Validation of routine analytical method for injectable cyclosporine preparation control using HPLC-FIA assay.

OBJECTIVE: The aim of this study is to validate a new HPLC-FIA method for routine analytical control of cyclosporine injectable preparations and to evaluate the routine analytical control with this technic. MATERIAL AND METHODS: Cyclosporine dosage was carried out by the HPLC-FIA method. The column was replaced by a PEEK (polyetheretherketone) loop tubing. The mobile phase consisted of ultrapure water. The injection volume was 1µL with a flow rate of 1mL/min. All determinations were performed at 35°C. The detection was carried out at 210nm. The accuracy profile method was used to validate the HPLC-FIA assay of cyclosporine. Routine control was applied for each cyclosporine preparation using the HPLC-FIA developed method. An acceptance limit of ±10% of the theoretical concentration has been set for the conformity of the preparation. RESULTS: The accuracy profile shows the validity of our method for the dosage of cyclosporine in the concentration range studied (0.5-2.5mg/mL) with good linearity (correlation coefficient>0.999), high precision (the relative standard deviation [RSD] values, for both repeatability and intermediate precision, were<3%) and acceptable trueness (the relative biases were found<2%). In our study, 220 injectable cyclosporine preparations were analyzed: 85% were compliant. All analyzes were conform after a second standardized homogenization of 10 shakes. CONCLUSIONS: The proposed HPLC-FIA method is a reliable, fast, simple, precise method that can be easily used for the routine quality control of cyclosporine injectable preparations.

[Pseudotumoral sublingual leishmaniasis caused by Leishmaniainfantum]. / Leishmaniose sublinguale pseudotumorale à Leishmaniainfantum.

INTRODUCTION: Mucosal leishmaniasis is endemic in certain parts of Latin America and are usually absent in Morocco. Herein we report a case of Leishmaniainfantum in a Moroccan patient. PATIENTS AND METHODS: A 61-year-old male patient working as a tourist bus driver presented with a sublingual endobuccal tumor. He reported a history of treated cutaneous leishmaniasis of the lower lip in 2009 and had presented the sublingual oral tumor since December 2011. The histopathological findings as well as the species-specific PCR analysis confirmed the diagnosis of sublingual mucosal leishmaniasis due to L. infantum. HIV serology was negative. Our patient was then treated with intra-muscular meglumine antimoniate for 25 days, resulting in complete disappearance of the oral lesion. CONCLUSION: Our case thus has several peculiarities: the strictly mucosal character of the lesion, occurring in an immunocompetent subject, the unusual pseudotumoral form, and the causative agent, L. infantum, not known for its mucosal tropism in our country.

[Actinomycosic mycetoma of the foot in Morocco due to Actinomycetes viscosus]. / Mycétome actinomycosique du pied à Actinomycetes viscosus au Maroc.

We present the case of an actinomycotic mycetoma of the foot due to Actinomycetes viscosus. It evolved for nine years on the foot of a 26-year-old patient from a rural environment: Douar Inezgane (city in southern Morocco). Bacteriological study of the skin and grains confirmed the diagnosis. It showed positive bacilli on direct examination and on Gram staining and in positive culture. Histological study showed a polymorphous granulomatous inflammation without signs of malignancy with actinomycotic grains. Then we retained the diagnosis of primary cutaneous actinomycosis without visceral locations. The treatment was based on antibiotics: penicillin G by intravenous infusion for five weeks, relayed orally by amoxicillin associated with trimethoprim-sulfamethoxazole for long periods. After six months of treatment, we observed a favorable outcome with reduction of the swelling, nodules, lymphadenopathy, fistula's number and extension of time of issue of grains. The current follow up is 15 months. The primary cutaneous actinomycosis is still relevant in Morocco.

[Disseminated fusariosis during autologous stem cells transplant]. / Fusariose disséminée au cours d'une autogreffe de moelle osseuse.

