Rapid detection of the most common high-risk human papillomaviruses by loop-mediated isothermal amplification

Persistent infection with high-risk human papillomavirus (HPV) is a major risk factor for development of cervical cancer. At present, polymerase chain reaction (PCR)-based methods, the most widely molecular tools used for HPV detection, are time-consuming and require expensive instruments. In this study, loop-mediated isothermal amplification (LAMP) was established for detection of HPV types 16, 18, 45 and 58 which are frequently found in Thailand. The optimal condition for detection of these high risk HPVs was 63°C for 60min. Since a white magnesium pyrophosphate precipitate is a characteristic by product of the LAMP reaction which can be visualized directly by the naked eye, the entire assay time of LAMP is 1h compared to 6-8h of for a nested PCR detection. The detection limit of LAMP assay was shown to be equivalent to nested PCR that could amplify 10(2) copies of HPV-18 and 10(3) copies of HPV 16, 45 and 58, as determined by either turbidity detection or agarose gel electrophoresis. No cross-reaction was observed, indicating that LAMP assay has high type-specificity. The assay showed successful detection of HPV in 56 clinical specimens. Using nested PCR as the gold standard, the sensitivity, specificity, negative predictive values and positive predictive values of LAMP assay were 100%. In conclusion, LAMP assay is a high efficiency, low cost diagnostic tool, useful for rapid, accurate, direct detection of HPV for clinical diagnosis.     

Renal abscess in healthy children: report of three cases

We report three cases of renal abscess in otherwise healthy boys. One had a history of blunt trauma and two of antecedent skin infection. All presented with flank pain and fever. Urine and blood cultures were sterile. The diagnosis was made using ultrasound and computerized axial tomography. The first patient had recurrent renal abscess, following surgery and a short course of antibiotics. Eventually all responded well to long-term antibiotics only.     

Utility of QT interval corrected by Rautaharju method to predict drug-induced torsade de pointes

Introduction: New QT correction formulae derived from large populations are available such as Rautaharju’s [QTcRTH?=?QT * (120?+?HR)/180] and Dmitrienko’s [QTcDMT?=?QT/RR^0.413]. These formulae were derived from 57,595 and 13,039 cases, respectively. Recently, a study has shown that they did not experience errors across a wide range of heart rates compared to others.

Objectives: (1) To determine the best cut-off value of QTcRTH and QTcDMT as a predictor of torsade de pointes (TdP) and (2) to compare the sensitivity and specificity using the cut-off value of QTcRTH with those of the QTcBazett (QTcBZT), QTcFridericia (QTcFRD), and QT nomogram.

Methods: Data were derived from two data sets. All cases aged over 18 years with an exposure to QT-prolonging drugs. Group-1, all cases developed TdP. Data in Group-1 were obtained from systematic review of reported cases from Medline since its establishment until 10 December 2015. Group-2 is composed of those who overdosed on QT prolonging drugs but did not develop TdP. This data set was previously extracted from a chart review of three medical centers from January 2008 to December 2010. Data from both groups were used to calculate QTcRTH and QTcDMT. The cut-off values from QTcRTH and QTcDMT that provided the best sensitivity and specificity to predict TdP were then selected. The same method was applied to find those values from QTcBZT, QTcFRD, and QT nomogram. The receiver operating characteristic curve (ROC) was applied where appropriate.

Results: Group-1, 230 cases of drug-induced TdP were included from the systematic review of Medline. Group-2 (control group), which did not develop TdP, consisted of 292 cases. After applying all of the correction methods to the two datasets, the best cut-off values that provided the best accuracy (Ac) with the best sensitivity (Sn) and specificity (Sp) for each formula were as follows: QTcRTH at 477 milliseconds (ms), Ac?=?89.08%, Sn?=?91.30% (95%CI?=?86.89–94.61), Sp?=?87.33%(95%CI?=?82.96–90.92); QTcDMT at 475?ms, Ac?=?88.31%, Sn?=91.30% (95%CI?=?86.89–94.61), Sp?=?85.96%(95%CI?=?81.44–89.73); QTcBZT at 490?ms, Ac?=?86.97%, Sn?=?88.26% (95%CI?=?83.38–92.12), Sp?=?85.96% (95%CI?=?81.44–89.73); QTcFRD at 473?ms, Ac?=?88.89%, Sn?=?89.13% (95%CI?=?84.37–92.84), Sp =88.70% (95%CI?=?84.50–92.09). We found a significant difference (p-value?=?0.0020) between area under the ROC of the QTcRTH (0.9433) and QTcBZT (0.9225) but not QTcFRD (0.9338). The Ac, Sn, and Sp of the QT nomogram were 89.08%, 91.30% (95%CI?=?86.89–94.61), and 87.33% (95%CI?=?82.96–90.92), respectively, and they were all equal to those of QTcRTH.

Conclusion: Rautaharju method not only produced minimal errors for QT interval correction but also at QTcRTH 477?ms, it could predict TdP as accurately as QT nomogram and was better than the QTcBZT.     

Laser tattoo removal in Thai students

Objective

To review the results of laser tattoo removal in Thai students.

Material and methods

This is a retrospective review study of the students participating in our tattoo removal program at Srakaew Province, Thailand. The laser used was a 1064 nm Q-switched Nd:YAG laser (spot size: 3 mm, pulse duration: 6 ns, fluence: 3.5–7.5 J/cm²). A maximum of 5 laser treatments per student was performed at 2-month intervals.The data collected included age and sex of the students and information regarding the tattoos (time of tattoo existence, person who performed the tattoo procedure, method and price of tattooing, number of tattoo figures, complications during and after tattoo procedure including cleaning methods of tattoo needle) and the laser procedure (laser parameters, pain level during laser removal assessed by pain score sheet, treatment results, complications, side effects). Correlations were analyzed between the person who did the tattoo and complications after laser tattoo removal procedure, the age of tattoo and results of laser treatment and complications, and between the pain level and the time of EMLA^® application.

