Leishmania major cutaneous leishmaniasis outbreak in the Jordanian side of the Northern Jordan Valley.

Cutaneous leishmaniasis (CL) is endemic in many foci of Jordan and the Jordanian Mid Jordan Valley (JMJV) is the most affected and the incidence is quite high. The situation in the northern part of the Jordanian side of the Jordan Valley (NJJV) was different; before 2008, CL has rarely been reported from this area. From April 2008 to May 2009, passive detection followed by active detection was used to trace cases of CL from the NJJV. DNA was extracted from seven clinical isolates of Leishmania promastigotes and lesion scrapings spotted on filter papers obtained from 51 suspected CL patients living in the NJJV. The identity of the causative species of CL in the NJJV was investigated using ITS1-PCR followed by RFLP. In 2008/2009, 183 cases were clinically diagnosed of having CL in the NJJV. The parasites in five of the isolates and in 48 PCR-positive scrapings were classified as Leishmania major. In two isolates and in one PCR-positive scraping Leishmania tropica was identified. Investigations on the origin of CL cases revealed that the L. tropica cases were residents of two towns outside the NJJV. Herein, we report the clinical features, parasitological diagnosis, etiology, and the geographical distribution of CL cases from NJJV with the aim of documenting, for the first time, an outbreak in this area.

The Jordanian Mid Jordan Valley is a classic focus of Leishmania major as revealed by RFLP of 56 isolates and 173 ITS-1-PCR-positive clinical samples.

The identity of the causative species of cutaneous leishmaniasis (CL) in the endemic Jordanian Mid Jordan Valley (JMidJV) was investigated using the polymerase chain reaction (PCR) amplifying the ribosomal internal transcribed spacer 1 (ITS-1) followed by the restriction fragment length polymorphism (RFLP). The geographical distribution of CL and the usefulness of ITS1 PCR in diagnosis of suspected CL in the study area were also addressed. Over the period from 2004 to 2009, 56 clinical isolates of Leishmania promastigotes and 185 lesion scrapings spotted on filter papers were obtained from suspected CL patients living in the JMidJV, which is divided into northern and southern districts. The majority (67.1%) of patients occurred in the populated eastern part of the southern district. Of the 185 suspected CL patients, 173 (93.5%) were confirmed positive using PCR. Leishmanial DNA was detected in 27 (90%) of 30 patients having clinically atypical lesions of CL and in 60 (92%) of 65 smear- and culture-negative cases having typical lesions of CL. The parasites in all of the 56 isolates and the 173 PCR-positive scrapings were classified as Leishmania major. In conclusion, PCR is useful in diagnosis of CL especially when smear and culture are negative. It is also recommended as a differential diagnostic tool of atypical lesions when CL is endemic. The identification of L. major as the causative species in such a considerable number of CL cases, representative of all mini foci of CL in the study area, shows that the JMidJV is a classic focus of L. major.

Chemical constituents of Osyris alba and their antiparasitic activities.

Phytochemical investigation of Osyris alba L. (Santalaceae) of Jordanian origin resulted in the isolation and identification of one new pyrrolizidine alkaloid, osyrisine (1), together with 16 other known compounds. The structures of all compounds were established on the basis of spectroscopic analysis. Osyrisine, catechin, and catechin-3-O-α-L-rhamnopyranoside exhibited a significant level of antiparasitic activity against two parasites, Entamoeba histolytica and Giardia intestinalis.

New synthesis and antiparasitic activity of model 5-aryl-1-methyl-4-nitroimidazoles.

A number of 5-aryl-1-methyl-4-nitroimidazoles 5a-f have been synthesized in good yields by the Suzuki coupling reaction between 5-chloro-1-methyl-4-nitroimidazole (3) and arylboronic acids 4a-f, aided by dichlorobis-(triphenylphosphine)palladium(II), K(2)CO(3, )and tetrabutylammonium bromide in water at 70-80 degrees C. Compounds 5a-f were characterized by elemental analysis, NMR and MS spectral data. On the basis of in vitro screening data, 5-(3-chlorophenyl)-1-methyl-4-nitro-1H-imidazole (5f) exhibited potent lethal activity against Entamoeba histolytica and Giardia intestinalis with IC(50) = 1.47 microM/mL, a value lower by a factor of two than that of the standard drug, metronidazole. The boosted activity of 5f was not accompanied by any increased cytotoxicity.The rest of the series also exhibited potent antiparasitic activity with IC(50 ) values in the 1.72-4.43 microM/mL range. The cytotoxicity of the derivatives 5c and 5e was increased compared to the precursor compound, metronidazole, although they remain non-cytotoxic at concentrations much higher than the antiparasitic concentration of the two derivatives.

