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Development of a point-of-care test to detect hepatitis B virus DNA threshold relevant for treatment indication

Abstract

Background

Hepatitis B virus (HBV) has been the most prevalent blood-borne pathogen wherein utero transmission has still not been properly managed. Recent practice guidelines suggested that an antiviral drug should be administered to third-trimester pregnancies with significant viremia (>2 × 105 IU/mL).

Objectives

To develop a novel turbidity-based loop-mediated isothermal amplification (LAMP) coupled with heat treatment DNA extraction method that is a rapid, cost-effective, and feasible viral load assessment and could be applied to antenatal screening.

Methods

Primers and reagents were designed, turbidity-based platform and heat treatment method were added, and evaluated for optimal efficiency. Assay sensitivity was tested from serially diluted standard HBV DNA. Assay specificity was tested with six standard viral DNAs. Clinical samples were analyzed and the results were compared with those of quantitative polymerase chain reaction (qPCR) diagnostic records.

Results

The optimized condition was 60°C with no betaine, 1.4 mM deoxyribonucleotide triphosphates (dNTPs) and 6 mM of MgSO4 for 60 min. The assay accurately detected samples with standard HBV DNA at >2 × 105 IU/mL in both distilled water and spiked serum. Results can be interpreted within 31.48 ± 1.41 min in real-time turbidimeter. The amplification is exclusively specific to HBV, but not with the other six human-specific viruses. Moreover, the assay showed comparable performance within 95% confidence interval to the previously developed HBV LAMP toward clinical specimens.

Conclusions

This newly developed method was accurate, affordable, and flexible to further implementation to large-scale third-trimester pregnancy screening.

Open access
Evidence of growth hormone effect on plasma leptin in diet-induced obesity and diet-resistant rats

Abstract

Background

Plasma leptin is regulated by several factors, including growth hormone (GH), which influences the pathophysiology of obesity.

Objective

To demonstrate the short-term effect of GH on plasma leptin levels in 3 conditions in vivo with the different amount of body fat mass.

Methods

Adult male Wistar rats were fed with standard chow or hypercaloric diet (HC). The HC rats were demonstrated as HC-feeding obese (HC-O) and HC-feeding resistant (HC-R) rats. Then, they were treated with GH or saline for 3 days. Basal plasma leptin levels were measured at 24 and 32 h. For meal-induced condition, all rats were fed for 2 hand plasma leptin was measured. Further 16-h fasting period, plasma leptin, insulin, and insulin sensitivity indexes were determined.

Results

The short-term GH treatment decreased basal plasma leptin at 32 h after the first GH injection in HC-O rats. However, GH treatment had no effect on meal-induced plasma leptin in all rats. Furthermore, GH treatment attenuated fasting effect on plasma leptin in control and HC-R rats. The insulin resistance (IR) induced by the short-term GH treatment was demonstrated by higher fasting plasma insulin and the increased homeostasis model of IR in HC-R rats.

Conclusions

The study demonstrates the important role of greater fat mass in HC-O rats, which results in decreased basal plasma leptin after short-term GH treatment. For meal-induced condition, GH had no effect on plasma leptin in all rats. Interestingly, GH could attenuate fasting effect on plasma leptin in rats that have lower fat mass.

Open access
Lactobacillus plantarum B7 attenuates Salmonella typhimurium infection in mice: preclinical study in vitro and in vivo

Abstract

Background

Salmonella typhimurium is a cause of gastroenteritis including diarrhea. Lactobacillus plantarum is a probiotic widely used to prevent and treat diarrhea.

Objectives

To determine the protective effects of L. plantarum B7 on diarrhea in mice induced by S. typhimurium.

Methods

Inhibition of S. typhimurium growth by L. plantarum B7 was determined using an agar spot method. Mice were divided into 3 groups (n = 8 each): a control group, an S group administered 3 × 109 CFU/mL S. typhimurium, and an S + LP group administered 1 × 109 CFU/mL L. plantarum B7 and 3 × 109 CFU/mL S. typhimurium daily for 3 days. Counts of S. typhimurium and percentage of fecal moisture content (%FMC) were determined from stool samples. Serum levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and CXCL1 were determined.

