Development of an ectopic huLiver model for Plasmodium liver stage infection.

Early Plasmodium falciparum and P. vivax infection requires parasite replication within host hepatocytes, referred to as liver stage (LS). However, limited understanding of infection dynamics in human LS exists due to species-specificity challenges. Reported here is a reproducible, easy-to-manipulate, and moderate-cost in vivo model to study human Plasmodium LS in mice; the ectopic huLiver model. Ectopic huLiver tumors were generated through subcutaneous injection of the HC-04 cell line and shown to be infectible by both freshly dissected sporozoites and through the bite of infected mosquitoes. Evidence for complete LS development was supported by the transition to blood-stage infection in mice engrafted with human erythrocytes. Additionally, this model was successfully evaluated for its utility in testing antimalarial therapeutics, as supported by primaquine acting as a causal prophylactic against P. falciparum. Presented here is a new platform for the study of human Plasmodium infection with the potential to aid in drug discovery.

Morphological alteration of the pancreatic islet in ovariectomized rats fed a high-fat high-fructose diet.

Diabetes and its complications are major causes of mortality worldwide. Type 2 diabetes coexists with insulin resistance and ß-cell dysfunction, which are aggravated by overconsumption and estrogen-deprived conditions. However, the morphology of pancreatic islets in a combined condition of excessive caloric intake and estrogen deficiency has never been described. Herein, we examined morphological changes in the pancreatic islets of ovariectomized (OVX) rats fed a high-fat high-fructose diet (HFFD) for 12 weeks. The histological changes in the size and number of pancreatic islets were assessed by hematoxylin-eosin and immunohistochemical staining. Enlarged pancreatic islets with fat deposition in OVX rats were accompanied by whole-body insulin resistance and hyperglycemia. The addition of a HFFD to OVX rats (OVX + HFFD) further aggravated insulin resistance, with a substantial increase in the density of enlarged pancreatic islets and fat accumulation. The augmented number of enlarged islets was correlated with elevated plasma glucose and insulin levels. Intriguingly, unlike the HFFD and OVX alone, the OVX + HFFD markedly expanded the area of insulin-producing ß-cells and glucagon-producing α-cells. Importantly, enlarged islets, pancreatic fat deposits, and diabetic states developing in OVX + HFFD conditions were resolved by estrogen replacement. Collectively, the morphological characteristics of pancreatic islets were influenced in an insulin-resistant state caused by estrogen deficiency and HFFD consumption and were distinct from each factor alone. A combination of estrogen deficiency with HFFD consumption worsened the integrity of pancreatic islets, ultimately resulting in disease progression. These findings expand our understanding of the causal relationship between pancreatic morphology and diabetes development and suggest therapeutic strategies.

Efficacy of Eosin Dye Removal by Peanut Shell Agrowaste Adsorbent

Abstract Peanut shell (PS) which is an excessive waste-product from agricultural processes, it can be recycled to a natural adsorbent for example it uses as removal dyes. Synthetic dye effluent without improperly discharged from industries to the river cause wastewater and damage to living organisms, especially, anionic dyes are difficult removed by conventional treatments such as biological, chemical, oxidation, and physical-filtration. However, an adsorption treatment is widely used for decolorization of dyes and give the best results for removal of various types of dissolved coloring materials. This research was used Eosin Y (EO) for the anionic model of dyeing wastewater and used PS for agrowaste adsorbent. The purpose of this study was investigated the efficiency adsorption of EO removal by PS. This efficiency adsorption was measured by different PS dosages, contact times, adsorbate concentration and equilibrium data. The results can be concluded that the PS had the efficiency adsorption of EO removal due to the equilibrium adsorption capacity (qe) and the highest dose of PS were balanced to adsorption of dye. The highest EO removal percentage was found in 87.7%, the qe was 0.351 mg g-1 and can adsorb from 10 mg L-1 to 1.23 mg L-1 in 25 g L-1 of PS dose at 30 minutes. In addition, the PS structure was found in multi-layer and many porous which is suitable for adsorbent. The morphological examination of PS was shown before and after adsorption that not changed. Therefore, PS might be an alternative choice for removal dye, and be used for the recycle adsorbent agrowaste as a commercial product for adding their values.

