Evaluation of the Antiplasmodial Activity and Lethality of the Leaf Extract of Cassia alata L. (Fabaceae).

OBJECTIVE: Cassia alata L. (Fabaceae), one of the three plants contained in Saye, a polyherbal antimalarial remedy was assessed for its antimalarial potential and safety in mice. METHODOLOGY: Organic extracts were prepared from the leaves and tested on the D 10 chloroquine-sensitive strain of Plasmodium falciparum using the parasite lactate dehydrogenase assay. The 4 days suppressive test using Plasmodium berghei in mice was used to evaluate the in vivo antiplasmodial activity of the extracts. Animals were treated by oral route, once a day with 50, 100, 250 and 400 mg kg -1 b.wt., of the extracts. The acute toxicity of the extracts was assessed in mice according to Thompson and Weil method. The lethal effects of the extracts on animal's body weight, tissues, biochemical and haematological parameters were determined at 823.5, 1235.5, 1853 and 2779.5 mg kg -1 b.wt., respectively. RESULTS: The dichloromethane/methane (1:1, v/v) extract of Cassia alata was the most active against Plasmodium falciparum. The mean percent suppression of parasitemia in mice was equal to 22.5, 41.8 and 45.2% at 50, 250 and 400 mg kg -1 b.wt., respectively. No death and no clinically significant changes were recorded in mice. The maximum non-lethal dose was more than 16875 mg kg -1 in animals. No significant changes were observed in body weight, tissues morphology, biochemical and hematological parameters at doses above or equal to 2779.5 mg kg -1 b.wt. CONCLUSION: The dichloromethane/methanol leaf extract of Cassia alata had a good to moderate in vitro and in vivo antiplasmodial activity and was found to have low toxicity at high doses in tested animals.

Combining rapid diagnostic tests and dried blood spot assays for point-of-care testing of human immunodeficiency virus, hepatitis B and hepatitis C infections in Burkina Faso, West Africa.

People screened for human immunodeficiency virus (HIV) using rapid diagnostic tests (RDTs) in Africa remain generally unaware of their status for hepatitis B (HBV) and hepatitis C (HCV) infections. We evaluated a two-step screening strategy in Burkina Faso, using both HIV RDTs and Dried Blood Spot (DBS) assays to confirm an HIV-positive test, and to test for HBV and HCV infections. HIV counselling and point-of-care testing were performed at a voluntary counselling and testing centre with HBV, HCV status and HIV confirmation using DBS specimens, being assessed at a central laboratory. Serological testing on plasma was used as the reference standard assay to control for the performance of DBS assays. Nineteen out of 218 participants included in the study were positive for HIV using RDTs. A fourth-generation HIV ELISA and immunoblot assays on DBS confirmed HIV status. Twenty-four out of 25 participants infected with HBV were found positive for hepatitis B surface antigen (HBsAg) using DBS. One sample with a low HBsAg concentration on plasma was not detected on DBS. Five participants tested positive for HCV antibodies were confirmed positive with an immunoblot assay using DBS specimens. Laboratory results were communicated within 7 days to participants with no loss to follow up of participants between the first and second post-test counselling sessions. In conclusion, DBS collection during HIV point-of-care testing enables screening and confirmation of HBV, HCV and HIV infections. Diagnosis using DBS may assist with implementation of national programmes for HBV, HCV and HIV screening and clinical care in middle- to low-income countries.

Population pharmacokinetics and pharmacodynamics of piperaquine in children with uncomplicated falciparum malaria.

Dihydroartemisinin-piperaquine is being increasingly used as a first-line artemisinin combination treatment for malaria. The aim of this study was to describe the pharmacokinetic and pharmacodynamic properties of piperaquine in 236 children with uncomplicated falciparum malaria in Burkina Faso. They received a standard body weight-based oral 3-day fixed-dose dihydroartemisinin-piperaquine regimen. Capillary plasma concentration-time profiles were characterized using nonlinear mixed-effects modeling. The population pharmacokinetics of piperaquine were described accurately by a two-transit-compartment absorption model and a three-compartment distribution model. Body weight was a significant covariate affecting clearance and volume parameters. The individually predicted day 7 capillary plasma concentration of piperaquine was an important predictor (P < 0.0001) of recurrent malaria infection after treatment. Young children (2-5 years of age) received a significantly higher body weight-normalized dose than older children (P = 0.025) but had significantly lower day 7 piperaquine concentrations (P = 0.024) and total piperaquine exposures (P = 0.021), suggesting that an increased dose regimen for young children should be evaluated.

