IL-33 Enhances the Total Production of IgG, IgG1, and IgG3 in Angiostrongylus cantonensis-Infected Mice.

The purpose of this study is to clarify the role of IL-33 in the immune response to angiostrongyliasis, especially in terms of antibody production and isotype switching. In our experiment, C57BL/6 mice were each infected with 35 infectious larvae and were divided into groups that received an intraperitoneal injection of IL-33, anti-IL-33 monoclonal antibody (mAb), or anti-ST2 mAb 3 days post-infection (dpi) and were subsequently administered booster shots at 5-day intervals with the same dose. Serum samples from each group were collected weekly for ELISA assays. The levels of total IgG, IgG1, and IgG3 were significantly increased in A. cantonensis-infected mice that were treated with IL-33, and the levels decreased significantly in infected groups treated with anti-IL-33 or anti-ST2 mAb. These results suggest that IL-33 may play a critical role in the pathogenesis of human angiostrongyliasis and could be useful for understanding protective immunity against this parasitic infection.

Hypothermia on the first day of ICU admission leads to increased in-hospital mortality in patients with subarachnoid hemorrhage.

The relationship between early spontaneous hypothermia and adverse clinical outcomes in patients with subarachnoid hemorrhage (SAH) has not been paid much attention. We designed this retrospective cohort study to determine this relationship by analyzing the association between the lowest body temperature (T-lowest) on the first day of ICU admission and in-hospital mortality. In this study, 550 participants with non-traumatic SAH were chosen from the Medical Information Mart for Intensive Care (MIMIC)-IV database. Multivariate Cox regression analysis showed that T-lowest was nonlinearity correlated with in-hospital mortality (HR = 0.72, 95% CI: 0.59-0.86, p < 0.001). We divided the T-lowest into quartile groups. In comparison to reference group Q1 (31.30-36.06 â„ƒ), group Q3 (36.56-36.72 â„ƒ) had a 50% lower risk of death in the hospital (HR: 0.5, 95% CI: 0.28-0.87, p = 0.014). We further confirmed the curve-like relationship between T-lowest and in-hospital mortality using restricted cubic splines. The mortality is lowest when the T-lowest is close to 36.5 °C, and the risk of death is increased when the temperature is lower or higher than that. Our study demonstrates that in-hospital mortality is associated with T-lowest. Patients with non-traumatic SAH are at increased risk of death if their body temperature on the first day of ICU admission is too low.

Relationship between baseline bicarbonate and 30-day mortality in patients with non-traumatic subarachnoid hemorrhage.

Objective: This study aimed to explore the relationship between baseline bicarbonate levels and 30-day mortality in individuals with non-traumatic subarachnoid hemorrhage (SAH). Methods: Patients with non-traumatic SAH were chosen from the Medical Information Mart for Intensive Care (MIMIC)-IV database. The relationship between baseline bicarbonate and 30-day mortality was examined using Cox regression models. Restricted cubic splines were used to test the hypothesis that there was an association between bicarbonate and mortality. With the use of Kaplan-Meier survival curve analysis, we looked deeper into the validity of these correlations. To find subgroups with differences, interaction tests were utilized. Results: This retrospective cohort study consisted of 521 participants in total. Bicarbonate had a negative association with death at 30 days (HR = 0.93, 95%CI: 0.88-0.98, p = 0.004). Next, we divided bicarbonate into quartile groups. In comparison to the reference group Q1 (20 mEq/L), groups Q3 (23-25 mEq/L) and Q4 (26 mEq/L) had adjusted HR values of 0.47 (95%CI: 0.27-0.82, p = 0.007) and 0.56 (95%CI: 0.31-0.99, p = 0.047). No definite conclusions can be derived from this study, since there is no obvious curve link between baseline bicarbonate and 30-day mortality. Patients' 30-day mortality increased statistically significantly (p < 0.001, K-M analysis) in patients with low bicarbonate levels. The relationship between bicarbonate and 30-day mortality remained consistent in the stratified analysis, with no observed interactions. Conclusion: Finally, 30-day mortality was negatively associated with baseline bicarbonate levels. Patients with non-traumatic SAH are more at risk of mortality if their bicarbonate levels are low.