The sbGnRH-GTH system in the female short mackerel, Rastrelliger brachysoma (Bleeker, 1851), during breeding season: implications for low gamete production in captive broodstock.

The short mackerel (Rastrelliger brachysoma) is one of the most economically important fish in Thailand; it is also a prime candidate for mariculture but unfortunately is plagued by reproductive problems that cause low production of gametes in captivity. An understanding of how the brain, pituitary, and gonad axis (BPG) from the neuroendocrine system are involved in the reproductive activity of wild and captive R. brachysoma should help clarify the situation. In this study, we investigated changes in the sea bream gonadotropin-releasing hormone (sbGnRH)-gonadotropin (GTH) system in the female short mackerel, Rastrelliger brachysoma (Bleeker, 1851), during breeding season. sbGnRH-immunoreactive (ir) cell bodies were detected in the nucleus preopticus-periventricularis including nucleus periventricularis (NPT), nucleus preopticus (Np), and nucleus lateralis tuberis (NLT). Additionally, the sbGnRH-ir fibers protruded into the proximal par distalis (PPD) where GTH (FSH and LH) cells were detected. The number of sbGnRH-ir cell bodies and GTH cells and level of LH mRNA were significantly higher in the breeding season than those in the non-breeding season. Moreover, the number of sbGnRH-ir cell bodies and GTH cells and levels of sbGnRH and GTH (FSH and LH) mRNA were significantly higher in the wild fish than those in the cultured broodstock. It is suggested that the wild fish tended to have better reproductive system than hatchery fishes. This could be related to the endocrinological dysfunction and the reproductive failure in the hatchery condition. Moreover, the changes of all of the hormonal level could potentially be applied to R. brachysoma aquaculture.

Controlled rapid synthesis and in vivo immunomodulatory effects of LM α(1,6)mannan with an amine linker.

The synthetic lipomannan (LM) α(1,6)mannans, already equipped with an amine linker on the reducing end, are rapidly synthesized in a size-, regio-, and stereocontrolled reaction. The size of the mannans is regulated through the concentration of the linker, applied during the controlled ring-opening polymerization reaction. The versatile amine linker enables a variety of glycan conjugations. The synthetic α(1,6)mannans exert adjuvant activities for a real vaccine antigen, tetanus toxoid (TT) in vitro, as demonstrated by the increased secretion of proinflammatory cytokines TNF-α and IL-6 from the treated macrophages. A conjugation of synthetic α(1,6)mannan with TT can also enhance immune response to TT in vivo after immunization as shown by an increase in TNF-α, IFN-γ, and IL-2 production in splenocytes.

Synthesis and Immunological Studies of the Lipomannan Backbone Glycans Found on the Surface of Mycobacterium tuberculosis.

Investigations into novel bacterial drug targets and vaccines are necessary to overcome tuberculosis. Lipomannan (LM), found on the surface of Mycobacterium tuberculosis (Mtb), is actively involved in the pathogenesis and survival of Mtb. Here, we report for the first time a rapid synthesis and biological activities of an LM glycan backbone, α(1-6)mannans. The rapid synthesis is achieved via a regio- and stereoselective ring opening polymerization to generate multiple glycosidic bonds in one simple chemical step, allowing us to finish assembling the defined polysaccharides of 5-20 units within days rather than years. Within the same pot, the polymerization is terminated by a thiol-linker to serve as a conjugation point to carrier proteins and surfaces for immunological experiments. The synthetic glycans are found to have adjuvant activities in vivo. The interactions with DC-SIGN demonstrated the significance of α(1-6)mannan motif present in LM structure. Moreover, surface plasmon resonance (SPR) showed that longer chain of synthetic α(1-6)mannans gain better lectin's binding affinity. The chemically defined components of the bacterial envelope serve as important tools to reveal the interactions of Mtb with mammalian hosts and facilitate the determination of the immunologically active molecular components.

Plasmodium vivax liver stage development and hypnozoite persistence in human liver-chimeric mice.