[Main characteristics of the street food sector in Bobo-Dioulasso, Burkina Faso]. / Caractéistiques de l'alimentation de rue dans la ville de Bobo-Dioulasso, Burkina Faso.

To investigate the sector of food sold in the streets of Bobo-Dioulasso and identify relevant information for action, a survey on knowledge and practices of street food vendors and consumers was conducted in June 2005. Data have been collected in 928 street food selling posts. Structured questionnaires were used to collect data from 874 street vendors and 2474 consumers. Street food sites are concentrated in places where administration and trade activities are usually running. The street food seller is a married and illiterate woman of 32 years old. Cereals (48.5%), meat (33.9%), milk (9.6%) and fruits (4.4%) are the basic consumables. The street food consumer is a non married man, 27 years old working in profit-making activity. Consumers use many criteria to choose the place to eat, at times or permanently. The street food sector represents a source of income and induces change in household eating habits. Street food in Bobo-Dioulasso needs to be better organised, by using an holistic approach that involves all the actors.

In vitro and in vivo antiplasmodial activity of 'saye', an herbal remedy used in Burkina Faso traditional medicine.

'Saye', a traditional medicine used in Burkina Faso, which consists of extracts of Cochlospermum planchonii (rhizome), Cassia alata (leaf) and Phyllanthus amarus (whole plant), showed a significant effect against Plasmodium falciparum and Plasmodium berghei parasites grown in vivo (IC(50) = 80.11 +/- 3.40 microg/mL; ED(50) = 112.78 +/- 32.32 mg/kg). In vitro the activity was lower.

[An action research network to improve the quality of tuberculosis care in West Africa]. / Améliorer la qualité des soins aux patients tuberculeux par la recherche-action en réseau. Une expérience en Afrique de l'Ouest.

BACKGROUND: Improvement in management systems for tuberculosis (TB) care is urgently needed in West Africa. In 2003, an experimental action research network began there, involving care providers, health system managers, and TB programme managers. Each project in all 6 countries used a "patient-centered" approach to improve tuberculosis case management. METHODS: The research teams included care providers, district medical officers, anthropologists and TB programme managers. Each research team conducted its project for a one-year period and then assessed its results. The specific problems identified were low TB detection rates (Burkina Faso, Côte d'Ivoire and Niger) and poor compliance among patients receiving treatment, including their ensuing loss to follow-up (Benin, Mali and Senegal). Investigators concluded that these weaknesses were due to the lack of access to care (geographical, financial and cultural), the complexity of the care system and the low quality of care. Solutions for all 6 countries aimed at improving access to high-quality care. RESULTS: One year after the experiment began, results varied from one country to another. In general, all participants understood the need to collaborate beyond national health systems because the problems from all 6 countries were quite similar. The research process led to better sharing of work between care providers and sometimes between care providers and TB patients. It provided participants with new concepts and a constant opportunity to implement them. These repeated meetings, however, keep care providers away from their offices. CONCLUSION: The research would have improved case management and care more effectively had the teams taken into account the psychological and sociological need of TB patients. A new regional dynamic has begun and must be pursued to help improve health care systems.

Molecular and antigenic evolution and geographical spread of H5N1 highly pathogenic avian influenza viruses in western Africa.

In Africa, highly pathogenic avian influenza H5N1 virus was first detected in northern Nigeria and later also in other regions of the country. Since then, seven other African countries have reported H5N1 infections. This study reports a comparison of full-length genomic sequences of H5N1 isolates from seven chicken farms in Nigeria and chicken and hooded vultures in Burkina Faso with earlier H5N1 outbreaks worldwide. In addition, the antigenicity of Nigerian H5N1 isolates was compared with earlier strains. All African strains clustered within three sublineages denominated A (south-west Nigeria, Niger), B (south-west Nigeria, Egypt, Djibouti) and C (northern Nigeria, Burkina Faso, Sudan, Côte d'Ivoire), with distinct nucleotide and amino acid signatures and distinct geographical distributions within Africa. Probable non-African ancestors within the west Asian/Russian/European lineage distinct from the south-east Asian lineages were identified for each sublineage. All reported human cases in Africa were caused by sublineage B. Substitution rates were calculated on the basis of sequences from 11 strains from a single farm in south-west Nigeria. As H5N1 emerged essentially at the same time in the north and south-west of Nigeria, the substitution rates confirmed that the virus probably did not spread from the north to the south, given the observed sequence diversity, but that it entered the country via three independent introductions. The strains from Burkina Faso seemed to originate from northern Nigeria. At least two of the sublineages also circulated in Europe in 2006 as seen in Germany, further suggesting that the sublineages had already emerged outside of Africa and seemed to have followed the east African/west Asian and Black Sea/Mediterranean flyways of migratory birds.

