Murine neonatal infection provides an efficient model for congenital ocular toxoplasmosis

Congenital infection is one of the most serious settings of infection with the apicomplexan parasite Toxoplasma gondii. Ocular diseases, such as retinochoroiditis, are the most common sequels of such infection in utero. However, while numerous studies have investigated the physiopathology of acquired toxoplasmosis, congenital infection has been largely neglected so far. Here, we establish a mouse model of congenital ocular toxoplasmosis. Parasite load and ocular pathology have been followed for the first 4 weeks of life. Ocular infection developed slowly compared to cerebral infection. Even after 4 weeks, not all eyes were infected and ocular parasite load was low. Therefore, we evaluated a scheme of neonatal infection to overcome problems associated with congenital infection. Development of infection and physiopathology was similar, but at a higher, more reliable rate. In summary, we have established a valuable model of neonatal ocular toxoplasmosis, which facilitates the research of the underlying physiopathological mechanisms and new diagnostic approaches of this pathology.     

Refinement of the mouse model of congenital toxoplasmosis

The goals of the present investigation, focusing on the BALB/c mouse model of congenital toxoplasmosis, were: (1) to find a method to determine pregnancy in the mouse. The method has 100% sensitivity and 72% specificity; (2) to test congenital transmission during the chronic stage of toxoplasmosis. This occurred in 2 of 10 mice tested; (3) to investigate the relationship between the infective dose and the rate of congenital transmission. This was not demonstrated for doses of 10(2) to 10(3) bradyzoites and oocysts of Prugniaud, M3 and M7741 strains, with transmission rates of 3 of 8 to 6 of 10 mice inoculated; (4) to determine homologous and heterologous protection. Homologous protection was demonstrated with Prugniaud cysts, and heterologous protection was found between ME-49 and M3 cysts. This finding is consistent with the uniform natural protection against congenital toxoplasmosis seen in immune women and ewes.     

Clinical features of ocular toxoplasmosis in Korean patients

We report here the records of 10 consecutive Korean patients (10 eyes) with ocular toxoplasmosis which showed the typical clinical manifestations with seropositivity for Toxoplasma gondii specific IgG antibodies by micro-ELISA between 2006 and 2010. Nine patients were males and 1 was female; their age was 50.5 ± 13.8 years. The most common accompanying signs were vitritis (100%), anterior uveitis (70%), and scattered white deposit (80%). Pre-existing retinochoroidal scar was found in 1 (10%) patient. All patients received antiparasitic chemotherapy and systemic corticosteroid treatment, which resolved the presenting attack and recovered the visual acuity better than initial one in 9 patients and worse in 1. Optic atrophy, cataract, and retinal neovascularization were observed during the follow-up period and recurrence was detected in 3 eyes (30%) 6 to 20 months after the initial attack. In Korea, although rarely detected and reported, ocular toxoplasmosis needs more attention in clinical field of retinal diseases.     

New clinical and experimental insights into Old World and neotropical ocular toxoplasmosis

Retinal lesions or other ocular manifestations are serious consequences of infection with the protozoan parasite Toxoplasma gondii. Whilst classically considered a consequence of congenital transmission, recent screening studies estimated that 2% of T. gondii seropositive persons in Europe and North America have retinal lesions, most of them persisting unnoticed. The situation is more dramatic in South America, probably due to the predominance of virulent strains. Some of these strains seem to exhibit ocular or neuronal tropism and are responsible for severe ocular lesions. Despite the medical importance, the physiopathological mechanisms have only recently begun to be elucidated. The particular immune-privileged situation in the eye has to be considered. Studies on French patients showed low or undetectable ocular parasite loads, but a clear Th1/Th17 type immune reaction. Suitable mouse models have appeared in the last few years. Using such a model, IL-17A proved to impair parasite control and induce pathology. In contrast, in South American patients, the parasite seems to be much less efficiently controlled through a Th2 type or suppressive immune response that favors parasite replication. Finally, several host genetic markers controlling immune response factors have been associated with ocular involvement of T. gondii infection, mainly in South America.     

