Novel Dental Anomaly–associated Mutations in WNT10A Protein Binding Sites

ObjectiveWNT/β-catenin signaling is initiated by binding of a WNT protein to a Frizzled (FZD) receptor and a co-receptor, low-density lipoprotein (LDL) receptor-related protein 5 or 6 (LRP5/6). The objective of this study was to find the genetic variants responsible for dental anomalies found in 4 families.MethodsClinical and radiographic examination and whole exome sequencing were performed on 5 patients affected with dental anomalies and the mutant proteins modeled.ResultsFive patients were heterozygous for the WNT10A variants, including c.877C>T; p.Arg293Cys, c.874A>G; p.Ser292Gly, c.1042C>T; p.Arg348Cys, and c.1039G>T; p.347GluX. The p.Arg293Cys and p.Ser292Gly mutations are located in the WNT10A N-terminal domain region with binding sites for FZD receptor, porcupine, WNTLESS, and extracellular binding proteins, so they are likely to have adverse effects on binding these proteins. The p.Arg348Cys mutation, which is located in the binding site of LRP5/6 co-receptors, is postulated to result in impaired binding to these co-receptors. The nonsense mutation p.347GluX is predicted to result in the truncation of most of the C-terminal domain, which is likely to disrupt the binding of WNT10A to WNTLESS, the membrane protein that binds lipid-acylated WNT proteins to carry them from the endoplasmic reticulum to the cell surface and FZD.ConclusionsFour novel mutations in WNT10A were identified in patients with isolated tooth agenesis. The mutations in the N-terminal domain and the interface between the N- and C-terminal domains of WNT10A in our patients are likely to disrupt its binding with FZD, LRP5/6, and various other proteins involved in WNT10A processing and transport, impair WNT and SHH signaling, and subsequently result in tooth agenesis, microdontia, and root maldevelopment.Key words: WNT10A mutation, Frizzled binding site, Dental anomaly, Hypodontia, Dental malformation, Dental defect     

Optimal dose of nalbuphine for treatment of intrathecal-morphine induced pruritus after caesarean section.

OBJECTIVE: To search for the optimal dosage of nalbuphine relief of intrathecal-morphine induced pruritus after caesarean section. METHODS: Ninety parturients who developed moderate to severe pruritus caused by intrathecal morphine after caesarean section were randomly allocated into 3 groups receiving 2, 3 and 4 mg of intravenous nalbuphine respectively. The improvement of pruritus and adverse effects of nalbuphine such as increasing pain scores, nausea, vomiting, sedation and respiratory depression were evaluated at 15 minutes after nalbuphine administration. RESULTS: Percentage of successful treatment of pruritus with 2, 3 and 4 mg, nalbuphine were 86.7, 96.7 and 100, respectively (p = 0.12). There was no statistically significant difference in adverse effects. No evidence of respiratory depression was detected. However, there was significant increase in pain scores in group of 4 mg nalbuphine (p = 0.004). CONCLUSIONS: Nalbuphine of 2 to 3 mg was considered to be adequate in treatment of intrathecal morphine induced pruritus after caesarean section without increasing pain scores or causing other side effects.     

Tissue-Specific Characteristics of <Emphasis Type="Italic">in Vivo</Emphasis> Electric Gene: Transfer by Tissue and Intravenous Injection of Plasmid DNA

Purpose To evaluate the tissue-specific characteristics of electric gene transfer after tissue and intravenous injection of naked plasmid DNA (pDNA).Methods pDNA encoding firefly luciferase was injected directly into the liver, kidney, spleen, skin and muscle, or into the tail vein of mice, and electric pulses were then applied to one of these organs. The distribution of transgene expressing cells was evaluated using pDNA encoding -galactosidase.Results Tissue injection of pDNA produced a significant degree of transgene expression in any tissue with the greatest amount in the liver, followed by kidney and spleen. The expression in these organs decreased quickly with time, and muscle showed the greatest expression at 7 days. Electroporation significantly increased the expression, and the expression level was comparable among the organs. Intravenous injection of pDNA followed by electroporation resulted in a significant expression in the liver, spleen, and kidney but not in the skin or muscle.Conclusions Electric gene transfer to the liver, kidney, and spleen can be an effective approach to obtain significant amounts of transgene expression by either tissue or intravenous injection of pDNA, whereas it is only effective after tissue injection as far as skin- or muscle-targeted gene transfer is concerned.     