Amodiaquine metabolism is impaired by common polymorphisms in CYP2C8: implications for malaria treatment in Africa.

Metabolism of the antimalarial drug amodiaquine (AQ) into its primary metabolite, N-desethylamodiaquine, is mediated by CYP2C8. We studied the frequency of CYP2C8 variants in 275 malaria-infected patients in Burkina Faso, the metabolism of AQ by CYP2C8 variants, and the impact of other drugs on AQ metabolism. The allele frequencies of CYP2C8*2 and CYP2C8*3 were 0.155 and 0.003, respectively. No evidence was seen for influence of CYP2C8 genotype on AQ efficacy or toxicity, but sample size limited these assessments. The variant most common in Africans, CYP2C8(*)2, showed defective metabolism of AQ (threefold higher K(m) and sixfold lower intrinsic clearance), and CYP2C8(*)3 had markedly decreased activity. Considering drugs likely to be coadministered with AQ, the antiretroviral drugs efavirenz, saquinavir, lopinavir, and tipranavir were potent CYP2C8 inhibitors at clinically relevant concentrations. Variable CYP2C8 activity owing to genetic variation and drug interactions may have important clinical implications for the efficacy and toxicity of AQ.

[In vivo sensitivity of Plasmodium faciparum to chloroquine and sulfadoxine pyrimethamine in the Bobo Dioulasso region (1998-2001): risk factors associated with treatments failures to the two drugs]. / Sensibilité in vivo de Plasmodium falciparum à la chloroquine et à la sulfadoxine-pyrimethamine dans la région de Bobo Dioulasso (1998-2001): étude des facteurs de risque associés aux échecs thérapeutiques de ces deux médicaments.

The therapeutic efficacy of chloroquine (CQ) and sulfadoxine-pyrimethamine (SP) was determined over a 4 year period (1998-2001) in Bobo Dioulasso, Burkina Faso, with an analysis of the risk factors associated to treatment failures to the 2 drugs. In total, 2008 children (6 months-15 years old) attending in 4 health centres (1 urban and 3 rural) were included in the study. Children were alternatively allocated to either CQ or SP The WHO 14-days in vivo field test was carried out. PCV was measured at day 0 and 14. CQ treatment failure was 24.4% (229/940), most of them being late failures. Between 1998 and 2001 a significant increase in CQ treatment failure (p < 0.001) was observed. SP showed a good efficacy with a total treatment failure of 4.4% (33/749). However; a significant increase of resistance to this drug (p=0.001) was also observed between 1998 and 2001. Among children with anaemia at day 0.85% (23/27) were no more anaemic by day 14 in the SP group, while in the CQ group the proportion was lower; 69% (27/39). However the difference between the two drugs was not significant (p > 0.1). Univariate analysis showed that the site, the age of children, the time of recruitment and the parasitaemia were significantly associated with CQ treatment failure. In the multivariate analysis these 4 variables remain significantly and independently associated with the risk of CQ treatment failure. After adjusting for the effect of the 3 other factors, the risk of treatment failure was reduced by half in rural area compared to urban area as well as in children of 5-15 years of age compared to those under 5. The risk of treatment failure was significantly increased in 2000-2001 (OR = 1.66, p < 0.05) as compared to the 2 previous years (1998-1999). It was also twice higher in children with parasitaemia > or = 16,000/microl than in those having a lower parasitaemia. For SP we have not observed such connexions with the univariate and multivariate analysis.

Decentralising tuberculosis case management in two districts of Burkina Faso.

SETTING: In West Africa, national tuberculosis programmes (NTPs) face many problems due to the low performance of health care delivery systems and patients' social and cultural environment. OBJECTIVE: To improve the case management of TB in Burkina Faso. DESIGN: Using the operational research process as a tool, TB case management was decentralised from the district hospital to eight primary health care centres in 2003. RESULTS: Twelve months after decentralisation, the quality of case detection remained satisfactory. The delay between the identification of TB suspects with chronic cough and the confirmation of TB was reduced from 13 to 6 days. The detection rate of TB suspects during the study (30%) was twice as high as for 2001 and 2002 (15%). However, the detection rate for smear-positive TB cases decreased from 32.3% in 2001 and 2002 to 6.5% during the year of the study. CONCLUSION: Sufficient time and commitment are essential to obtain a case management system that is decentralised and effective. Efforts therefore need to continue to obtain more information and better results.

