An overview of human helminthioses in Vietnam: Their prevention, control and lessons learnt.

In Vietnam, helminthioses remain a major threat to public health and contribute to the maintenance of poverty in highly endemic regions. Through increased awareness of the damaging effects caused by helminthioses, the Vietnamese government has implemented many national programs over the past 30 years for the prevention and control of the most important helminthioses, such as, lymphatic filariasis, soil transmitted helminths, food borne zoonotic helminths, and others. Various control strategies have been applied to reduce or eliminate these worms, e.g. mass drug administration, economic development, control of vectors or intermediate hosts, public health interventions through education, proper composting procedures for excreta potentially containing helminth eggs, and the expansion of food supply chains and improved technologies for the production and inspection of food products. These control measures have resulted in a significant reduction in the distribution and transmission of helminth infections and have improved the overall living conditions and health outcomes of the Vietnamese citizens. However, the persistence of several helminth diseases continues in some endemic areas, especially where poverty is widespread and local traditions include the consumption of raw foods, especially fish and meats. This manuscript provides an overview of the helminth infection prevention and control programs conducted in Vietnam, their achieved results, learned lessons, and future works.

Molecular and morphological characterization of Dollfustrema bagarii (Digenea: Bucephalidae) metacercariae from aquaculture channel catfish (Ictalurus punctatus) in northern Vietnam.

White grub metacercariae were found in the livers and kidneys of diseased specimens of an introduced channel catfish, Ictalurus punctatus (Rafinesque), in Vietnam. Based on morphological features and 28S rDNA sequence analysis, the isolated metacercariae were identified as Dollfustrema bagarii (Digenea: Bucephalidae) Moravec & Sey. Histopathological examination shows that encysted metacercariae can change the tissue structure of the infected organs and is often accompanied by haemorrhaging and the presence of eosinophilic granular cell infiltration. Degenerative changes were also observed.

Synthesis, resolution and crystal structures of two enanti-omeric rhodamine derivatives.

The title mol-ecule, rac-6'-bromo-3'-di-ethyl-amino-3H-spiro-[2-benzo-furan-1,9'-xanthen]-3-one, C24H20BrNO3, was synthesized and the two enanti-omers which formed were separated. The structures of all three compounds were determined and compared with those of a variety of related derivatives. A notable feature is the fold of the xanthene portion which ranges from 15.15 (13)° in the racemate to 2.42 (2)° in one mol-ecule of the R enanti-omer with that for the S enanti-omer having an inter-mediate value. The differences are attributed to the number and severity of inter-molecular inter-actions which include C-H⋯O hydrogen bonds, C-H⋯π(ring) and, in the S enanti-omer, a π-stacking inter-action between the carbonyl group and an aromatic ring.

The importance of wild fish in the epidemiology of Clonorchis sinensis in Vietnam.

Preliminary findings of a high prevalence of Clonorchis sinensis in wild-caught fish in a North Vietnam reservoir (Thac Ba reservoir, Yen Bai Province) prompted a longitudinal epidemiological study of fish infections. Monthly collections of fish from September 2014 to August 2015 were processed for recovery of metacercariae; 1219 fish, representing 22 species, were examined. Seven species were infected with C. sinensis metacercariae. Four species, Toxabramis houdemeri, Hemiculter leucisculus, Cultrichthys erythropterus, and Culter recurvirostris, had high prevalence (31.1 to 76.7 %); metacercarial intensities ranged from 3.9 to 65.7 metacercariae/fish. A seasonal variation of C. sinensis prevalence was observed in T. houdemeri. Variation in intensity of infection occurred in C. erythropterus and H. leucisculus. Intensity and prevalence of C. sinensis in the most highly infected species, T. houdemeri, varied by fish size; prevalence was higher in fish weighing more than 3 g, and intensity was higher in fish weighing more than 5 g. The distribution of metacercariae in the body region of T. houdemeri was significantly higher in the caudal fin (14.7 metacercariae/g), compared to the body and head regions (0.7 and 1.4 metacercariae/g, respectively). Further epidemiological investigations on C. sinensis in this reservoir region should include assessing the relative risk of the different fish species for humans based on the latter's food preferences, and the prevalence of C. sinensis in the community. The snail intermediate host(s) in the reservoir should also be identified along with the ecological factors influencing its exposure to C. sinensis eggs and its subsequent transmission of cercariae to fish. Also needed are investigations on the relative importance of wild and domestic reservoir hosts as sources of egg contamination of the reservoir.