Molecular identification of Trichinella papuae from a Thai patient with imported trichinellosis

Previously, we reported the presence of imported trichinellosis in a Thai worker returning from Malaysia, who presented with progressive generalized muscle hypertrophy and weakness after eating wild boar meat. This work analyzed a partial small subunit of a mitochondrial ribosomal RNA gene of Trichinella larvae isolated from the patient. The results showed complete identity with a mitochondrial RNA gene of Trichinella papuae (GenBank accession no. EF517130). This is the first report of imported trichinellosis in Thailand caused by T. papuae. It is possible that T. papuae is widely distributed in the wildlife of Southeast Asia.     

Evaluating the in vitro efficacy of gatifloxacin,levofloxacin and gentamicin against Acanthamoeba cysts

International Ophthalmology - To evaluate the in vitro efficacy of three commercial ophthalmic solutions (gatifloxacin, levofloxacin and gentamicin) against cysts of Acanthamoeba species....     

Molecular identification of Ascaris lumbricoides and Ascaris suum recovered from humans and pigs in Thailand,Lao PDR,and Myanmar

Parasitology Research - Ascaris lumbricoides is the largest roundworm known from the human intestine while Ascaris suum is an internal parasite of pigs. Ascariasis, caused by Ascaris lumbricoides,...     

Nalbuphine versus ondansetron for prevention of intrathecal morphine-induced pruritus after cesarean delivery

In this prospective, randomized, double-blinded study, we compared the prophylactic efficacy of nalbuphine and ondansetron for the prevention of intrathecal morphine-induced pruritus after cesarean delivery. Two-hundred-forty parturients were randomly allocated into four groups. The N-4 group, O-4 group, O-8 group, and placebo group received IV 4 mg of nalbuphine, 4 mg of ondansetron, 8 mg of ondansetron, and 4 mL of normal saline, respectively, immediately after the baby was delivered. In the postanesthesia care unit, we found that the severity of pruritus score in the four groups was significantly different (P < 0.001). The prophylactic success rate for pruritus of the N-4, O-4, O-8, and placebo groups was 20%, 13%, 12%, and 6%, respectively (P < 0.001). The pruritus score between N-4 and placebo and O-4 and placebo was significantly different (P < 0.001 and P = 0.006, respectively). Treatment for pruritus was requested by patients in 25%, 47%, 51%, and 72% of patients in the N-4, O-4, O-8, and placebo groups, respectively (P < 0.001). There were no differences among groups in nausea/vomiting score, pain score, sedation score, or shivering score at 4, 8, and 24 h after surgery. Nalbuphine and ondansetron are more effective than placebo for the prevention of intrathecal morphine-induced pruritus after cesarean delivery. IMPLICATIONS: Nalbuphine and ondansetron are more effective than placebo for the prevention of intrathecal morphine-induced pruritus after cesarean delivery.     

Current Status of Human Hookworm Infections,Ascariasis, Trichuriasis,Schistosomiasis Mekongi and Other Trematodiases in Lao People's Democratic Republic

