N-Acetyl-l-Cysteine Attenuates Ischemia/Reperfusion Injury–Induced Allodynia and N-Methyl-d-Aspartate Receptor Activation in Rats

Background

Although reactive oxygen species (ROS) are believed to be involved in pathogenic mechanisms that underlie complex regional pain syndrome type I (CRPS-I), the role of ROS in the central mechanism of CRPS is not fully understood.

Objective

In this study we investigated whether ROS scavenger N-acetyl-l-cysteine (NAC) was capable of attenuating mechanical allodynia and whether pain was decreased through modulating N-methyl-d-aspartate (NMDA) receptor activation in a chronic postischemia pain (CPIP) animal model that mimics the symptoms of CRPS-I.

Methods

Thirty male Sprague-Dawley rats were randomly allocated to 5 different groups: (1) sham rats and CPIP rats treated with (2) vehicle; (3) NAC 30 mg/kg; (4) NAC 100 mg/kg; and (5) NAC 300 mg/kg intraperitoneally at 15 minutes before reperfusion. CPIP was generated after a 3-hour ischemia/reperfusion injury on the hind limb using a tight fitting O-ring. Then, mechanical paw-withdrawal thresholds to von Frey stimuli were assessed before ischemia (baseline), at 4 hours; 1, 3, and 5 days; and 1, 2, 3, and 4 weeks after reperfusion. Another set of 5 animal groups in the same categories was used to determine phosphorylated NMDA receptor 1 subunit (pNR1) immunoreactivity in the ipsilateral L4/6 spinal cord at 3 days after reperfusion.

Results

The sham group showed no significant difference in pain thresholds over 4 weeks. With NAC treatment, the pain thresholds measured after reperfusion increased significantly, and this increase lasted 4 weeks after reperfusion compared with the vehicle group (P < 0.01 on the ipsilateral side and P < 0.05 on the contralateral side). The relative density of pNR1 at 3 days after reperfusion in NAC-treated rats decreased significantly compared with that of the vehicle group (all, P < 0.001). The NAC dose was significantly correlated not only with paw-withdrawal threshold (ρ = 0.979; P < 0.001) but also with the relative density of pNR1 (ρ = –0.875; P < 0.001).

Conclusions

NAC, administered during the pre-reperfusion period, had a long-term antiallodynic effect through the attenuation of NMDA receptor phosphorylation, leading to central sensitization.     

Study of the genetic discrimination between imported and autochthonous cases of malaria in South Korea

There has been a great increase of Plasmodium vivax incidences in the Republic of Korea and the genetic diversity of the parasite became more complex with the rapid dissemination of newly introduced genotypes. Surveillance of imported malaria is very important, but there is no good way to determine imported vs. internal cases. In this study, we characterized imported vivax cases, analyzed the genetic sequence of three imported vivax malaria cases for the merozoite surface protein-1 (MSP-1) and circumsporozoite protein (CSP) genes, and clearly discriminated an imported vivax case that was misdiagnosed as indigenous by genetic analysis. PCR reaction for the merozoite surface protein-1 (MSP-1) and circumsporozoite protein (CSP) genes from three imported vivax cases were amplified and sequenced. The genetic variations were compared with a previously constructed database of South Korean isolates. The imported vivax cases showed various patterns on incubation period before onset. Most cases were from other parts of Asia. The MSP-1 gene sequence analysis of three imported cases showed that the imported cases had completely different sequences from any subtypes from Korean isolates. Case-1 and Case-2 exact match with an Indian isolate, and Case-3 had great similarity with isolates from countries neighboring Indonesia. CSP gene analysis based on the repeat patterns showed similar results that the sequences from the imported cases well matched with the patient's traveled countries and completely discriminated with indigenous cases. AMA-1 gene analysis also supported these results. We were able to clearly distinguish three imported vivax cases from indigenous by using a genetic database of Korean isolates and were able to suspect its origin by genotyping. This study demonstrated the usefulness of genetic survey on imported malaria cases.     