Enzyme-linked immunosorbent assay with worm vomit and cercarial secretions of Schistosoma mansoni to detect infections in an endemic focus of Burkina Faso.

Cercariae and adult Schistosoma mansoni were used to prepare, respectively, cercarial secretions (CS) and worm vomit (WoV). These were used as antigens in an enzyme-linked immunosorbent assay (ELISA) to test the IgG-reactivity of sera obtained in an S. mansoni-endemic area of Burkina Faso. Among the egg-excreting individuals (n = 240), 94.6% reacted positively with WoV, but only 62.9% with CS, thus suggesting a high diagnostic sensitivity of WoV, but not of CS. Among those individuals without detectable eggs in two Kato-Katz thick smears from different stool specimens (n = 215), the respective percentages of positive IgG reactivity were 78.1% and 63.3%. These positive reactions in the absence of detectable eggs are interpreted in terms of limited sensitivity of parasitological stool examinations. Optical density values in ELISA with CS, but not with WoV, correlated negatively with age, which may reflect decreasing exposure to cercariae in older individuals.

First report of a kdr mutation in Anopheles arabiensis from Burkina Faso, West Africa.

The leu-phe kdr mutation was detected in a specimen of Anopheles arabiensis during an extensive survey of pyrethroid resistance in An. gambiae s.l. in Burkina Faso. The detection of this mutation in An. arabiensis, which had so far been observed only in An. gambiae s.s., is important at both epidemiologic and fundamental levels. It can be useful to understand the history of this gene throughout the range of An. gambiae complex.

Natural swarming behaviour of the molecular M form of Anopheles gambiae.

In Anopheles gambiae, as in most species of mosquitoes, mating is initiated in flight. The males aggregate in aerial swarms and conspecific females individually fly to these swarms where they mate with males. In this study, we investigated the swarming behaviour of A. gambiae and conducted 2 surveys in the rice field area of the Vallée du Kou in Burkina Faso in 1999 and 2002. A high number of anopheline mosquitoes were observed in this area and both molecular M and S forms of A. gambiae were found in sympatry. Swarms formed a few minutes after sunset in different places and no obvious markers were associated with their occurrence. However, swarms occurred close to cow herds generally in open flat areas, 2-3 m above the ground. Overall, 2829 anopheline mosquitoes were collected from 21 swarms composed primarily of males. A few specimens of Culex quinquefasciatus were collected from 3 swarms. Although both molecular M and S forms were found in sympatry in the village, swarms were composed almost exclusively of the molecular M form. This suggests that there are alternative swarming habits for both molecular M and S forms of A. gambiae in nature.

Malaria: use of restriction endonuclease digestion and mutation-specific PCR for antifolate resistance isolate detection.

Antifolate resistance isolates of Plasmodium falciparum in the blood of 56 patients was investigated by using PCR technology. DNA was extracted with three different methods from parasite lysate by phenol-chloroform, or from whole blood and from blood collected onto dry filter paper, by chelex-100. The expected 727-bp PCR product was obtained in all samples extracted by chelex-100, while three samples prepared by phenol-chloroform failed to show any amplified product. The crucial point mutation within the dhfr gene leading to pyrimethamine and cycloguanil resistance is localised in an Alul recognition site. Thus, the 727-bp PCR product was submitted to endonuclease digestion. Fifty out of the 56 blood samples analysed yielded the two expected restriction fragments and an undigested 727-bp band. These 50 samples likely represent mixed infection as also confirmed the specific mutation PCR. The six undigested samples amplify a 339-bp fragment using a nested PCR-specific for pyrimethamine resistance mutation. Our results show that, the rapid DNA extraction from blood using chelex-100 and the PCR endonuclease assay can be efficiently used for accurate chemosensitivity analysis in the field.

Chloroquine and sulphadoxine-pyrimethamine efficacy for uncomplicated malaria treatment and haematological recovery in children in Bobo-Dioulasso, Burkina Faso during a 3-year period 1998-2000.

We determined the parasitological resistance and the clinical failure to chloroquine (CQ) and sulphadoxine-pyrimethamine (SP) by the WHO 14-day in vivo test over three consecutive years in 948 children aged 6-59 months with uncomplicated malaria attending four health centres in the province of Houet, Burkina Faso. Children were alternatively allocated to either CQ or SP. Packed cell volume (PCV) was measured at days 0 and 14. Parasitological resistance (RI, RII and RIII) to CQ was 18% (83 of 455) and to SP <1% (two of 308). Clinical failure with CQ was 12% (53 of 455) with no evidence of increase over time. Only one case of clinical failure was detected among the children treated with SP. The prevalence of anaemia (PCV <25%) was about 40% at day 0 and had decreased substantially by day 14 in both groups. However, in children treated with SP the prevalence of anaemia at day 14 was significantly lower than in those treated with CQ:RR = 3.15 (95% CI: 1.33-7.42, P = 0.008). CQ and SP are still efficacious for the treatment of uncomplicated malaria in children, at least in this area of Burkina Faso. However, the prevalences of CQ resistance reported from other areas of the country are worrying because of its potential spread. Regular surveillance of resistance to commonly used antimalarial drugs should continue.

