High-Throughput Sequencing for Trypanosome Transcriptome Characterization.

High-throughput sequencing of cDNA (RNASeq) is now the method of choice for analysis of transcriptomes. This chapter details important considerations in the design of RNASeq experiments for kinetoplastids grown in culture or experimental animals. It contains protocols for obtaining parasites from rodents, and for removal of rRNA from total RNA. In addition, custom pipelines for sequence alignment, and for data analysis and visualization, are described.

Cerebral trypanosomiasis and AIDS.

UNLABELLED: A 36 year-old black female, complaining of headache of one month's duration presented with nausea, vomiting, somnolence, short memory problems, loss of weight, and no fever history. Smoker, intravenous drugs abuser, promiscuous lifestyle. PHYSICAL EXAMINATION: left homonimous hemianopsia, left hemiparesis, no papilledema, diffuse hyperreflexia, slowness of movements. Brain CT scan: tumor-like lesion in the splenium of the corpus calosum, measuring 3.5 x 1.4 cm, with heterogeneous enhancing pattern, suggesting a primary CNS tumor. Due to the possibility of CNS infection, a lumbar puncture disclosed an opening pressure of 380 mmH(2)0; 11 white cells (lymphocytes); glucose 18 mg/dl (serum glucose 73 mg/dl); proteins 139 mg/dl; presence of Trypanosoma parasites. Serum Elisa-HIV tests turned out to be positive. Treatment with benznidazole dramatically improved clinical and radiographic picture, but the patient died 6 weeks later because of respiratory failure. T. cruzi infection of the CNS is a rare disease, but we have an increasing number of cases in HIV immunocompromised patients. Diagnosis by direct observation of CSF is uncommon, and most of the cases are diagnosed by pathological examination. It is a highly lethal disease, even when properly diagnosed and treated. This article intends to include cerebral trypanosomiasis in the differential diagnosis of intracranial space-occupying lesions, especially in immunocompromised patients from endemic regions.

Combination of eflornithine and melarsoprol for melarsoprol-resistant Gambian trypanosomiasis.

OBJECTIVE: To evaluate the efficacy and toxicity of a combination of eflornithine and melarsoprol among relapsing cases of Gambian trypanosomiasis. METHODS: Forty-two late-stage Trypanosoma brucei gambiense trypanosomiasis patients relapsing after initial treatment with melarsoprol were treated with a sequential combination of intravenous eflornithine (100 mg/kg every 6 h for 4 days) followed by three daily injections of melarsoprol (3.6 mg/kg, up to 180 mg). They were then followed-up for 24 months. RESULTS: Two (4.8%) patients died during treatment. Of the 40 surviving patients, two had a treatment failure, 13 and 19 months after having received the combination therapy. By Kaplan-Meier analysis, the 2-year probability of cure was 93.3% (95% confidence interval: 84.3-100%). CONCLUSION: This sequential combination has an efficacy and a toxicity similar to a 7-day course of eflornithine monotherapy, but is easier to administer. Whether such therapeutic success corresponds tosynergism between eflornithine and melarsoprol, or merely means that 4 days of eflornithine monotherapy suffices for such patients, will need to be determined in a comparative trial.

Detection of Trypanosoma evansi in brains of the naturally infected hog deer by streptavidine-biotin immunohistochemistry.

Twenty-four percent of hog deer (Cervus porcinus) that ranged free on a farm in Samut Prakarn province, Thailand, died showing nervous signs between September 1997 and February 1998. The nervous signs shown by most of them included ataxis, paresis of hind limbs, lateral recumbency, excitation and convulsion. Six animals and one carcass were submitted for diagnosis at the National Institute of Animal Health, Bangkok. Trypanosoma evansi was detected in blood and cerebrospinal fluid of four and five animals, respectively. Antibodies to T. evansi were found in all the hog deer by indirect enzyme-linked immunosorbent assay. Histopathological observation revealed a generalised non-suppurative meningoencephalitis affecting the white and grey matter at all levels of the brain. Typically, there were broad perivascular cuffs of mononuclear inflammatory cells, including lymphocytes, and some Mott cells. No trypanosomes were found in any tissue examined by conventional histopathology. However, numerous T. evansi were demonstrated by streptavidine-biotin immunohistochemistry in neuropil and Virchow-Robin spaces of brain in three animals.

[Indirect immunofluorescence in the therapeutic surveillance of trypanosomiasis (preliminary note)]. / L'immunofluorescence indirecte dans la surveillance thérapeutique des trypanosomes (note préliminaire).

The authors have studied the evolution of fluorescent antibodies in the serum and the C. S. F. of treated Sleeping Sickness patients regularly examined when coming to Brazzaville for their periodical controls. They have observed more than 200 patients who were followed during three years. The authors demonstrated that curing patients show the following particularities: -- serum becomes negative less rapidly than C. S. F., -- delay of negativation of the serum varies according to the stage of the disease, -- the C. S. F. positive in fluorescence becomes negative in less than twelve months. On the other hand, relapsing patients show some striking differences, such as: -- C. S. F. remains always strongly fluorescent positive, -- the serology remains positive too, but can present some fluctuations. Some very particular cases such as "nuclear fluorescence only" or discrepancy between IFAT and clinico-parasitological data are then analysed. From a practical point of view, it appears that, as often as not, the evolution of fluorescent antibodies in serum and in C. S. F. allows to prognose either the cure or the relapse less than one year after treatment.