Fusarium is a filamentous brown fungus found in soil, on plants and outdoors responsible for localized or disseminated infections. Diagnosis is based on blood cultures and skin biopsy. Disseminated fusariosis is a rare and serious fungal infection, that occurs especially in neutropenic immunosuppressed patients. Treatment is difficult and mortality is estimated between 50 and 70% in adult patients. This infection is rare in Morocco. We report a case of systemic fusariosis in patient with multiple myeloma during a second autologous stem cell transplant. At day 4 of the autologous stem cells transplant the patient had febrile neutropenia and diarrhea; he received ceftazidime, metronidazole and amikacin for 2 days. The patient still febrile was treated by imipenem and vancomycin without bacteriological proof. At day 10 the patient presented difficulty of breathing and wheezing on auscultation of the lungs, and received nebulization with salbutamol every 6 hours. The CT scan shows interstitial infiltrate of the right lung with micronodules. At day 11 he was treated by voriconazole with clinical improvement. At day 19, Fusarium sp. was identified on the Sabouraud blood culture. The patient left the transplant unit at day 25, he received 6 weeks of voriconazole with clinical and radiological improvement.

[Family violence]. / Violence familiale.

INTRODUCTION: Family violence is a serious public health problem, the scale of which is seriously increasing in Morocco. Although it has existed for a long time, we ignore the real characteristics of this plague in our country; our work consisted in an epidemiological approach of family violence in Marrakech during 2006. METHOD: After elaborating a questionnaire, which allows the study of the demographic and social profile of the families, the study of violence exercised in the family and the evaluation of the depression in the women, we led an inquiry amongst 265 women. RESULTS: Analysis of the results obtained has allowed us to underline the following characteristics: 16.6% of the women in our sample had been physically beaten; the young age is a risk factor; the age range most affected by violence is in women between the ages of 30 and 40 and which represent 39% of the battered women; domestic violence touches all the social, economic and cultural classes: in our study, 63% of the women having undergone violence were housewives, 25% were managers and 3% senior executives; family problems were the most important cause of violence in our study, representing 32.32%. Requests for money was the cause in 11.3% of the cases, and imposed sexual relations were found in 6.8% of the cases; alcoholism is an aggravating factor of family violence; 27.3% of the spouses who assaulted their wives were drunk; 52% of the assaulted women were victims of violence in childhood and 36% had been witness to their father's violence; in 63.6% of the cases of violence, the children were witnesses, and in 25% of the cases the children were victims of violence at the same time as their mothers; 50% of the women victims of violence did not react, while 38.6% left home, and 9.1 filed for divorce. Thirty-two percent of the assaulted woman had been traumatised by the aggression; the association of depression and violence was very high, 343% of the battered women in our study suffered from severe depression. CONCLUSION: This work underlines the necessity of an urgent intervention in order to limit the extension of this plague and its consequences.

[Generalized chromomycosis caused by Phialophora verrucosa]. / Chromomycose cutanée diffuse àPhialophora verrucosa.

BACKGROUND: Chromomycosis is a chronic fungal skin infection that generally presents in the form of verrucous or vegetative lesions on uncovered areas of skin. We report an unusual case of generalised chromomycosis due to Phialophora verrucosa. PATIENTS AND METHODS: A 42-year-old town-dwelling housewife was hospitalised for erythematous keratotic nodules on the arm showing sporotrichoid distribution, associated with a crusted ulcerative lesion on the homolateral index finger, as well as subcutaneous papulonodular lesions. In places, the lesions on the patient's back presented an umbilical and molluscoid appearance. The patient had suffered no previous injuries and had not visited any areas in which leishmaniasis is endemic. Her history included insulin-dependent diabetes. Screening for Leishman bodies was negative. Histopathological analysis of the skin biopsy revealed an epithelioid giant-cell granuloma with no caseous necrosis. The mycological study demonstrated the presence of fumagoid bodies and P. verrucosa was isolated. Treatment with terbinafine was initially given, followed by clarithromycin, but in the absence of any improvement, the patient was readmitted to hospital and is currently on itraconazole and amphotericin B. DISCUSSION: The novel features of our case comprise the clinical aspect of chromomycosis, the extent of the lesions, their unusual site on the back and upper limbs, and the isolation of a rare species, P. verrucosa (only the second observation in Morocco). It also highlights the therapeutic difficulties posed by this type of chromomycosis.