Chlorogenic acid in the oxidative stress injury triggered by Shuang-Huang-Lian injection.

Injections of Chinese herbs are a novel approach to prepare traditional Chinese medicines. However, as injections of Chinese herbs have been extensively used, adverse drug reactions (ADRs) have been on the increase. Additionally, the mechanism for injections of Chinese herbs remains unclear. This study explored the potential role played by chlorogenic acid (CGA) in initiating oxidative stress injury triggered by the utilization of injections of Chinese herbs and the underlying mechanism. A total of 90 male Wistar rats were raised for varying periods by using Shuang-Huang-Lian (SHL) injection or CGA in diverse dosages. Western blot analysis examined the expression of nicotinamide adenine dinucleotide phosphate oxidase subunits, spectrophotometry was used to examine the activity taken by catalase, ELISA was used to examine the concentrations of inflammatory factors in serum, and intravital microscopy was employed to examine the microcirculation. The results showed that the excessive peroxide production induced by CGA in high-dose or SHL in the venule walls may well be through nicotinamide adenine dinucleotide phosphate oxidase along with a decline in the activity of catalase, and led to imbalance of basal levels of pro-(TNF-α) and anti-(IL-10) inflammatory cytokines. On the basis of the aforementioned results, the mechanism hidden behind the adverse effects of CGA induced by irrational use of Chinese herbal injection can be identified from a deeper perspective.

Involvement of NADPH oxidase in high-dose phenolic acid-induced pro-oxidant activity on rat mesenteric venules.

In the present study, we investigated the potential role of phenolic acids in initiating oxidative damage to microvascular endothelial cells and the underlying mechanism mediating the pro-oxidant action. Male Wistar rats received high doses of phenolic acid [caffeic acid (CA), salvianolic acid B (SAB), chlorogenic acid (ChA) or ferulic acid (FA)]. The creation of reactive oxygen species in mesenteric microcirculation endothelial cells and adherent leukocytes along with venules were assessed using intravital microscopy. The expression levels of NADPH oxidase subunits (Nox4 and p22phox) in terminal ileum tissues were determined by western blot analysis. Intravenous injection of high-dose ChA or CA (7 mg/kg) markedly increased the peroxide production in the venular walls and upregulated the protein expression levels of Nox4 and p22phox in the ileum tissues, while the same dose of CA and SAB made no difference within the observation period. No changes were observed in the number of leukocytes adhering to the venular walls. High-dose ChA and FA led to an imbalance between the oxidant and antioxidant mechanism by boosting the expression levels of NADPH oxidase. Thus, we clarified the rationale behind the adverse effects of a herbal injection containing high levels of phenolic acid compounds.

Anti-ST2 monoclonal antibody inhibits eosinophil infiltration in Angiostrongylus cantonensis-infected mice.

BACKGROUND/PURPOSE: Interleukin-33 (IL-33) could play an important role in the pathogenesis of angiostrongylosis. However, the role of IL-33/ST2 pathway in this parasitic infection is uncertain. METHODS: C57BL/six mice were each infected with 35 Angiostrongylus cantonensis larvae. One group of mice received an intraperitoneal injection of anti-ST2 monoclonal antibody (mAb; 50 µg) 3 days postinfection and subsequent booster shots of the same dose at 5-day intervals. Blood samples from each group were collected every week for assays. RESULTS: The level of IL-5 significantly decreased in the mAb-treated group, and the infiltration of eosinophils in the meninges was also significantly reduced. CONCLUSION: The IL-33/ST2 axis may play a crucial role in the pathogenesis of angiostrongylosis and the results of this study could be useful for the development of strategies to reduce the neurological damage caused by this parasitic infection.