Revision of Metahaliotrema Yamaguti, 1953 (Monogenoidea: Dactylogyridae), with new and previously described species from the spotted scat Scatophagus argus (Linnaeus) (Perciformes: Scatophagidae) in Vietnam.

An emended diagnosis of Metahaliotrema Yamaguti, 1953 (Monogenoidea: Dactylogyridae) is provided based on specimens of six species collected from the spotted scat Scatophagus argus (Linnaeus) (Scatophagidae) in Vietnam: M. scatophagi Yamaguti, 1953 (type-species); M. cf. yamagutii Mizelle & Price, 1964; M. mizellei Venkatanarasaiah, 1981; M. kulkarnii Venkatanarasaiah, 1981; M. ypsilocleithrum n. sp.; and M. similis n. sp. Methaliotrema filamentosum Venkatanarasaiah, 1981 from the whipfin silver-biddy Gerres filamentosus Cuvier (Gerreidae) is included as the only other valid member of the genus. Metahaliotrema arii Yamaguti, 1953 from an ariid catfish is considered incertae sedis within the Dactylogyridae; and Metahaliotrema srivastavai Singh & Agarwal, 1994 from a bagrid catfish is transferred to Chauhanellus Bychowsky & Nagibina, 1969 as Chauhanellus srivastavai (Singh & Agarwal, 1994) n. comb. Metahaliotrema geminatohamula Pan, Ding & Zhang, 1995 from spotted scat in China is determined to be a junior subjective synonym of M. scatophagi. The two new species and M. scatophagi, M. mizellei, and M. kulkarnii are described or redescribed based on specimens from Vietnam.

Two new axinid species (Monogenea: Axinidae) from the Pharao flyingfish Cypselurus naresii (Günther) (Beloniformes: Exocoetidae) in the Gulf of Tonkin off Vietnam.

A total of 21 Pharao flyingfish Cypselurus naresii (Günther) from the Gulf of Tonkin off Vietnam was examined for monogeneans. Ten individuals were parasitised by 72 specimens of two new axinid species of two rare and little known genera, Unnithanaxine Price, 1962 containing only one species, U. parawa (Unnithan, 1957), and Loxuroides Price, 1962 containing two species, L. sasikala (Unnithan, 1957) and L. fungilliformis Zhang, Ding, Liu & Wang, 1999. Unnithanaxine naresii n. sp. and Loxuroides pricei n. sp. are described and differentiated from the related species. Unnithanaxine naresii n. sp. is morphologically similar to U. parawa but is distinguished by the size of the clamps and reproduction organs, the number of spines in the lateral groups of the genital atrium, and in parasitism in a host fish species of a different genus. Loxuroides pricei n. sp. differs from L. fungilliformis in the greater size of the body, the number of clamps, testes, spines on cirrus and genital atrium, and in parasitism in a different host family. Similarly, L. pricei can be separated from L. sasikala in having a shorter distance from the anterior extremity to genital atrium or vaginal region, fewer testes, and a slightly greater number of spines on cirrus and genital atrium.

Enhanced formation of a HCO3- transport metabolon in exocrine cells of Nhe1-/- mice.