Central venous saturation is not an alternative to mixed venous saturation during cardiopulmonary bypass in coronary artery surgery patients.

BACKGROUND: To evaluate the correlation and agreement between central venous saturation (ScvO(2)) and mixed venous saturation (SvO(2)) during cardiopulmonary bypass. METHODS: Twenty-two consecutive patients scheduled for coronary artery surgery were prospectively included. Paired measurements of ScvO(2) and SvO(2) were performed 5 minutes after aortic cross-clamping, after each cardioplegia dose and after de-clamping of the aortic cross-clamp. ScvO(2) and SvO(2) were measured, respectively, by a fibreoptic catheter in the superior vena cava and on blood samples from the venous return line of the extracorporeal circuit, using a blood gas analyser RESULTS: Ninety-five paired measurements of venous saturation were obtained. Correlation between the measurements was associated with an r = 0.55. The mean bias was 2.2 [Limits of agreement: -13.6%, +18%]. Changes in oxygen saturation over time showed an r = 0.4 and a mean bias of 0.2 [Limits of agreement: -17.9%, +18.3%]. Multivariate analysis identified the oxygen consumption index as the only factor explaining this variability. CONCLUSIONS: Although mean biases between the measurements were low, limits of agreement were too large to provide a clinically acceptable estimation of SvO(2) by ScvO(2) in these conditions. Variations in regional oxygen consumption seem to be the main factor worsening the relationship.

Sandflies of the Phlebotomus perniciosus complex: mitochondrial introgression and a new sibling species of P. longicuspis in the Moroccan Rif.

The bloodsucking adult females of Phlebotomus perniciosus Newstead and P. longicuspis Nitzulescu (Diptera: Psychodidae) are important vectors of the protozoan Leishmania infantum Nicolle (Kinetoplastida: Trypanosomatidae) in western Mediterranean countries. The species status of the two phlebotomine sandflies was assessed, along with the epidemiological implications. Individual sandflies from three Moroccan Rif populations were characterized morphologically, isoenzymatically (by the isoelectrofocusing of alleles at the polymorphic enzyme loci of HK, GPI and PGM), and by comparative DNA sequence analysis of a fragment of mitochondrial Cytochrome b (mtDNA). By reference to the character profiles of specimens from other locations, including southern Spain and the type-locality countries, the Moroccan flies were placed in three lineages: first, the lineage of P. perniciosus, which contained two mtDNA sublineages, one (pnt) widely distributed and associated with the morphology of the male types from Malta, and the other (pna) associated with a P. longicuspis-like male morphology; second, the lineage of P. longicuspis sensu stricto, including typical forms from Tunisia; and third, a new sibling species of P. longicuspis. The mtDNA sublineage (pnt) of typical P. perniciosus was also found in some P. longicuspis from Morocco, indicating interspecific hybridization. The typical race of P. perniciosus occurs in Italy as well as in Malta, Tunisia and Morocco. It is replaced in southern Spain by the Iberian race (with the pni mtDNA sublineage). The discovery of interspecific gene introgression and a new sibling species mean that previous records of the two morphospecies do not necessarily reflect their true vectorial roles or geographical and ecological distributions.

[Canine leishmaniasis in the Rif mountains (Moroccan Mediterranean coast): a seroepidemiological survey]. / Leishmaniose canine dans le Rif (côte méditerranéenne marocaine): enquête séro-épidémiologique.

A sero-epidemiological survey has been conducted in several localities of the province of Nador to investigate canine leishmaniasis in the North-Eastern slope of the Rif mountains (Mediterranean coast of Morocco). Serum samples collected from 257 dogs were analysed using indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) to detect anti-Leishmania infantum antibodies. Forty eight (18.7%) of the screened dogs were IFAT positive and 54 (21.0%) were ELISA positive; the concordance of the two methods was 96.1%. The prevalence of infection is significantly higher in dogs more than four years of age whereas no significant difference in prevalence of infection was seen between males and females. The frequent symptoms observed in seropositive dogs were the enlargement of lymph nodes (57.4%), emaciation (51.9%) and skin involvement (25.9%). However, 38.9% of those dogs showed no one of the major symptoms of visceral leishmaniasis. Leishmania isolated from three of the examined dogs was identified as L. infantum MON-1. These results show that the North-Eastern slope of the Rif mountains is one of the most active Mediterranean areas of visceral leishmaniasis and confirm that the dog is the main reservoir of L. infantum.