Signal acquisition module design for multi-channel surface magnetic resonance sounding system.

To obtain a precise 2D/3D image of fissure or karst water, multi-channel magnetic resonance sounding (MRS) systems using edge-to-edge or overlapping receiving coils are needed. Thus, acquiring a nano-volt signal for a small amount of the aquifer and suppressing the mutual coupling between adjacent coils are two important issues for the design of the signal acquisition module in the system. In the present study, we propose to use a passive low pass filter, consisted of a resistance (R) and capacitance (C), to inhibit the mutual coupling effects of the coils. Four low-noise operational amplifiers LT1028, OPA124, AD745, and OP27 were compared with respect to achieving the lowest system noise. As a result, 3 pieces of LT1028 were chosen and connected in parallel to serve as preamplifier, with a sensitivity of 1.4 nV/√Hz at 2 kHz. Experimental results are presented for 2D MRS groundwater investigations conducted in the suburb of Changchun, China. The inversion result is consistent with the result of drilling log, suggesting that the signal acquisition module is well developed.

IL-33 mediates the expressions of IL-5 and IL-13 in Angiostrongylus cantonensis-infected mice.

Angiostrongylus cantonensis is the major cause of human eosinophilic meningoencephalitis. C57BL/6 mice were experimentally infected with 35 infectious larvae. Two groups of infected mice received intraperitoneal injections of mouse IL-33 (1µg) or anti-IL-33 monoclonal antibody (mAb) (10µg) 3days post infection (dpi) and subsequent booster shots of the same dose at 5day intervals. Blood samples from each group were collected weekly for assays. IgE levels were significantly increased in all infected mice. The eosinophil percentage and levels of IL-5 and IL-13 significantly increased in the IL-33-treated group relative to infected but non-treated animals. The level of IL-5 decreased in the mAb-treated group. The severity of eosinophilic meningitis was exacerbated in the IL-33 injected group. Taken together, these results suggest that IL-33 mediates the expressions of IL-5 and IL-13, and plays a crucial role in the pathogenesis of angiostrongylosis.

Amelioration of superoxide dismutase on ventilator-induced lung injury by suppressing leukocyte in the lungs and systemic circulation.

Superoxide dismutase (SOD) is a free radical scavenger and a broad-spectrum antioxidant. Its anti-inflammatory and immunomodulatory effects have recently been noted. We studied the effects of this antioxidant on lung damage, oxidative stress, and inflammation in a model of ventilator-induced lung injury (VILI), using 8- to 12-wk-old Sprange-Dawley rats (n = 40). Animals were randomized and evenly divided into two experimental groups, low tidal volume (V(T)) ventilation (V(T) = 9 ml/kg) and high V(T) ventilation (V(T) = 28 ml/kg). Each group was evenly divided into two subgroups: ten animals were treated with superoxide dismutase (SOD; 10,000 U/kg i.v., 2 h prior to the ventilation) and the rests were treated with vehicle. Lung injury was evaluated by histological examination, and cells counts of red blood cells (RBC) and white blood cells (WBC) in the alveoli and the septal wall thickness in lung tissues and serum lactate dehydrogenase (LDH). The lung permeability was assessed by the wet-to-dry weight ratio (W/D), lung weight to body weight ratio (LW/BW) and protein concentration in broncholavage fluid (BALF). Levels of oxidative stress and lipid peroxidation in the lungs were evaluated by tissue malondialdehyde (MDA) and methylguanidine (MG) in BALF, respectively. SOD pretreatment significantly decreased WBC counts in systemic circulation and in alveoli, and effectively attenuated high V(T) ventilation induced lung injury by reducing hyaline membrane development, septal wall thickness, lung W/D and LW/BW and serum LDH in relation to those of the control. In addition, lung tissues MDA and MG in BALF were also notably reduced.

A RAB family antigen of Angiostrongylus cantonensis induces a Th1-type immune response in vitro and in vivo.