Cl(-) influx across the basolateral membrane is a limiting step in fluid production in exocrine cells and often involves functionally linked Cl(-)/HCO(3)(-) (Ae) and Na(+)/H(+) (Nhe) exchange mechanisms. The dependence of this major Cl(-) uptake pathway on Na(+)/H(+) exchanger expression was examined in the parotid acinar cells of Nhe1(-/-) and Nhe2(-/-) mice, both of which exhibited impaired fluid secretion. No change in Cl(-)/HCO(3)(-) exchanger activity was detected in Nhe2-deficient mice. Conversely, Cl(-)/HCO(3)(-) exchanger activity increased nearly 4-fold in Nhe1-deficient mice, despite only minimal or any change in mRNA and protein levels of the anion exchanger Ae2. Acetazolamide completely blocked the increase in Cl(-)/HCO(3)(-) exchanger activity in Nhe1-null mice suggesting that increased anion exchange required carbonic anhydrase activity. Indeed, the parotid glands of Nhe1(-/-) mice expressed higher levels of carbonic anhydrase 2 (Car2) polypeptide. Moreover, the enhanced Cl(-)/HCO(3)(-) exchange activity was accompanied by an increased abundance of Car2.Ae2 complexes in the parotid plasma membranes of Nhe1(-/-) mice. Anion exchanger activity was also significantly reduced in Car2-deficient mice, consistent with an important role of a putative Car2.Ae2 HCO(3)(-) transport metabolon in parotid exocrine cell function. Increased abundance of this HCO(3)(-) transport metabolon is likely one of the multiple compensatory changes in the exocrine parotid gland of Nhe1(-/-) mice that together attenuate the severity of in vivo electrolyte and acid-base balance perturbations.

Transcriptional regulation by targeted expression of architectural transcription factor high mobility group A2 in salivary glands of transgenic mice.

High mobility group A2 (HMGA2) protein is a non-histone architectural transcription factor. Numerous studies have demonstrated that HMGA2 is exclusively expressed in the nucleus of embryonic, but not of terminally differentiated, cells, and aberrant expression of HMGA2 is associated with various benign tumors, including pleomorphic salivary adenoma. Herein, we report the use of a 4.5-kb enhancer/promoter region of the aquaporin-5 (AQP-5) gene to target HMGA2 transgene expression in the mouse salivary acinar cells as a model to investigate the biochemical and biological role of ectopic HMGA2 expression. The expression pattern was analyzed by microarray analyses to profile HMGA2-dependent salivary gene regulation. By using quantitative reverse transcription-polymerase chain reaction (RT-PCR) assays, the expression of a cluster of genes involved in cytokine signaling, including Il7r, Il2rg, and Ptprc, was verified to be up-regulated in the salivary glands of AQP-5/HMGA2 mice. In concert, the expression of a cluster of genes, namely Ppara, Phyh, and Cidea, governing fatty acid and lipid metabolism, was confirmed to be down-regulated by HMGA2. Additionally, squamous carcinoma-like salivary tumors were observed in the AQP-5/HMGA2 transgenic mice, albeit at a low incidence. Our findings indicate that the AQP-5 promoter/enhancer-containing region is sufficient to target salivary-specific transgene expression and suggest novel roles for HMGA2 in salivary epithelial cells.

SUMOylation attenuates sensitivity toward hypoxia- or desferroxamine-induced injury by modulating adaptive responses in salivary epithelial cells.