Results

L. plantarum B7 produced a clear zone on S. typhimurium. There were significantly less S. typhimurium in the feces from mice in the S+LP group than in the S group. Serum levels of TNF-α, IL-6, and CXCL1 in mice from the S group were significantly higher than levels in the S+LP and control groups. Feces from mice in the S group were soft and loose, whereas in the S+LP group they were hard and rod shaped. The %FMC in the S+LP group was significantly less than in the S group.

Conclusions

L. plantarum B7 can inhibit growth of S. typhimurium, decrease levels of proinflammatory cytokines, and attenuate symptoms of diarrhea induced in mice by S. typhimurium.

Open access
Outcomes of pancreaticoduodenectomy in patients with obstructive jaundice with and without preoperative biliary drainage: a retrospective observational study

Abstract

Background

Preoperative biliary drainage (PBD) in patients with obstructive jaundice from periampullary neoplasms may reduce the untoward effects of biliary obstruction and subsequent postoperative complications. However, PBD is associated with bile contamination and increases infectious complications after pancreaticoduodenectomy (PD).

Objectives

To determine whether PBD is associated with more complications after PD.

Methods

Patients with obstructive jaundice from periampullary lesions who underwent PD from 2000 to 2015 at our institution were retrospectively enrolled. The cohort was divided into a group with PBD and a group without. PBD was performed using one of the following methods: endoprosthesis, percutaneous transhepatic biliary drainage, surgical biliary-enteric bypass, or T-tube choledochostomy. PDs were performed by the first author using uniform surgical techniques. Postoperative complications were recorded. Statistical analyses were conducted using an unpaired t, Fisher exact, or chi-squared tests as appropriate.

Results

There were 26 with PBD and 28 patients without. Patients in the 2 groups were similar in age, presenting serum bilirubin level, operative time, operative blood transfusion, and hospital stay. The group with PBD had longer duration of jaundice, more patients presenting with cholangitis, and more patients with carcinoma of the ampulla of Vater. The overall complications were higher in patients in the group with PBD than in the group without.

Conclusions

PBD was associated with more complications overall after PD. However, PBD was necessary and lifesaving in certain clinical situations and improved the condition of patients before they underwent PD. Routine PBD in patients with obstructive jaundice without definite indications is not recommended.

Open access
Open access
Sodium glycerophosphate and prescribed calcium concentrations in pediatric parenteral nutrition: a retrospective observational study and economic evaluation

Abstract

Background

The risk of precipitation limits calcium and phosphate concentrations that can be administered parenterally to pediatric patients. As an alternative to dipotassium phosphate, sodium glycerophosphate (NaGlyP) is claimed to reduce the risk of precipitation in solutions for parenteral administration.

Objectives

To determine the calcium concentrations, NaGlyP, and dipotassium phosphate prescribed in pediatric parenteral nutrition orders and the cost–benefit of the organic phosphate.

Methods

We retrospectively collected cross-sectional data for parenteral nutrition orders from September 2014 to August 2015 for pediatric patients including neonates and children aged <18 years who were admitted to King Chulalongkorn Memorial Hospital, Bangkok, Thailand. Calcium concentration, calcium concentration adjustments, and costs of phosphate used per bag were analyzed.

Results

Of 2,192 parenteral nutrition orders, NaGlyP was used in 2,128 (97.1%) with calcium concentrations in the range of 0.84–139.91 mmol/L, which were significantly higher than calcium concentrations used with dipotassium phosphate (0.00–12.21 mmol/L, P < 0.001). There was no report of visible precipitation. Median costs of NaGlyP and dipotassium phosphate used per unit bag were not significantly different (35.88 and 41.25 Thai baht [THB] or 1.04 and 1.20 USD per bag, respectively, P>0.99; (1 USD equivalent to 34.241 THB U.S. Federal Reserve Bank G5.A annual average rate 2015).

Conclusions

Higher calcium concentrations could be achieved without increasing the direct cost per unit bag significantly as a result of using NaGlyP, an alternative to dipotassium phosphate as a source of phosphate for patients who require high amounts of calcium in parenteral nutrition.

Open access
21st Century era of anatomy
Open access
Anatomy of the vasculature supplying hepatobiliary structures and celiac trunk branching patterns in the Thai population

Abstract

Background

Knowledge of the anatomy of the celiac trunk (CT) and arterial supply of the hepatobiliary system is essential for surgical and interventional radiological treatment of upper abdominal diseases.