Toxoplasma gondii: partial cross-protection among several strains of the parasite against congenital transmission in a rat model

Rats were immunized with cysts of two Toxoplasma strains or with RH strain tachyzoites prior to pregnancy. The litters of the 13 rats that received homologous challenges with cysts during pregnancy, were all protected, whereas of 173 rats that received heterologous challenges with cysts or oocysts, only 21 protected their litters. 38.3 and 17% of rats immunized with the RH and with complete strains respectively, and 57% of control rats challenged with cysts, transmitted the infection congenitally. The percentages when similar groups were challenged with oocysts, were 33.3, 48.2, and 56.2%, respectively. Immunization with cysts did not completely protect against challenge with oocysts, even if the same strain was used. The divergence of these results from the complete protection against congenital toxoplasmosis observed in immune women and ewes, might be due to the use of excessive challenge doses in the model.     

脑室—腹腔分流术后感染26例诊治分析

目的探讨脑室—腹腔分流术后感染的早期诊断和治疗原则。方法回顾性分析26例脑室—腹腔分流术后感染病例的诊断和治疗办法,所有病例均进行血液和脑脊液的培养及影像学检查。结果 2例死于感染并发症,2例感染复发,21例患者重新安装分流装置,3例病情较轻的患者经分流泵及鞘内给药后痊愈。结论脑室—腹腔分流术后的早期诊断非常重要,一些非特异性的症状和体征需引起重视。对于感染的病例,及时移除分流装置,同时行脑室外引流冲洗和脑室内注药,当脑脊液达到无菌状态后重新植入分流装置是行之有效的办法。结果表明抗感染应首选万古霉素、第三代头孢菌素或改为美罗培南,待获得培养结果后根据药敏试验针对性用药。脑脊液培养以金黄色葡萄球菌和表皮葡萄球菌多见,经治疗再次培养结果均为阴性。     

An overview on the correlation of neurological disorders with cardiovascular disease

Neurological disorders (NDs) are one of the leading causes of death especially in the developed countries. Among those NDs, Alzheimer’s disease (AD) and Parkinson disease (PD) are heading the table. There have been several reports in the scientific literatures which suggest the linkage between cardiovascular disorders (CVDs) and NDs. In the present communication, we have tried to compile NDs (AD and PD) association with CVDs reported in the literature. Based on the available scientific literature, we believe that further comprehensive study needs to be done to elucidate the molecular linking points associated with the above mentioned disorders.Abbreviations: AD, Alzheimer’s disease, Aβ, β amyloid, PD, Parkinson disease, l-DOPA, l-dihydroxyphenylalanine, LBs, Lewy bodies, DA, dopamine, APP, amyloid precursor protein, CVD, cardiovascular disease     

布氏杆菌脑膜炎病例报道2例及文献复习

目的:目前布氏杆菌性脑膜炎在国内只是偶见报道,本文报道2例布氏杆菌性脑膜炎,对其诊断及治疗进行探讨,并对布氏杆菌脑膜炎进行文献回顾.方法:我们近期连续通过检查脑脊液内布氏杆菌抗体的办法诊断了2例布氏杆菌性脑膜炎,并通过给予四环素、利福霉素及链霉素治疗1个月并通过随访.结果:半年后脑脊液内布氏杆菌抗体恢复阴性,临床症状完全消失.结论:通过我们的观察应用上述3联药物综合治疗1月对布氏杆菌性脑膜炎是有效的.     