A cDNA library was constructed from an Angiostrongylus cantonensis young adult and the encoded proteins were expressed in Escherichia coli. One reactive antigen, a RAB-2 protein, was selected using an immunoscreening technique. The expression of the Th1-type cytokine IFN-γ was elicited in mouse splenic cells that were co-cultured with the recombinant RAB-2 protein and in the sera of mice that were immunised with this protein and adjuvant (50 µg at 2-week intervals). In the A. cantonensis-infected groups, the mice were orally infected with 35 infective larvae, and a subset of the infected mice were immunised with the recombinant RAB-2 protein in adjuvant. Serum samples were collected every week for ELISA, and the pathological examinations were performed at 14 days post infection (dpi). An increase in IFN-γ expression was noted in the blood, and the brain sections revealed moderate eosinophilic meningitis in the immunised mice. The RAB-2 antigen of A. cantonensis induced a Th1-type immune response both in vitro and in vivo.

High-dose chlorogenic acid induces inflammation reactions and oxidative stress injury in rats without implication of mast cell degranulation.

ETHNOPHARMACOLOGICAL RELEVANCE: Chlorogenic acid (CA) exits widely in those Chinese herbal injections that have antibacterial and antiphlogistic effects and belongs to the ethnopharmacological family of medicines. Chinese herbal injections containing high levels of CA have been reported to increase the adverse drug reactions, but the mechanism for which is still unclear. In this study, we investigated the mechanism of the CA derived adverse drug reactions. AIM OF THE STUDY: The present study was to explore the potential role of CA in initiating inflammatory reaction and the underlying mechanism. MATERIALS AND METHODS: Male Wistar rats were treated with different dosages of CA for different time period. The variables examined included microcirculation by intravital microscopy, histology of ileum tissue, expression of adhesion molecules CD11b and CD18 on leukocytes by flow cytometry, myeloperoxidase activity and maleic dialdehyde content in ileum tissue by spectrophotometry, activity of superoxide dismutase and catalase in serum by ELISA, and expression of NADPH oxidase subunits by PCR and Western blot. RESULTS: High-dose CA increased the number of adherent leukocytes, generation of peroxides in the venular walls and induced albumin leakage from mesentery venules. High-dose CA induced changes also included an increase in maleic dialdehyde, myeloperoxidase, inflammatory cytokines and NADPH oxidase activities, and a decline in activity of superoxide dismutase and catalase. CONCLUSION: High-dose, but not Low-dose CA induced inflammation reaction, and in this process an imbalance between oxidant and antioxidant mechanism may be involved, providing more information for better understanding the rationale behind the adverse effects of CA.

A naturally occurring carotenoid, lutein, reduces PDGF and H2O2 signaling and compromised migration in cultured vascular smooth muscle cells.

BACKGROUND: Platelet-derived growth factor (PDGF) is a potent stimulator of growth and motility of vascular smooth muscle cells (VSMCs). Abnormalities of PDGF/PDGF receptor (PDGFR) are thought to contribute to vascular diseases and malignancy. We previously showed that a carotenoid, lycopene, can directly bind to PDGF and affect its related functions in VSMCs. In this study we examined the effect of the other naturally occurring carotenoid, lutein, on PDGF signaling and migration in VSMCs. METHODS: Western blotting was performed to examine PDGF and H2O2 signaling. Flowcytometry was used to determine PDGF binding to VSMCs. Fluorescence microscopy was performed to examine intracellular ROS production. Modified Boyden chamber system (Transwell apparatus) was used for migration assay. RESULTS: Lutein reduced PDGF signaling, including phosphorylation of PDGFR-ß and its downstream protein kinases/enzymes such as phospholipase C-γ, Akt, and mitogen-activated protein kinases (MAPKs). Although lutein possesses a similar structure to lycopene, it was striking that lutein inhibited PDGF signaling through a different way from lycopene in VSMCs. Unlike lycopene, lutein not only interacted with (bound to) PDGF but also interfered with cellular components. This was evidenced that preincubation of PDGF with lutein and treatment of VSMCs with lutein followed by removing of lutein compromised PDGF-induced signaling. Lutein reduced PDGF-induced intracellular reactive oxygen species (ROS) production and attenuated ROS- (H2O2-) induced ERK1/2 and p38 MAPK activation. A further analysis indicated lutein could inhibit a higher concentration of H2O2-induced PDGFR signaling, which is known to act through an oxidative inhibition of protein tyrosine phosphatase. Finally, we showed that lutein functionally inhibited PDGF-induced VSMC migration, whereas its stereo-isomer zeaxanthin did not, revealing a special action of lutein on VSMCs. CONCLUSIONS: Our study reveals a differential action mechanism of lutein from other reported caroteinoids and suggests a possible beneficial effect of lutein but not zeaxanthin on prevention of vascular diseases.