Hypoxic stress activates various signal transduction pathways including posttranslational modification with the ubiquitin-like SUMO protein (SUMOylation). However, the molecular mechanisms by which SUMOylation regulates hypoxic responses remain unclear. Here, we investigated the ability of rat salivary Pa-4 epithelial cells to resist cell injury elicited by 1% O(2)- or hypoxia-mimetic desferroxamine (DFO)-stimulated SUMOylation processes. By using Pa-4 cells stably transduced with lenti-SUMO-1 and a cell-permeant peptide harboring SUMO-binding motif to interfere with SUMO-dependent protein-protein interactions, we demonstrate that SUMOylation augments cell survival against DFO treatment. This appeared to be partly mediated through attenuation of Protein Kinase C (PKC)-delta activation and caspase-3 cleavage, hallmarks of pro-apoptotic signaling. Intriguingly, DFO-induced phosphorylation of DNA damage marker ataxia-telangiectasia-mutated protein S1981 preceded activation of PKCdelta and caspase-3. Constitutive SUMOylation facilitated 1% O(2)- or DFO-induced nuclear factor kappaB transactivation, possibly via activation of genotoxic signaling cascade. In addition, we observed transient preservation of transepithelial electrical resistance during the early stage of hypoxia (1% O(2)) as well as enhanced transepithelial electrical resistance recovery after prolonged hypoxia in SUMO-1-expressing cell monolayers. In conclusion, our results unveil a previously unrecognized mechanism by which SUMOylation and activation of ataxia-telangiectasia-mutated protein, PKCdelta, caspase-3, and nuclear factor kappaB signaling pathways modulate salivary adaptive responses to stress in cells exposed to either 1% O(2) or DFO.

SUMOylation plays a role in gemcitabine- and bortezomib-induced cytotoxicity in human oropharyngeal carcinoma KB gemcitabine-resistant clone.

Bortezomib, a novel dipeptide boronic acid proteasome inhibitor, has been shown in previous studies to be synergistic with gemcitabine; however, the molecular mechanisms are not fully understood. Because post-translational modification of proteins, such as ubiquitination and SUMOylation, plays a critical role in governing cellular homeostasis, we explored this further by treating human oropharyngeal carcinoma KB wild-type (KBwt) and gemcitabine-resistant (KBGem) cells with gemcitabine and bortezomib in a time-dependent and sequence-dependent manner. Treatment with bortezomib at 4 to 8 hours post-gemcitabine significantly induced cell death in KBwt cell lines. However, in KBGem cells, bortezomib alone was just as cytotoxic. Using reporter assays, nuclear factor-kappaB (NF-kappaB) activity was found to be 5-fold higher in KBGem cells than that in KBwt cells, and the combination treatment decreased NF-kappaB activity by 44% in KBwt cells and 28% in KBGem cells, respectively. By Western blot analyses, treatment with gemcitabine and bortezomib resulted in a cleavage of NF-kappaB in KBwt but not in KBGem cells. SUMOylation capacity was modulated by transducing KBwt and KBGem cells with lenti-SUMO-1 or the unconjugatable lenti-SUMO-1aa followed by drug treatment. The expression of cyclins A, D1, and E was differentially regulated by SUMOylation capacity in KBGem but not in KBwt cells. We report herein that the activation of NF-kappaB signaling plays a critical role in eliciting KBwt cell survival against gemcitabine, whereas the role of SUMOylation in modulating the steady-state levels of key cell cycle regulator proteins seems more significant in KBGem cells.

Cl(-)/HCO(3)(-) exchange is acetazolamide sensitive and activated by a muscarinic receptor-induced [Ca(2+)](i) increase in salivary acinar cells.

Large volumes of saliva are generated by transepithelial Cl(-) movement during parasympathetic muscarinic receptor stimulation. To gain further insight into a major Cl(-) uptake mechanism involved in this process, we have characterized the anion exchanger (AE) activity in mouse serous parotid and mucous sublingual salivary gland acinar cells. The AE activity in acinar cells was Na(+) independent, electroneutral, and sensitive to the anion exchange inhibitor DIDS, properties consistent with the AE members of the SLC4A gene family. Localization studies using a specific antibody to the ubiquitously expressed AE2 isoform labeled acini in both parotid and sublingual glands. Western blot analysis detected an approximately 170-kDa protein that was more highly expressed in the plasma membranes of sublingual than in parotid glands. Correspondingly, the DIDS-sensitive Cl(-)/HCO(3)(-) exchanger activity was significantly greater in sublingual acinar cells. The carbonic anhydrase antagonist acetazolamide markedly inhibited, whereas muscarinic receptor stimulation enhanced, the Cl(-)/HCO(3)(-) exchanger activity in acinar cells from both glands. Intracellular Ca(2+) chelation prevented muscarinic receptor-induced upregulation of the AE, whereas raising the intracellular Ca(2+) concentration with the Ca(2+)-ATPase inhibitor thapsigargin mimicked the effects of muscarinic receptor stimulation. In summary, carbonic anhydrase activity was essential for regulating Cl(-)/HCO(3)(-) exchange in salivary gland acinar cells. Moreover, muscarinic receptor stimulation enhanced AE activity through a Ca(2+)-dependent mechanism. Such forms of regulation may play important roles in modulating fluid and electrolyte secretion by salivary gland acinar cells.