Hypermethylation of promoter regions of the APC1A and p16INK4a genes in relation to prognosis and tumor characteristics in cervical cancer patients

Hypermethylation of the O6-MGMT, p14ARF, p16INK4a, RASSF1A and APC1A genes are unfavourable prognostic markers in colorectal cancer (CRC). We hypothesized that they could be related to prognosis also in cervical cancer. Methylation was studied in DNA extracts from surgical specimens of cancer tissue by novel pyrosequencing methods. In 109 patients (90 squamous cell carcinomas, 19 adenocarcinomas), we found that hypermethylation of the APC1A gene promoter occurred in 8.3% of patients, and of p16INK4a in 1.8%. APC1A hypermethylation was significantly related to more advanced FIGO stage of the tumor (P=0.013), larger tumor diameter (P=0.049) and distant recurrence-free survival (P=0.0007), but not with locoregional recurrence rate, age, HPV status, DNA ploidy, tumor grade or malignancy grading score. We conclude that methylation of the APC1A promoter in cervical cancer, as diagnosed by pyrosequencing, is significantly related to major biological characteristics of the tumor, and may be a new predictor of poor prognosis in cervical cancer.     

Clinical features and genetic analysis of Korean patients with Loeys-Dietz syndrome

Loeys-Dietz syndrome (LDS) is an inherited disorder that is characterized by the triad of arterial tortuosity and aneurysms, hypertelorism and a bifid uvula or cleft palate. The disease is caused by heterozygous mutations in the genes encoding transforming growth factor β receptors 1 and 2 (TGFBR1 and TGFBR2, respectively). However, studies of patients with LDS are limited in Korea. From June 2000 to December 2010, 13 patients (10 probands) diagnosed with LDS were enrolled. The multidisciplinary data of the patients were reviewed retrospectively. The frequency of each clinical manifestation in Korean patients with LDS was compared with Western populations as described in the report by Loeys et al. Twelve (92%) of the 13 LDS patients had arterial tortuosity, 9 (69%) patients had hypertelorism and 11 (85%) patients had bifid uvula or cleft palate. Mutations in either TGFBR1 or TGFBR2 were detected in nine probands (90%). Of the mutations, five novel mutations were detected; three in TGFBR2 and two in TGFBR1. Blue sclera and atrial septal defect were not observed in the Korean patients, and the frequency of blue sclera was significantly lower in our Korean population than previously-described Western population (0 vs 40%; P=0.005). Despite the restricted number of patients in our study, we identified five novel mutations in the TGFBR1 and TGFBR2 genes and, except for blue sclera, no differences in phenotype are apparent between Korean patients and Western patients.     

Detection of left atrial thrombus in patients with mitral stenosis and atrial fibrillation: retrospective comparison of two-phase computed tomography, transoesophageal echocardiography and surgical findings

Objective

This retrospective study aims to assess the accuracy of two-phase computed tomography (CT) and transoesophageal echocardiography (TEE) for the detection of left atrial (LA) thrombus in patients with mitral stenosis (MS) and atrial fibrillation (AF), by using intraoperative findings as the reference standard.

Methods

Preoperative two-phase CT and intraoperative TEE were performed in 106 patients with MS and AF. The ratio (LAA/AA_L) of Hounsfield units (HU) in the LA appendage (LAA) to the ascending aorta (AA) was calculated on the late-phase CT image.

Results

LA echodense masses on TEE and LA filling defects on two-phase CT were observed in 29 and 39 patients, respectively. Thirty-five LA thrombi were identified at surgery in 27 patients. Compared with the intraoperative findings, per-patient sensitivity, specificity, positive and negative predictive values of two-phase CT were 100 %, 85 %, 69 % and 100 %, and those by using TEE were 93 %, 95 %, 86 % and 97 % in detecting LAA thrombus. After adopting the cut-off value of 0.5 for the LAA/AA_L HU ratio, the specificity and positive predictive value of two-phase CT were increased to 96 % and 90 %, respectively.

Conclusion

Two-phase CT with a cut-off value of LAA/AA_L HU ratio of 0.5 provides high performance for the detection of LAA thrombus.

Key Points

?Accurate detection of left atrial appendage (LAA) thrombus is extremely important. ?However artefacts from flow effects influence both CT and ultrasound findings. ?Two-phase ECG-gated CT offers new insight into thrombus detection. ?Analysis of aortic/atrial opacification helps differentiate LAA thrombus from artefact at CT.