MicroRNAs and cataracts: correlation among let-7 expression, age and the severity of lens opacity.

BACKGROUND: Let-7 microRNA is an important regulator of cellular ageing and tissue senescence. The objective of this study is to evaluate the expression of let-7a/let-7b/let-7c microRNAs in human age-related cataracts. AIM: To evaluate the correlation among the severity of lens opacity, the level of let-7a/let-7b/let-7c microRNA expression and patient age in the context of age-related cataracts. METHODS: The authors evaluated the mRNA level of let-7a/let-7b/let-7c microRNA in lens epithelia obtained from 174 eyes with age-related cataracts. The authors also recorded the patient age and the severity of lens opacity as classified according to the modified version of the Lens Opacities Classification System version III. RESULTS: Let-7b microRNA expression was demonstrated to be positively associated with patient age (R=0.472; p<0.001). A positive correlation was also observed between higher N, C and P cataract scores and higher expression of let-7b microRNA in patients with age-related cataracts (p<0.001). However, no significant correlation was observed between the let-7a and let-7c microRNA expression levels and either the severity of lens opacity or the patient age. CONCLUSION: These findings suggest that microRNAs play a role in age-related cataracts. A local let-7b microRNA increase may represent a risk factor in the formation of age-related cataracts.

Infectivity and development of Metagonimus yokogawai in experimentally infected domestic ducks (Cairina moschata).

Information concerning whether fowl such as duck is a suitable reservoir host of Metagonimus yokogawai is largely unclear to date. In the present study, the growth and development of M. yokogawai metacercaria (Mc) in domestic duck (Cairina moschata) was determined by worm recovery rate (WRR) and morphological changes e.g., the size of fluke's body as well as their internal organs was assessed by using Semichon's acetocarmine staining. Each duck was orally inoculated with 50 Mcs of M. yokogawai and infected ducks were deeply anesthetized with ether and killed at 1, 2, 3, 4, 5, 6, 7, and 14 days post-infection (DPI). On each date, two infected ducks were killed and the small intestines of each duck were separated into four parts then they were opened longitudinally to harvest the flukes. Results revealed that WRR of M. yokogawai from inoculated ducks increased during early infection with a peak as seen at 4 DPI (28.5+/-6.9%); thereafter it gradually decreased and a drastic decline was observed in 14 DPI (2.0+/-1.1%) in the trial. The preference sites for M. yokogawai were low portions of the small intestine; nevertheless the size of fluke's body and organs developed increasingly with time and they maturated to produce ova from 4 DPI onward in the trial. However, present results indicated that ducks, based on the findings of this study, are not suitable hosts for establishment of M. yokogawai infection because most flukes were expelled from duck's intestine within 14 days. Nevertheless, it was proposed that ducks might play a certain role in transmitting M. yokogawai when they deposited the ova via feces into marsh where snails and fish were abundant since they could presumably establish transient and possibly patent infections with this parasite.

Anti-CCR3 monoclonal antibody inhibits eosinophil infiltration in Angiostrongylus cantonensis-infected ICR mice.