Synthesis of novel hybrid molecules from precursors with known antiparasitic activity.

Three novel new compounds derived from antiparasitic precursors have been synthesized and tested for their antiamoebic and antigiardial activities. The condensation of 2-(2-methyl-5-1H-nitroimidazolyl)ethylamine (6) with 5-nitro-2-furylacrylic acid (7) gave 3-(5-nitrofuran-2-yl)-N-[2-(5-nitroimidazol-1-yl)ethyl]acrylamide (8). Condensation of 7 with 7-chloro-4-(piperazin-1-yl)quinoline (9) afforded 1-[4-(7-chloroquinolin-4-yl)piperazin-1-yl)-3-(5-nitrofuran-2-yl)propenone as a mixture of two isomers; 10-a (the E-isomer) and 10-b (the Z-isomer). In addition, the reaction of 9 with 1-(2-bromoethyl)-2-methyl-5-nitroimidazole (11) in the presence of K(2)CO(3) and NaI yielded 7-chloro-4-(4-[2-(5-nitroimidazol-1-yl)ethyl]-piprazin-1-yl)quinoline (12). On the basis of preliminary screening data for these new compounds, compound 12 exhibited potent lethal activities against Entamoeba histolytica and Giardia intestinalis; its IC(50) (about 1 microM) was lower, at least by a factor of five, compared to the standard drug, metronidazole. In addition, the IC(50) of compound 12 against the tested parasites is 600 times below that against Hep-2 and Vero cells. Compounds 8 and 10-a also exhibited potent or moderate antiamoebic and antigiardial activities with IC(50 values) of about 5.5 microM, and 140 microM, respectively, against the tested parasites. These two hybrid molecules, 8, 10-a, were also non-cytotoxic at the lethal concentrations against the parasites.

Cutaneous leishmaniasis in the Jordanian side of the Jordan Valley: severe under-reporting and consequences on public health management.

OBJECTIVE: To test if cutaneous leishmaniasis (CL) is under-reported in the Jordanian Mid Jordan Valley, with resultant serious consequences for drug supply. METHODS: For 2001-2003, prescribed amounts of drug and laboratory log-books were investigated to estimate CL cases reported in Jordanian Mid Jordan Valley. From April 2004 to May 2005, passive detection and focused active 'index case cluster'-directed detection were used. RESULTS: An average of 75/100,000 cases per year was estimated to have occurred 2001-2003, resulting in under-reporting by a factor of 47. In 2004/2005, 313/100,000 cases per year were passively detected. Active case-finding detected additional cases. CONCLUSION: Cutaneous leishmaniasis is severely under-reported in Jordanian Mid Jordan Valley, which impacts its eradication.

Efficacy of a weekly cryotherapy regimen to treat Leishmania major cutaneous leishmaniasis.

BACKGROUND: Antileishmanial chemotherapy can have adverse effects and may fail to cure patients with cutaneous leishmaniasis (CL). OBJECTIVE: To evaluate the efficacy of a weekly cryotherapy regimen in patients with confirmed Leishmania major infection. METHODS: One hundred twenty Jordanian patients with 375 lesions were treated with cryotherapy performed once weekly in 1 to 7 sessions. This regimen was adopted since living parasites were recovered, in several CL patients, even after 3 cryosessions. Scarring was assessed in 78 patients 3 years after treatment. RESULTS: Approximately 84% of the lesions were cured after 1 to 4 sessions. The remaining lesions (16.3%) were cured after an additional 1 to 3 session(s). Cryotherapy caused mild adverse side effects and most of the patients were cured with negligible scarring. Statistically, the lesion size and location significantly affected the clinical response to cryotherapy. LIMITATIONS: A fraction of patients was followed up 3 years after healing. CONCLUSION: Cryotherapy with weekly intervals for 1 to 4 sessions is effective to treat L major CL, especially for smaller lesions. Extra sessions may be necessary to cure larger lesions. The cosmetic results are very good and no relapses were registered.