole blood extracts by telomerase repeat amplification protocol (TRAP) in HNSCC patients before and after surgery and in a control group. Sixty-two HNSCC patients and 42 control subjects were included. RESULTS Telomerase activity was found in 41 out of 62 (66.1%) HNSCC patients before surgery and in 8 out of 42 (19.0%) controls (p<0.001). Among 41 HNSCC patients who showed positive telomerase activity before surgery, 32 (78.1%) showed a conversion of telomerase activity to negative after surgery. In follow-up, 6 out of 8 (75%) showed conversion of telomerase activity from negative to positive after recurrence. Telomerase activity was changed to negative in 4 out of 6 (66%) recurred patients with positive telomerase activity after second surgery. CONCLUSION The telomerase activity in peripheral whole blood extracts of HNSCC patients might be a useful biomarker for detecting recurrence after treatment. Further study with larger sample size using a more sensitive detection method of telomerase activity is necessary to verify these results.

Standardization of anti-lethal toxin potency test of antivenoms prepared from two different Agkistrodon halys venoms

In Korea, antivenoms for the treatment of patients bitten by venomous snakes have been imported from Japan or China. Although there is cross-reactivity between these antibodies and venoms from snakes indigenous to Korea (e.g. Agkistrodon genus), protection is not optimal. Antivenoms specifically prepared to neutralize Korean snake venoms could be more effective, with fewer side effects. To this end, we established an infrastructure to develop national standards and created a standardized method to evaluate the efficacy of two horse-derived antivenoms using mouse lethal toxin test. Additionally, we determined the antivenoms neutralizing activity against lethal doses (LD50) of Agkistrodon halys (from Japan) and Jiangzhe Agkistrodon halys (from China) venoms. We also performed cross-neutralization tests using probit analysis on each pairing of venom and antivenom in order to check the possibility of using Jiangzhe A. halys venom as a substitute for A. halys venom, the current standard. Slope of A. halys venom with A. halys antivenom was 10.2 and that of A. halys venom with Jiangzhe A. halys antivenom was 9.6. However, Slope of Jiangzhe A. halys venom with A. halys antivenom was 4.7 while that of Jiangzhe A. halys venom with Jiangzhe A. halys antivenom was 11.5. Therefore, the significant difference in slope patterns suggests that Jiangzhe A. halys venom cannot be used as a substitute for the standard venom to test the anti-lethal toxin activity of antivenoms (p<0.05).(AU)

First Report of Chino del Tomate and Pepper Hausteco Geminivurses in Greenhouse-Grown Tomato in Sonora, Mexico.

Tomato plants grown in commercial greenhouses in Sonora, Mexico, developed either yellow mosaic, leaf curling, and stunting (phenotype 1; 20 to 35%) or chlorosis and a feathery appearance of leaves (phenotype 2; 15 to 25%) in December 1997 and again in October 1998. Polymerase chain reaction (PCR) with diagnostic primers (prAV324 and prAC889) that amplify an approximately 576-bp fragment of the coat protein (CP) gene of whitefly-transmitted geminiviruses indicated the presence of a begomovirus. Biolistic inoculation of tomato seedlings with RNase-treated extracts of three symptomatic tomato samples, each, resulted in reproduction of disease symptoms. With PCR primers prAV2644 and prAC1154 (1), the entire CP (approximately 776 bp) and its flanking sequences were amplified from extracts of symptomatic tomato, and amplicons were cloned and sequenced. Comparisons of a minimum of three CP gene sequences from each phenotype revealed the presence of at least two begomoviruses. The phenotype 1-associated CP gene shared 92.6, 86.2, and 85.3% identity with the CP sequence of chino del tomate (CdTV) [AF106936], tomato mottle, and abutilon mosaic geminiviruses, respectively. The CP sequence associated with phenotype 2 was 94.6, 77.1, and 77.1% identical to pepper hausteco (PHV) [X70418], bean golden mosaic-Guatemala (BGMV-GU), and Texas pepper (TPV) gem-iniviruses, respectively. Previously, CdTV was reported from tomato in Chiapas, Morelos, Sinaloa, and Tamaulipas, Mexico, while PHV has been identified in Guanajuato, Quintana Roo, Sinaloa, and Tamaulipas, Mexico, and in Texas (2). This is the first report of CdTV-like (>92% identity) and PHV-like (>94% identity) geminiviruses associated with greenhouse-grown tomatoes in Sonora, Mexico. References: (1) A. M. Idris and J. K. Brown. Phytopathology 88:648, 1998; (2) I. Torres-Pacheco et al. Phytopathology 86:1186, 1996.

Effects of the K+ channel blockers paspalitrem-C and paxilline on mammalian smooth muscle.

The tremorgenic alkaloids, paxilline and paspalitrem-C (0.1-10 microM), increased the spontaneous contractility of guinea-pig and rat urinary bladder, and rat duodenum, and induced tension in guinea-pig trachea. These effects are ascribed to blockade of high-conductance, Ca(2+)-activated K+ (BKCa) channels. Paxilline potentiated the charybdotoxin-induced stimulation of guinea-pig detrusor muscle; this is consistent with the alkaloid's ability to allosterically enhance the binding of charybdotoxin to smooth muscle membranes (Knaus et al., 1994). Paspalitrem-C and paxilline did not affect the myogenic activity of isolated portal vein from guinea-pig, which is insensitive to charybdotoxin, or of that from rat which is stimulated by charybdotoxin. Paxilline and paspalitrem-C also differed from charybdotoxin in that the alkaloids did not consistently elicit tension in guinea-pig aortic rings. These discrepancies are attributed to differences in relative potency, sites and/or mechanisms of action of the indole alkaloids vs. peptidyl blockers of the BKCa channel.

Effects of Brazilin on glucose oxidation, lipogenesis and therein involved enzymes in adipose tissues from diabetic KK-mice.

In order to address the hypoglycemic mechanism of brazilin, effects on glucose metabolism in epididymal adipose tissue from diabetic KK-mice were investigated. Brazilin remarkably lowered non fasting plasma glucose level without any changes in plasma insulin level. Brazilin significantly increased the rate of glucose oxidation and lipogenesis only in the presence of insulin. Activities of glucose-6 phosphate dehydrogenase and fatty acid synthetase, involved in glucose oxidation and lipogenesis respectively, were significantly increased. These results suggest that brazilin might exert hypoglycemic action in insulin resistance state by, at least in part, regulating the enzymatic reaction process involved in glucose metabolism.

Hermansky-Pudlak syndrome: case report and clinicopathologic review.

The Hermansky-Pudlak syndrome is an autosomal recessive disorder consisting of the triad of albinism, a bleeding diathesis, and ceroid deposition within the reticuloendothelial system. In this study of a patient with Hermansky-Pudlak syndrome, we demonstrate the presence of ceroid within dermal macrophages. Electron microscopic studies suggest that melanosomes may be a substrate for the formation of ceroid in the skin. A review of the clinical and pathophysiologic features of this disorder is presented.