Human papillomavirus genotypes among infected Thai women with different cytological findings by analysis of E1 genes

Cervical cytological data may not be sufficient for cervical cancer screening and prevention. In this project, we determined HPV genotype among infected Thai women with different cytological findings by characterization of E1 genes. Five hundred and thirty-five specimens were tested by PCR amplification of the E1 genes. HPV genotypes were determined by sequencing, comparison with the GenBank database and were analyzed in relation to different cytological findings. HPV-DNA by PCR were typed and revealed 32 different genotypes. HR-HPV (HPV16, 18 or 52) was detected in all samples with cervical cancer cytology. HPV16 was most prevalent irrespective of cervical cytology. Moreover, HPV31 and 52 were most prevalent in the HSIL and LSIL groups whereas HPV66 was found mostly in the LSIL group. The LSIL group displayed the highest variation of HPV genotypes. Moreover, HPV31 and 52 predominated in the HSIL and LSIL groups especially HPV52 which was found in cancer samples. We hoped that these data of HPV genotypes can be used as preliminary data of HPV in Thailand and can serve as basic data for future research into the HPV genotype in south-east Asia.     

Polyherbal formulation exerts wound healing,anti-inflammatory,angiogenic and antimicrobial properties: Potential role in the treatment of diabetic foot ulcers

Diabetic foot ulcer (DFU) is a common and devastating complication in diabetic patients and is associated with an elevated risk of amputation and mortality. DFU remains a major therapeutic challenge due to poor understanding of its underlying pathogenesis. This complication is characterized by impaired wound healing; however, mechanisms causing this impairment are complicated and involve interactions between many different cell types and infections. In addition to other conventional DFU treatments, herbal foot baths are also common, although little is known about their mechanisms of action, and they contain a wide variety of herbal ingredients. In this study, we aimed to examine the effects of three polyherbal formulations consisting of medicinal plants used in traditional Thai herbal foot baths on wound healing, anti-inflammation, angiogenesis, and extracellular matrix modulation using high-concentration glucose-treated human keratinocytes, in addition to antibacterial evaluation. Our results showed that formulation 3 (F3) possessed the greatest potential to restore the impairment of keratinocytes caused by high glucose concentrations. We found that F3 could inhibit the growth of Staphylococcus aureus, accelerate wound healing, and upregulate the expression of TIMP-1, VEGF, and TGF-β, and downregulate the expression of TNF-α, IL-6, and MMP-9. Collectively, these data support the potential of F3 for therapeutic development in the treatment of DFU.     

Proteomic profiling of cholangiocarcinoma cell line treated with pomiferin from Derris malaccensis

Pomiferin, a prenylated isoflavonoid from Derris malaccensis with strong anti-fungal and anti-oxidant activities, showed cytotoxic activity towards human cholangiocarcinoma cells (HuCCA-1), with IC(50) of 0.9 microg/mL. Pomiferin caused apoptosis, detectable by DNA fragmentation. Two-dimensional PAGE showed increased expression of 12 proteins, namely glucose-regulated protein 75 (grp 75), calcyclin (S100A6), degraded cytokeratin 19, ATP synthase D, ribosomal protein P0, degraded cytokeratin 18 (two spots pI/MW 6.03/29.9 and pI/MW 4.66/21.5), cofilin, annexin A1, triose phosphate isomerase, peroxiredoxin-1, calgizzarin, and profilin. In contrast, cytokeratins (CK) 7, 18 and 19 were down-regulated, and were shown by 1-DE immunodetection to be degraded.     

Renal tubular cell membranes inhibit growth but promote aggregation of calcium oxalate monohydrate crystals

Cell membranes have been proposed to serve as promoters for calcium oxalate monohydrate (COM) kidney stone formation. However, direct evidence to demonstrate the modulatory effects of renal tubular cell membranes on COM crystals does not currently exist. We thus examined the effects of intact MDCK cells and their fragmented membranes on COM crystal growth, aggregation and transformation. COM crystals were generated in the absence (control) or presence of intact MDCK cells or their membrane fragments. Intact MDCK cells and their membrane fragments significantly inhibited COM crystal growth (22.6% and 25.2% decreases in size, respectively) and significantly reduced COM total crystal mass (23.1% and 25.6% decreases, respectively). In contrast, both of them markedly promoted crystal aggregation (1.9-fold and 3.2-fold increases, respectively). Moreover, both intact cells and membrane fragments could transform COM to calcium oxalate dihydrate (COD) crystals. Finally, COM crystal growth inhibitory activities of both membrane forms were successfully confirmed by a spectrophotometric oxalate-depletion assay. Our data provide the first direct evidence to demonstrate the dual modulatory effects of MDCK membranes on COM crystals. Although growth of individual COM crystals was inhibited, their aggregation was promoted. These findings provide additional insights into the mechanisms of COM kidney stone formation.     