Secretion and cell volume regulation by salivary acinar cells from mice lacking expression of the Clcn3 Cl- channel gene.

Salivary gland acinar cells shrink when Cl(-) currents are activated following cell swelling induced by exposure to a hypotonic solution or in response to calcium-mobilizing agonists. The molecular identity of the Cl(-) channel(s) in salivary cells involved in these processes is unknown, although ClC-3 has been implicated in several tissues as a cell-volume-sensitive Cl(-) channel. We found that cells isolated from mice with targeted disruption of the Clcn3 gene undergo regulatory volume decrease in a fashion similar to cells from wild-type littermates. Consistent with a normal regulatory volume decrease response, the magnitude and the kinetics of the swell-activated Cl(-) currents in cells from ClC-3-deficient mice were equivalent to those from wild-type mice. It has also been suggested that ClC-3 is activated by Ca(2+)-calmodulin-dependent protein kinase II; however, the magnitude of the Ca(2+)-dependent Cl(-) current was unchanged in the Clcn3(-/-) animals. In addition, we observed that ClC-3 appeared to be highly expressed in the smooth muscle cells of glandular blood vessels, suggesting a potential role for this channel in saliva production by regulating blood flow, yet the volume and ionic compositions of in vivo stimulated saliva from wild-type and null mutant animals were comparable. Finally, in some cells ClC-3 is an intracellular channel that is thought to be involved in vesicular acidification and secretion. Nevertheless, the protein content of saliva was unchanged in Clcn3(-/-) mice. Our results demonstrate that the ClC-3 Cl(-) channel is not a major regulator of acinar cell volume, nor is it essential for determining the secretion rate and composition of saliva.

Loss of hyperpolarization-activated Cl(-) current in salivary acinar cells from Clcn2 knockout mice.

ClC-2 is localized to the apical membranes of secretory epithelia where it has been hypothesized to play a role in fluid secretion. Although ClC-2 is clearly the inwardly rectifying anion channel in several tissues, the molecular identity of the hyperpolarization-activated Cl(-) current in other organs, including the salivary gland, is currently unknown. To determine the nature of the hyperpolarization-activated Cl(-) current and to examine the role of ClC-2 in salivary gland function, a mouse line containing a targeted disruption of the Clcn2 gene was generated. The resulting homozygous Clcn2(-/-) mice lacked detectable hyperpolarization-activated chloride currents in parotid acinar cells and, as described previously, displayed postnatal degeneration of the retina and testis. The magnitude and biophysical characteristics of the volume- and calcium-activated chloride currents in these cells were unaffected by the absence of ClC-2. Although ClC-2 appears to contribute to fluid secretion in some cell types, both the initial and sustained salivary flow rates were normal in Clcn2(-/-) mice following in vivo stimulation with pilocarpine, a cholinergic agonist. In addition, the electrolytes and protein contents of the mature secretions were normal. Because ClC-2 has been postulated to contribute to cell volume control, we also examined regulatory volume decrease following cell swelling. However, parotid acinar cells from Clcn2(-/-) mice recovered volume with similar efficiency to wild-type littermates. These data demonstrate that ClC-2 is the hyperpolarization-activated Cl(-) channel in salivary acinar cells but is not essential for maximum chloride flux during stimulated secretion of saliva or acinar cell volume regulation.