ICR mice were each infected with 35 Angiostrongylus cantonensis larvae. One group of mice received an intraperitoneal injection of anti-CCR3 monoclonal antibody (mAb) (50 microg) at 10 days post-infection (dpi), while another similarly-treated group also received a booster injection (25 microg) at 12 dpi. All the mice were sacrificed at 14 dpi for pathological examination, enzyme-linked immunosorbent assay analysis and RNA extraction. The infiltration of eosinophils and the severity of eosinophilic meningitis were reduced in both the mAb-treated groups, relative to infected but untreated animals. The levels of CCL11 (eotaxin) in the peripheral circulation and the expression of the Th2-type cytokine interleukin-5 in the brains were significantly reduced. A. cantonensis infection is the major cause of eosinophilic meningoencephalitis in Taiwan, and the results of this study could be useful for the development of strategies to reduce the neurological damage caused by this infection.

Angiostrongylus cantonensis: apoptosis of inflammatory cells induced by treatment with mebendazole or/and interleukin 12 in mice.

Angiostrongylus cantonensis is the major cause of human eosinophilic meningoencephalitis. ICR mice were infected orally with 35 infective larvae and sacrificed at 4-14 days, 25 days or 32 days post infection (dpi) for pathological and immunocytochemical examinations. In the non-treated group, no apoptosis signal was found in the meninges or parenchyma of the brains (4-14 dpi). Only a few apoptotic cells were noticed at 25 dpi (3%) and 32 dpi (10%). In the groups, the animals were given a single dose of mebendazole (20 mg/kg, per os at various times) or injections of interleukin 12 (IL-12) (10 ng/daily, intraperitoneally), all the animals were sacrificed at 14 dpi; the number of apoptotic cells was increased (17-21%). In the group that received a single dose of mebendazole (4 dpi) in combination with IL-12 injections (4-13 dpi), mild meningitis was observed, and most of the infiltrated inflammatory cells were in the apoptotic program (55%). Taken together, apoptosis of the inflammatory cells (most were eosinophils) could be induced when the infected mice were treated with mebendazole or/and IL-12.

Seroepidemiology of Toxocara canis infection among mountain aboriginal schoolchildren living in contaminated districts in eastern Taiwan.

We conducted a seroepidemiological study of Toxocara canis infection among mountain aboriginal schoolchildren aged 7-12 years living in contaminated districts in eastern Taiwan. To detect sera IgG (> or =1:64) we used a T. canis larval excretory-secretory antigen-based enzyme-linked immunosorbent assay. A short questionnaire elicited information on the practices of raising dogs, playing with soil, eating raw vegetables, or whether the child normally washed his/her hands before eating. The overall seroprevalence was quite high, reaching 76.6% (252/329). Neither age nor gender seemed to be important factors related to a positive serology. Aboriginal schoolchildren who raised dogs (OR=1.83, 95% CI: 1.04-3.19, P=0.03), or played with soil (OR=2.52, 95% CI: 1.49-4.25, P <0.001) seemed to be more susceptible to T. canis infection than those who did not. Children who habitually washed their hands before eating (OR=0.57, 95% CI: 0.33-0.97, P=0.04) had a lower chance of acquiring T. canis infection than those who did not.

Seroepidemiology of Toxocara canis infection among mountain aboriginal adults in Taiwan.

Seroepidemiology of Toxocara canis infection among adults of one ethnic Han and five aboriginal populations residing in mountainous areas of Taiwan was conducted by detecting serum IgG (>/=1:64) using a T. canis larval excretory-secretory antigen-based enzyme-linked immunosorbent assay. A short questionnaire interview was conducted to obtain data concerning their age, sex, occupation, consumption of raw pig liver, and possession of dogs. The overall seroprevalence (46.0%, 247 of 537) in the five aboriginal populations was significantly higher than that of ethnic Han population (30.2%, 13 of 43) (P = 0.04). Age, but not sex, seemed to be a factor related to positive serology. Aboriginal adults who had histories of eating raw pig liver (odds ratio [OR] = 1.65, P < 0.01), raising dogs (OR = 1.76, P < 0.01), or whose occupation was a laborer (OR = 1.78, P < 0.01) seemed to be more apt to be infected by T. canis than those without such histories and unemployed persons.