An interdisciplinary research strategy to improve symbiotic nitrogen fixation and yield of common bean (Phaseolus vulgaris) in salinised areas of the Mediterranean basin.

The main findings of a cooperative research group of agronomists, plant breeders, microbiologists, physiologists and molecularists to improve the symbiotic nitrogen fixation (SNF) and N2-dependent yield of common bean under moderate salinity in the Mediterranean basin are summarised. Agronomic surveys in reference production areas show large spatial and temporal variations in plant nodulation and growth, and in efficiency of utilisation of the rhizobial symbiosis. The latter was associated with a large rhizobial diversity, including new bean nodulating species. Macrosymbiont diversity in SNF and adaptation to NaCl was found. However, contrasts between plant genotypes could be altered by specific interactions with some native rhizobia. Therefore, variations in soil rhizobial population, in addition to agronomic practices and environmental constraints, may have contributed to erratic results observed in field inoculations. At the mechanistic level, nodule C and N metabolisms, and abcissic acid content, were related to SNF potential and tolerance to NaCl. Their relation with nodule conductance to O2 diffusion was addressed by in situ hybridisation of candidate carbonic anhydrase and aquaporin genes in nodule cortex. The limits and prospects of the cooperative strategy are discussed.

Effects of salinity on protein and lipopolysaccharide pattern in a salt-tolerant strain of Mesorhizobium ciceri.

AIMS: To characterize the physiological and metabolic responses of Mesorhizobium ciceri strain ch-191 to salt stress, investigating the changes induced by salinity in protein and lipopolysaccharide profiles, as well as determining the accumulation of amino acids, glutamate and proline. METHODS AND RESULTS: Strain ch-191 of M. ciceri was grown with different NaCl concentrations. Protein and lipopolysaccharide patterns were determined by electrophoresis. The strain ch-191 tolerated up to 200 mmol l-1 NaCl, although higher salt dosages limited its growth and induced changes in the protein profile. The most noteworthy change in the LPS-I pattern was the decrease in the slowest band and the appearance of an intermediate mobility band. The accumulation of proline in response to salt stress surpassed that of glutamate. CONCLUSION: The protein profile showed major alterations at salinity levels which inhibited growth. However, the alterations in the LPS profile and accumulation of compatible solutes were evident from the lowest levels, suggesting that these changes may constitute adaptative responses to salt, allowing normal growth. SIGNIFICANCE AND IMPACT OF THE STUDY: The selection and characterization of salt-tolerant strains, which also show efficient symbiotic performance under salinity, may constitute a strategy for improving Cicer arietinum-Mesorhizobium ciceri symbiosis in adverse environments.

Morphological and isoenzymatic differentiation of sympatric populations of Phlebotomus perniciosus and Phlebotomus longicuspis (Diptera: Psychodidae) in northern Morocco.

The phlebotomine sandfly Phlebotomus perniciosus Newstead is a vector of Leishmania infantum Nicolle in the western Mediterranean basin. In northern Morocco it is often caught in sympatry with a closely related species, P. longicuspis Nitzulescu. In the locality of Chefchaouene where females of both species were morphologically distinguishable, none of the collected males exhibited forked copulatory valves, which is considered a specific character for P. perniciosus. They 1st were identified as P. longicuspis. Isoelectrofocusing of 7 enzyme systems was used to compare these specimens with the sympatric females. Two groups of males were separated and assigned to each species by their isoenzyme patterns associated with morphological differences in the shape of the aedeagus and in the number of coxite hairs. Based on our results, a reexamination of the distribution of both species is necessary especially in many locations where P. longicuspis has been identified only from males.