Fitness of Toxoplasma gondii is not related to DHFR single-nucleotide polymorphism during congenital toxoplasmosis

Factors that regulate the pathogenesis of Toxoplasma gondii in humans are poorly understood. When acquired during pregnancy, toxoplasmosis can be disastrous, leading to fetal loss or conversely to subclinical disease. In congenitally infected infants, evolution is highly unpredictable. Genotype based virulence patterns have been described in mice, but in humans this classification does not correlate with the gravity of the disease. Mutations on DHFR-TS loci have recently been reported to confer T. gondii fitness cost. In this study, we investigated the relationship between the virulence of the parasite, as measured by clinical outcome in the fetus or newborn, fitness, as measured by parasitic load in amniotic fluid, and allelic polymorphism in DHFR. Six cases of severe congenital toxoplasmosis and 23 cases of mild congenital infections were included in the study. Quantitative PCR was performed to evaluate total T. gondii DNA load in amniotic fluid and detection of mutations was carried out with a LightCycler using hybridisation probes. Parasitic load was significantly higher in severe infections than in mild diseases. Among isolates from severe or non-severe cases of congenital toxoplasmosis, no polymorphism could be detected at loci 36, 83 or 245 of the DHFR gene. The virulent RH strain presented the same melting temperature as the non-virulent PRU strain for codons 36, 83 and 245. Only mutated clones, M2M3 and M2M4 with allelic replacement at these positions, displayed different profiles allowing a clear distinction between wild and mutant types. We concluded that the DHFR gene mutations we investigated do not regulate T. gondii fitness in humans.     

Evaluation of a recombinant rhoptry protein 2 enzyme-linked immunoassay for the diagnosis of toxoplasmosis acquired during pregnancy

The aim of this study was to evaluate an enzyme-linked immunoassay with recombinant rhoptry protein 2 (ELISA-rROP2) for its ability to detectToxoplasma gondii ROP2-specific IgG in samples from pregnant women. The study included 236 samples that were divided into groups according to serological screening profiles for toxoplasmosis: unexposed (n = 65), probable acute infection (n = 48), possible acute infection (n = 58) and exposed to the parasite (n = 65). When an indirect immunofluorescence assay forT. gondii-specific IgG was considered as a reference test, the ELISA-rROP2 had a sensitivity of 61.8%, specificity of 62.8%, predictive positive value of 76.6% and predictive negative value of 45.4% (p = 0.0002). The ELISA-rROP2 reacted with 62.5% of the samples from pregnant women with probable acute infection and 40% of the samples from pregnant women with previous exposure (p = 0.0180). Seropositivity was observed in 50/57 (87.7%) pregnant women with possible infection. The results underscored that T. gondii rROP2 is recognised by specific IgG antibodies in both the acute and chronic phases of toxoplasmosis acquired during pregnancy. However, the sensitivity of the ELISA-rROP2 was higher in the pregnant women with probable and possible acute infections and IgM reactivity.     

Effect of zinc and melatonin supplementation on cellular immunity in rats with toxoplasmosis

The effects of zinc (Zn) and/or melatonin supplementation on cellular immunity were investigated in rats infested with Toxoplasma gondii. Fifty Sprague-Dawley male rats were used for this study. All animals were fed a normal diet, ad libitum, containing 97 mg Zn/kg. They were divided into five experimental groups, as follows. Group I (n=10) received intraperitoneal injections of zinc sulfate at a dose of 3 mg/kg/d for 3 wk. Group II (n=10) received intraperitoneal injections of melatonin at a dose of 3 mg/kg/d for 3 wk. Group III (n=10) received intraperitoneal injections of zinc sulfate (3 mg/kg/d) and melatonin (3 mg/kg/d) for 3 wk. Group IV (n=10) was infested controls. Group V (n=10) was healthy controls. There were no differences in the percentage of CD3+ lymphocytes among all groups. For groups I–III, the CD4+ and CD8+ ratios were higher than those of the groups IV and V controls (p<0.01). Similarly, the total lymphocyte ratios in groups I–III were higher than those of infested and healthy controls (p<0.01). The total lymphocyte ratios in group III were significantly higher than those of groups I and II (p<0.01). The plasma Zn levels in the supplemented groups were significantly higher than those of control groups IV and V (p<0.01). These results suggest that melatonin and/or Zn supplementation may activate cellular immunity by stimulating CD4+ and CD8+ production in infected rats with T. gondii.     