Soil-transmitted helminth (STH) infections, and schistosomiasis and other trematodiases often have a high prevalence in developing countries. Here, we present updated information on the prevalence of these parasites in the Lao People''s Democratic Republic (Lao PDR) in 2012, arising from the annual national helminthiasis survey. Fecal specimens were collected from 8,610 inhabitants of 12 provinces and one municipality (Bokeo, Houaphan, Luang Namtha, Luang Prabang, Oudomxay, and Phongsaly Provinces from northern Lao PDR; Bolikhamxay and Xieng Khouang Provinces and Vientiane Municipality from the central part of the country; and Attapeu, Champasak, Saravan, and Sekong Provinces from southern Lao PDR). The overall prevalence of three major STHs, Ascaris lumbricoides, Trichuris trichiura, and hookworms (Necator americanus and Ancylostoma duodenale) were 11.6%, 8.5%, and 25.0%, respectively. Prevalence of Schistosoma mekongi infection was 0.1%, and of miscellaneous trematodiases (including opisthorchiasis) was 14.0%. Clearly, the nationwide parasite control project is still necessary to reduce morbidity caused by helminthic diseases.Three major soil-transmitted helminths (STHs), i.e., roundworms (Ascaris lumbricoides), whipworms (Trichuris trichiura), and hookworms (Necator americanus and Ancylostoma duodenale) are important public health problems, mostly impacting the health of children in several countries of the Americas, sub-Saharan Africa, China, and East and Southeast Asia, including Lao People''s Democratic Republic (Lao PDR).^1–3 Ascaris lumbricoides is estimated to infect 1.221 billion people globally, T. trichiura 795 million, and hookworms, 740 million globally.¹ The blood fluke, Schistosoma mekongi, occurs in communities along the Mekong River in Cambodia and Lao PDR,⁴ causing intestinal and hepatosplenic disease.^5,6 Many other trematode species can also infect people in Lao PDR. These include the highly pathogenic liver fluke, Opisthorchis viverrini, heterophyids such as Haplorchis taichui, and lecithodendriids such as Phaneropsolus bonnei, and Prosthodendrium molenkampi.⁷ To help planning for improvement of control strategies and evaluation of existing control activities, there was a need to update information on the prevalence and distributions of STHs and schistosomiasis and other trematodes. The information is presented here.During parasitological surveys in 2012, fecal specimens were collected from 8,610 inhabitants located in 12 provinces and one municipality (Bokeo [N = 537], Houaphan [N = 780], Luang Namtha [N = 440], Luang Prabang [N = 157], Oudomxay N = 969], and Phongsaly Provinces [N = 688] from northern Lao PDR; Bolikhamxay [N = 611]and Xieng Khouang [N = 1,143] Provinces and Vientiane Municipality [N = 243] from the central part of the country and Attapeu [N = 645], Champasak [N = 1,076], Saravan [N = 624], and Sekong Provinces [N = 697] from southern Lao PDR) (Figure 1). Sample size calculation was performed by using simple random sampling based on estimated preliminary prevalence data for STHs (Laymanivong and others, unpublished data). A total of 58 villages from 29 districts were randomly selected (2 or 3 districts per province and 2–3 villages per district). Approximately 300 inhabitants, with equal sex ratio, were randomly selected per village. The study was approved by the Ministry of Public Health, Lao PDR. Oral informed consent was obtained from all human adult participants and from parents or legal guardians of minors.Open in a separate windowFigure 1.Percentage prevalence of Ascaris lumbricoides (As), Trichuris trichiura (Tt), hookworm (Hw), Schistosoma mekongi (Sm), and miscellaneous trematodes (including Opisthorchis viverrini) (Mt) in Lao People''s Democratic Republic (PDR) in year 2012.Fecal specimens were examined twice for helminth eggs using the Kato-Katz thick smear technique. Parasitic infections were recorded; numbers of eggs for each helminth species were counted, and intensity of infection was calculated and expressed as eggs per gram of feces (epg). The overall prevalence of A. lumbricoides, T. trichiura, and hookworms were 11.6%, 8.5%, and 25.0%, respectively. The proportion of individuals infected with S. mekongi was 0.1%. Because it is difficult to differentiate eggs of the other trematode species from each other using the Kato-Katz thick smear technique,⁷ such eggs were recorded as miscellaneous trematode eggs. The overall prevalence of flukes producing such eggs was 14.0%. The individual prevalence of parasites in 12 provinces and one municipality are shown in Figure 1. Infections were categorized as light, moderate, or heavy, following previous criteria⁸ and are presented in Supplemental Table. The majority of infections were light. Before the implementation of a 5-year project⁹ supported by Loa PDR-Korea and the World Health Organization (WHO) Regional Office for the Western Pacific on the control of intestinal parasitic diseases for primary school in Lao PDR was commenced in 2000, the baseline data on the high prevalence of intestinal parasitic infections in Lao PDR were 34.9% for A. lumbricoides, 19.1% for hookworm, and 25.8% for T. trichiura.⁹ This study revealed that the prevalence of A. lumbricoides and T. trichiura infections decreased about 70% and parasite loads are also low. This reason could be a result of the good effective approach of a nationwide parasite control project.⁹

Table 1

Prevalences and intensities of Ascaris lumbricoides, Trichuris trichiura, hook worm, Schistosoma mekongi, and miscellaneous trematode infections in LAO PDR in year 2012