Swine Is a Possible Source of Hepatitis E Virus Infection by Comparative Study of Hepatitis A and E Seroprevalence in Thailand

Hepatitis A virus (HAV) and hepatitis E virus (HEV) infection in developing countries are associated with contaminated food or water. Although Thailand is non-endemic for HEV, sporadic infections may occur from zoonotic transmission. Individuals between 7 months to 69 years (mean age = 32.8) from predominantly Islamic Narathiwat (n = 305) and swine farm-dense Lop Buri (n = 416) provinces were screened for anti-HEV and anti-HAV antibodies by commercial enzyme-linked immunosorbent assay and automated chemiluminescent microparticle immunoassay, respectively. Seroprevalence and relative antibody titers were analyzed according to age groups. HAV IgG antibody positive rates in Lop Buri and Narathiwat residents were 39.9% and 58%, respectively (p < 0.001). Greater than 90% of individuals >50 years old in both provinces possessed anti-HAV IgG. In contrast, seroprevalence for anti-HEV IgG was much higher in Lop Buri (37.3%) than in Narathiwat (8.9%) (p < 0.001). Highest anti-HEV IgG prevalence was found among 21-30 year-olds (50%) in Lop Buri and 41-50 year-olds (14.1%) in Narathiwat. In summary, fewer individuals possessed anti-HEV IgG in Narathiwat where most residents abstained from pork and fewer swine farms are present. Therefore, an increased anti-HEV IgG seroprevalence was associated with the density of swine farm and possibly pork consumption. Adults were more likely than children to have antibodies to both HEV and HAV.     

Decolorization of Molasses Waste Water by a Thermophilic Strain,Aspergillus fumigatus G-2-6 Sadahiro

We screened for fungi that can decolorize molasses melanoidin in the tropical zone and isolated some strains, mainly in the genus Aspergillus. Of these, strain No. G-2–6 was most active and was identical with Aspergillus fumigatus based on detailed morphological studies.

This strain decolorized about 75% of a molasses melanoidin solution when the strain was cultivated on a glycerol-peptone medium at 45°C for 3 days with shaking. In successive decolorization reusing the mycelia, this strain had more than 60% of the melanoidin-decolorizing activity at the eighth replacement in the presence of 4% glycerol.

Continuous decolorization of molasses melanoidin solution in a jar fermentor had an almost constant decolorization yield of about 70% at a dilution rate of 0.014 hr^-1. At the same time, about 51 % of the chemical oxygen demand and 56% of the total organic carbon in the initial solution were removed. In contrast, continuous decolorization of non-dialyzed molasses melanoidin solution removed a little more chemical oxygen demand and total organic carbon than those of dialyzed molasses melanoidin solution, but had a lower level of melanoidin-decolorizing activity (about 40%).     

Bacteria can promote calcium oxalate crystal growth and aggregation

Our previous report showed that uropathogenic bacteria, e.g., Escherichia coli, are commonly found inside the nidus of calcium oxalate (CaOx) kidney stones and may play pivotal roles in stone genesis. The present study aimed to prove this new hypothesis by direct examining CaOx lithogenic activities of both Gram-negative and Gram-positive bacteria. CaOx was crystallized in the absence (blank control) or presence of 10⁵ CFU/ml E. coli, Klebsiella pneumoniae, Staphylococcus aureus, or Streptococcus pneumoniae. Fragmented red blood cell membranes and intact red blood cells were used as positive and negative controls, respectively. The crystal area and the number of aggregates were measured to initially screen for effects of bacteria on CaOx crystal growth and aggregation. The data revealed that all the bacteria tested dramatically increased the crystal area and number of crystal aggregates. Validation assays (spectrophotometric oxalate-depletion assay and an aggregation–sedimentation study) confirmed their promoting effects on both growth (20.17 ± 3.42, 17.55 ± 2.27, 16.37 ± 1.38, and 21.87 ± 0.85 % increase, respectively) and aggregation (57.45 ± 2.08, 51.06 ± 5.51, 55.32 ± 2.08, and 46.81 ± 3.61 % increase, respectively) of CaOx crystals. Also, these bacteria significantly enlarged CaOx aggregates, with the diameter greater than the luminal size of distal tubules, implying that tubular occlusion might occur. Moreover, these bacterial effects were dose-dependent and specific to intact viable bacteria, not intact dead or fragmented bacteria. In summary, intact viable E. coli, K. pneumoniae, S. aureus, and S. pneumoniae had significant promoting effects on CaOx crystal growth and aggregation. This functional evidence supported the hypothesis that various types of bacteria can induce or aggravate metabolic stone disease, particularly the CaOx type.     

Density functional investigation of hydrogen gas adsorption on Fe?doped pristine and Stone?Wales defected single?walled carbon nanotubes

The adsorptions of hydrogen molecule of the Fe?-?doped pristine and Stone?-?Wales defected armchair (5,5) single?-?walled carbon nanotubes (SWCNTs) compared with the pristine SWCNT were investigated by using the density functional theory at the B3LYP/LanL2DZ level. The doping of Fe atom into SWCNTs occurring via an exothermic process was found. The adsorptions of hydrogen molecule on the Fe?-?doped structures of either perfect or SW defected SWCNTs are stronger than on their corresponding undoped structures. The structural and electronic properties of the pristine and SW defected SWCNTs, their Fe?-?doped structures and their hydrogen molecule adsorptions are reported.