Combined treatment with interleukin-12 and mebendazole lessens the severity of experimental eosinophilic meningitis caused by Angiostrongylus cantonensis in ICR mice.

Angiostrongylus cantonensis is the major cause of eosinophilic meningoencephalitis cases in Taiwan. Mice were orally infected with 35 infective larvae. One group of mice were given a single dose of mebendazole (20 mg/kg of body weight) per os at various times and examined at 14 days postinfection (dpi) for worm recovery rate and pathological studies. A 94 to 97% reduction in worm recovery was observed when medication was given at 4 to 5 dpi. Sections of the brains revealed that untreated infected mice developed typical severe eosinophilic meningoencephalitis. Meninges of these mice were thickened by massive infiltration of eosinophils, whereas only moderate pathological change was observed in the brains of mice that were treated with mebendazole at 4 dpi. Infected mice that received daily injections of 10 ng of interleukin-12 (IL-12) only for various numbers of days also exhibited moderate pathological changes in the brain. Eosinophil infiltration in the brains of these mice was low, and severe mechanical injuries in the parenchyma were observed. Treatment with mebendazole in combination with IL-12, however, resulted in low levels of worm recovery and dramatic lessening of the eosinophilic meningitis. A reverse transcriptase PCR assay of mRNA expression in the brain also revealed that the use of IL-12 had shifted the immune response of the mouse from Th2 type to Th1 type. This study could be used in developing strategies for the treatment of human angiostrongylosis.

Infectivity and pathogenicity of 14-month-cultured embryonated eggs of Toxocara canis in mice.

Infectivity and pathogenicity to mice of embryonated eggs of Toxocara canis, that had been maintained in 2% formalin for 14 months at 4 degrees C, were evaluated by immunological and pathological assessment at 1, 4, 8, 12, 16, 20, 24, 28, 42 and 67 weeks post-infection (WPI). On each date, three infected mice and two age-matched uninfected mice were sacrificed for serum collection and histological processing of the liver, lungs, musculature, and brain. Infectivity assessment by enzyme-linked immunosorbent assay (ELISA) revealed that the overall immunological pattern of infected mice tended to be towards the Th2 type response. Serum IgG1 antibody titers in infected mice were significantly higher than that of the uninfected control mice throughout the trial (P<0.05). On the other hand, no significant difference in titers of IgG3 antibody, an indicator for the Th1 type response, was observed between the infected and control mice, except at eight WPI (P<0.05). Pathogenicity was assessed semiquantitatively by comparing the mean number or diameter of inflammatory foci as well as histopathological changes in the liver, musculature, brain, or lungs of the infected mice and the control mice. Each hematoxylin and eosin (H&E) stained tissue section slide was examined under 100x magnification and 15 random fields were counted. Degree of inflammatory injury among the four organs was scored and categorized into four levels: normal (0), mild (1+), moderate (2+), and severe (3+). An index of inflammatory injury (mean score of experimental group/mean score of 10 control groups of 20 uninfected mice) of 2-3 is considered as moderate to severe, 1-2 as mild to moderate, and 0-1 as normal to mild. Histopathological changes were moderate to severe in the liver and lungs, mild to moderate in the musculature, and only normal to mild in the brain throughout the trial. It is noteworthy that apocrine-like change in epithelial cells of the bile duct was observed in most of the infected mice from eight WPI onward. Furthermore, larvae trapped by organized granulomas were found in soft tissue near the musculature at 12, 20, and 28 WPI. Altogether, not only were the infectivity and pathogenicity of the 14-month-cultured T. canis embryonated eggs retained, the hatched larvae were also capable of eliciting some special pathological changes in the murine host.