先天性唇腭裂病400例病因探讨

目的：探讨先天性唇腭裂病的病因,寻找预防办法。方法：对先天性400例唇腭裂手术患者的父母在怀孕前后居住生活、饮食卫生、嗜好、心情精神状态及患病情况做统计分析。结果：400例患者父母怀孕前后居住面积在30m^2以下占75％,饮酒吸烟者人数627人中78．3％,女性饮酒167人占41．07％。患普通感冒730人。怀孕在1月内患感冒283人,心情不愉快精神毋张受到不良刺激362人。结论：通过对400例患者父母怀孕前后生活居住患病嗜好精神等因素情况得出结论与外界环境关系密切,尤其怀孕早期患病饮酒吸烟心情不愉快对该病发生起一定作用,但该病是多种原因综合因素所致的一种先天性疾病,具有一定遗传性,但也有可以预防的一方面,在谊病发生的过程中有一个主要因素起主导作用,有待今后继续研讨。     

Toxoplasma gondii: genotyping of strains from Brazilian AIDS patients with cerebral toxoplasmosis by multilocus PCR-RFLP markers

This study investigated the genetic characteristics of the Toxoplasma gondii strains isolated from 87 patients with cerebral toxoplasmosis and AIDS, treated in Sao Paulo State, Brazil. The laboratorial diagnosis of cerebral toxoplasmosis was based on positive serological exams and PCR of blood and/or cerebrospinal fluid. Four markers (5'-SAG2, 3'-SAG2, SAG3 and GRA6) were chosen to analyze the samples. Each having clear resolution to distinguish the three clonal lineages after PCR amplified targets were treated with restriction enzyme digestion (PCR-RFLP). The genotyping provided the following results: 40 patients (46%) were infected with strains classified as type I; 4 (4%), as type III; 13 (15%) were infected with polymorphic strains (unusual genotype); 6 patients with type I or II alleles; and 15 (17%) patients had strains not classified for any marker. PCR-RFLP, also classified 9 (11%) clinical isolates as type II, which is uncommon in South America. However, the sequencing of the nested-PCR products (of SAG3 marker) of type II and polymorphic isolates (of 5'-SAG2, SAG3 and GRA6 markers) showed a nucleotide polymorphism compared with the archetypal clonal genotypes (types I, II and III) and these isolates were considered as polymorphic strains. The markers used here were inappropriate to distinguish the most isolates considered as polymorphic strains. These data confirm other studies showing the high rate of genetic polymorphism in T. gondii strains isolated in Brazil.     

先天性唇腭裂病400例病因探讨

目的:探讨先天性唇腭裂病的病因,寻找预防办法。方法:对先天性400例唇腭裂手术患者的父母在怀孕前后居住生活、饮食卫生、嗜好、心情精神状态及患病情况做统计分析。结果:400例患者父母怀孕前后居住面积在30m2以下占75%,饮酒吸烟者人数627人中78.3%,女性饮酒167人占41.07%。患普通感冒730人。怀孕在1月内患感冒283人,心情不愉快精神紧张受到不良刺激362人。结论:通过对400例患者父母怀孕前后生活居住患病嗜好精神等因素情况得出结论与外界环境关系密切,尤其怀孕早期患病饮酒吸烟心情不愉快对该病发生起一定作用,但该病是多种原因综合因素所致的一种先天性疾病,具有一定遗传性,但也有可以预防的一方面,在该病发生的过程中有一个主要因素起主导作用,有待今后继续研讨。     

外科急腹症48例临床诊疗分析

目的:探讨外科急腹症的早期诊断与治疗,以便总结经验,提高治愈率,降低致残率与病死率。方法:回顾性分析48例外科急腹症诊治的临床资料。结果:急诊手术40例,经严密观察后手术8例,术后并发症11例,治愈44例,死亡4例。结论:外科急腹症的诊断应结合病史,体征及辅助检查等综合判断。对其早期确诊与及时处置和手术治疗是降低病死率的关键。