Plasmodium vivax malaria is characterized by periodic relapses of symptomatic blood stage parasite infections likely initiated by activation of dormant liver stage parasites-hypnozoites. The lack of tractable P. vivax animal models constitutes an obstacle in examining P. vivax liver stage infection and drug efficacy. To overcome this obstacle, we have used human liver-chimeric (huHep) FRG KO mice as a model for P. vivax infection. FRG KO huHep mice support P. vivax sporozoite infection, liver stage development, and hypnozoite formation. We show complete P. vivax liver stage development, including maturation into infectious exo-erythrocytic merozoites as well as the formation and persistence of hypnozoites. Prophylaxis or treatment with the antimalarial primaquine can prevent and eliminate liver stage infection, respectively. Thus, P. vivax-infected FRG KO huHep mice are a model to investigate liver stage development and dormancy and may facilitate the discovery of drugs targeting relapsing malaria.

Schistosoma mekongi: the in vitro effect of praziquantel and artesunate on the adult fluke.

The efficacy and tolerance of 80 microg/ml praziquantel (PZQ) and 40 microg/ml artesunate (ATS) against adult stage Schistosoma mekongi in vitro were investigated after 3, 6, 12, and 24h incubation by monitoring worm motility and compared tegumental changes using scanning electron microscopy (SEM). Thirty mice were infected with S. mekongi cercaria for 49 days. Adult worms were collected by perfusion method and prepared for in vitro study. Contraction and decreased motor activity were observed after as little as 3h incubation with PZQ and ATS. Some of the worms were immobile 12h after exposure, and died within 24h. The tegument of S. mekongi showed severe swelling, vacuolization and disruption, fusion of the tegumental ridges, collapse and peeling. After 12-24h incubation, PZQ induced similar but they less severe, tegumental changes to those observed after exposure to ATS. The direct observation of the fluke motility and SEM study suggest that ATS is more effective than PZQ in causing tegumental damage in adult S. mekongi, and provides a basis for subsequent clinical trials.

Eurytrema pancreaticum: the in vitro effect of praziquantel and triclabendazole on the adult fluke.

The efficacy and tolerance of the 80 microg/ml praziquantel (PZQ) and 40 microg/ml triclabendazole (TCZ) against adult stage Eurytrema pancreaticum in vitro were investigated at 3, 12, and 15 h incubation. Motility of the flukes and histopathological changes were studied. Sudden paralysis and death were observed after exposed to PZQ as early as 3h incubation. In contrast, the TCZ treated flukes showed active mobility at all intervals. By light microscopic examination, severe damages in various organs such as tegument, muscle, and testes were observed early at 12h incubation of these drugs. PZQ caused more severe damage to flukes than TCZ. There were vigorous contraction of musculature, progressive shrinkage of circular and longitudinal muscles, vacuolization and disintegration of the tegument disrupting the worms' outer surface including detachment of spines in the PZQ treatment. The cells in testes were slightly increased in size and followed by degeneration leaving several hollow spaces. The uterus and vitelline glands remained unaffected. The direct observation of the fluke motility and light microscopic study highly suggested that PZQ was more effective than TCZ treatment for the eurytremiasis infection.

Effects of praziquantel and artesunate on the tegument of adult Schistosoma mekongi harboured in mice.

The effects of praziquantel and artesunate on the tegument of adult Schistosoma mekongi harboured in mice were compared using scanning electron microscopy (SEM). Forty-two mice infected with S. mekongi for 49 days were treated intragastrically with either 300 mg/kg praziquantel or 300 mg/kg artesunate. Mice were sacrificed 1 or 3 days post-treatment. Worms were collected by perfusion and examined by SEM. One to 3 days after administration of artesunate, the tegument of S. mekongi showed severe swelling, vacuolization, fusion of the tegumental ridges and loss or shortening of the spines on the trabeculae, collapse and peeling. Praziquantel induced similar tegumental alterations as those observed after administration of artesunate, but they were less severe. Three days post-treatment, there was evidence of recovery only in the case of praziquantel. The results of our study suggest that artesunate is more effective than praziquantel in causing tegumental damage in adult S. mekongi, and provides a basis for subsequent clinical trials.