Objectives

To determine the branching patterns of the CT and variation in origin and type of the right hepatic artery (RHA), left hepatic artery (LHA), and cystic artery (CA).

Methods

The anatomy of the CT in 100 cadavers from Thai adult donors was observed in 3 aspects: its branching pattern, the origin of the RHA and LHA, and the origin of the CA and its relation to the common bile duct (CBD) and common hepatic duct (CHD).

Results

The majority of the CT branching pattern was categorized as the type II classical pattern, which has 3 branches: the left gastric artery (LGA), splenic artery (SA), and common hepatic artery (CHA). The RHA branched from proper hepatic artery in 67 cadavers. The origin of the accessory RHA was either from the abdominal aorta or superior mesenteric artery (SMA), whereas the replaced RHA originated from the CHA, SMA, or CT. The accessory LHA ramified from CHA (2 cases) and LGA (1 case). The replaced LHA was found in 30 cadavers and 29 arose from the CHA. The single and double types of CA were found in 94 and 4 cadavers, respectively. In all, 57% of single CA passed posteriorly and 39% passed anteriorly to the CBD and CHD.

Conclusions

To lower posttreatment complications, variations in the anatomy and the vascular supply of hepatobiliary structures should be considered.

Open access
Behavioral and histopathological studies of cervical spinal cord contusion injury in rats caused by an adapted weight-drop device

Abstract

Background

Models of spinal cord injury (SCI) caused by weight-drop devices to cause contusion have been used extensively, and transient behavioral deficits after thoracic injury have been demonstrated. The severity of the injury caused by the device should be mild enough to allow recovery.

Objective

To determine whether our adapted weight-drop device with a small tip can effectively induce mild hemicontusion at the level of the fifth cervical vertebra.

Methods

We divided 15 adult male Sprague Dawley rats into groups of 5 for the following treatments: sham (SH, laminectomy only), mild (MSCI) or severe SCI (SSCI). Behavioral tests and histopathology were used before (day 1) and after the treatment on days 3, 7, 14, 21, 28, and 35 to assess the injury.

Results

Rats with SSCI showed a significant somatosensory deficit on days 3 and 7 compared with rats in the SH group, recovering by day 14. In a horizontal-ladder test of skilled locomotion, rats with SSCI showed a significant increase in error scores and percentage of total rungs used, and a decrease in the percentage of correct paw placement compared with rats in the SH group. There was greater recovery to normal paw placement by rats with MSCI than by rats with SSCI. These behavioral deficits were consistent with histopathology using hematoxylin and eosin counterstained Luxol fast blue, indicating the degree of injury and lesion area.

Conclusions

Mild hemicontusion caused by the adapted device can be used to evaluate SCI and provides a model with which to test the efficacy of translational therapies for SCI.

Open access
Changes in sperm quality and testicular structure in a rat model of type 1 diabetes

Abstract

Background

Chronic hyperglycemia is a characteristic of diabetes mellitus (DM). Long-lasting hyperglycemia can generate oxidative stress and reactive oxygen species. The effect of this condition on sperm quality and spermatogenesis leads to male infertility and reproductive dysfunction.

Objectives

To investigate changes in sperm quality, morphology of testicular structure, and stage of development of seminiferous tubules in a streptozotocin (STZ)-induced rat model of type 1 DM.

Methods

We divided 15 male Sprague Dawley rats into 2 groups. DM was induced in 7 rats using STZ (60 mg/kg intraperitoneally), while the other 8 were treated with citrate buffer as a vehicle control group. Rat semen was collected for quality measurements including motility, normal morphology, and concentration. Morphological changes in testicular structure and stage of development of seminiferous tubules were investigated by histology with hematoxylin and eosin (HE) staining.

Results

Significant decreases in all parameters of sperm quality and testicular weight were found in rats with induced DM. Moreover, abnormal morphology of seminiferous tubules including separation of the germinal epithelium, vacuolization, luminal sloughing of germ cells, and tubular atrophy was increased significantly in these rats, while the proportion of their seminiferous tubules at an early stage of development was significantly higher, but was dramatically decreased in the late stage of development when compared with that in vehicle-treated control rats.

Conclusions

DM has adverse effects on sperm quality, testicular structure, and development of seminiferous tubules. These findings may reflect the male infertility and reproductive dysfunction seen in patients with type 1 DM.

Open access