Results

The 115 students involved were aged between 8 years 4 months and 19 years 11 months. The mean time of tattoo existence was 1.9 ± 1.2 years. The majority of the students were tattooed (1) by amateur practitioners and (2) by hand. The costs of tattoo procedure ranged between 0 and 118 Thai Bahts. Friends influenced the students in favor of tattooing, and both the home and school were popular places for the tattooing procedure to take place. Ankles and fingers were the most common sites for students’ tattoos. A clinical clearance of 76–100% was achieved in 75% of the treated tattoos. Observed side effects were mild to moderate. Textural changes, hyperpigmentation, hypopigmentation, and scarring occurred in 41%, 3%, 3% and 17% of the cases, respectively. There was no statistically significant correlation between EMLA^® application time and the pain score. There was also no correlation between the person who did the tattoo, or the age of tattoo to the complications or side effects after laser tattoo removal.

Conclusion

The students were influenced and had tattoos by persons familiar to them, mostly at places which they often attended, including temples and schools. The students were not primarily concerned about the social consequence of having tattoos. Laser tattoo removal by means of a Q-switched Nd:YAG laser could clinically clear tattoos in 75% of the cases. However, there were side effects, such as hypopigmentation, hyperpigmentation and scarring.     

Alcohol and alkalosis enhance excystation of Opisthorchis viverrini metacercariae

The northeastern region of Thailand has long been known as an endemic area of the human liver fluke infection which is caused by Opisthorchis viverrini. Humans are infected by ingestion of uncooked cyprinoid fish in traditional dishes such as “koi-pla,” “pla-som,” “pla-jom,” and “pla-ra.” To date, the prevalence of this parasite infection remains high because of cultural behavior and local beliefs. The popular misunderstanding among people in this area is that alcohol, lemon juice, and fish sauce can kill the parasites. Thus, they believe that they can eat raw fish without the risk of infection. This study attempts to clarify the effects of ethyl alcohol and acidosis–alkalosis on O. viverrini metacercariae excystation. Metacercariae of O. viverrini were obtained from infected cyprinoid fish in a natural reservoir. Most metacercariae were obtained from small cyprinoid fish. Metacercariae were divided into three experimental groups and were treated with solutions containing four different concentrations of ethyl alcohol, four different concentrations of salt, and a range of acidic/basic pH. Metacercariae excystation was observed at the assigned times, and the data were then analyzed. Salt had no effect on excystation. Interestingly, the optimal conditions for O. viverrini excystation were pH 9 and 25 % ethyl alcohol. The present study suggests that raw fish should not be eaten while drinking alcohol or when consuming other ingredients with pH 9, because both alcohol and pH 9 could induce O. viverrini metacercariae excystation, leading to the early development of parasites in the hepatobiliary system.     

Congenital bronchoesophageal fistula and tracheoesophageal fistula with esophageal atresia.

A case of initial esophageal atresia and tracheoesophageal fistula in a female newborn, later complicated by pneumonia and a second bronchoesophageal fistula, is reported. She was treated surgically by closure of the tracheoesophageal fistula and by end-to-end esophago-esophageal anastomosis. An esophagram at 1 month of age was normal. Three months later she developed severe, persistent right lower lobe pneumonia that required intensive antibiotic therapy and respiratory support. Esophagography was repeated and revealed a second fistula between the right main-stem bronchus and the lower esophagus. The bronchoesophageal fistula was repaired, and a right lower lobectomy was performed. Postoperative recovery was uncomplicated. Histologic examination indicated that the fistula was congenital in origin. To the best of our knowledge, this is the first reported case of a congenital bronchoesophageal fistula coexisting with a tracheoesophageal fistula and esophageal atresia.     

Mekong malaria. II. Update of malaria, multi-drug resistance and economic development in the Mekong region of Southeast Asia

In an expansion of the first Mekong Malaria monograph published in 1999, this second monograph updates the malaria database in the countries comprising the Mekong region of Southeast Asia. The update adds another 3 years' information to cover cumulative data from the 6 Mekong countries (Cambodia, China/Yunnan, Lao PDR, Myanmar, Thailand, Viet Nam) for the six-year period 1999-2001. The objective is to generate a more comprehensive regional perspective in what is a global epicenter of drug resistant falciparum malaria, in order to improve malaria control on a regional basis in the context of social and economic change. The further application of geographical information systems (GIS) to the analysis has underscored the overall asymmetry of disease patterns in the region, with increased emphasis on population mobility in disease spread. Of great importance is the continuing expansion of resistance of P. falciparum to antimalarial drugs in common use and the increasing employment of differing drug combinations as a result. The variation in drug policy among the 6 countries still represents a major obstacle to the institution of region-wide restrictions on drug misuse. An important step forward has been the establishment of 36 sentinel sites throughout the 6 countries, with the objective of standardizing the drug monitoring process; while not all sentinel sites are fully operational yet, the initial implementation has already given encouraging results in relation to disease monitoring. Some decreases in malaria mortality have been recorded. The disease patterns delineated by GIS are particularly instructive when focused on inter-country distribution, which is where more local collaborative effort can be made to rationalize resource utilization and policy development. Placing disease data in the context of socio-economic trends within and between countries serves to further identify the needs and the potential for placing emphasis on resource rationalization on a regional basis. Despite the difficulties, the 6-year time frame represented in this monograph gives confidence that the now well established collaboration is becoming a major factor in improving malaria control on a regional basis and hopefully redressing to a substantial degree the key problem of spread of drug resistance regionally and eventually globally.