[Preliminary study of cutaneous leishmaniasis in the town of Ouagadougou from 1996 to 1998]. / Etude préliminaire de la leishmaniose cutanée dans la ville de Ouagadougou de 1996 à 1998.

Since 1996, there have been reports of cases of cutaneous leishmaniasis in the town of Ouagadougou. The incidence has been on the rise but precise figures are not known. The object of the present study has been, first, to record cases of cutaneous leishmaniasis having occurred in private and public health centres in Ouagadougou from 1996 to 1998 and, second, to determine the progression of the disease in space and time. We wished also to confirm clinical cases in 1998 by parasitological examination, identify different clinical forms of the disease and map out cases in the town. We carried out a retrospective study from 1996 to 1998 and a prospective study in 1998. All cases recorded in this period in visited health centres were included. A total of 1845 cases of cutaneous leishmaniasis was identified, 50.3% of whom concerned women. The age of patients varied between 1 and 79 years for 356 patients, with a mean age of 26.7 years. Cases increased between 1996 and 1998 (1996 = 61 cases, 1997 = 552 cases, 1998 = 1218 cases). The months of highest incidence were August (13%), September (15%) and October (17%). Peripheral districts (28, 30, 29, 16, 15) in south-eastern areas of the town were the worst touched with 87% of cases. On average, patients seek care after 2 months of progression of the disease. The ulcero-crusted form (68.2%) was the most frequent clinical form observed for 327 patients, but almost half of the cases had more than one site of infection, (43.5%). Over half of the patients presented fewer than 10 lesions with an average of 6. The most common locations were on uncovered parts of the body, notably the superior (53%) and inferior limbs (49%). The parasite could be tested for by smear on 52 patients only in 1998 and 53.8% of cases tested were positive. Leishmania major, which is very prevalent in West Africa was identified in one patient. The vectors and main reservoirs of the parasite were not studied. Case management was generally incomplete; the most commonly prescribed drugs were antibiotics (70% of patients), but self-medication was frequent. Our recommendations after this preliminary study are: undertake multidisciplinary studies on cutaneous leishmaniasis in Ouagadougou in order to understand the local aetiology (vectors responsible for transmission, rodent and domestic animals involved in the epidemiological chain, parasite species); identify all other areas in the country where the disease is highly prevalent provide health care staff with a decisional algorithm and protocol therapy carry out and active control programme for cutaneous leishmaniasis in Burkina Faso.

[In vitro susceptibility of 232 isolates of Plasmodium falciparum to antimalarials in Burkina Faso (West Africa)]. / Etude de la sensibilité in vitro de 232 isolats de Plasmodium falciparum aux antipaludéens au Burkina Faso (Afrique de l'Ouest).

Plasmodium falciparum in vitro susceptibility to chloroquine, quinine, mefloquine and halofantrine was investigated in patients living in Bobo-Dioulasso (Burkina Faso, West Africa). Our study was carried out from July to November 1997 at the Malaria Chemoresistance Reference Centre, Centre MurazIOCCGE. Inclusion criteria were: presence of a single infection by R falciparum with a parasite count > or =4000 infected red cells/mm3. The susceptibility to drugs was measured after an incubation period of 48 hours at 37 degrees C, under 5% CO2. (3H) Hypoxanthine was added to the medium to monitor parasite growth. 134 isolates of P. falciparum were tested against chloroquine; 24.6% (33/134) were resistant. We have also documented 11.2% (15/133) of resistant isolates to halofantrine. All the tested isolates were susceptible to quinine (n=135) and mefloquine (n=136). A significant positive correlation was found between the following IC50 values: chloroquine-quinine, quinine-mefloquine and mefloquine-halofantrine. Our study shows no significant increase of the prevalence of chloroquine-resistant strains of P. falciparum in our study area; as well as the persistence of resistance to halofantrine with regard to previous publications in the subject.

[The simplified isotopic microtest: a method for studying the drug resistance in vitro of Plasmodium falciparum to antimalarial drugs]. / Le microtest isotopique simplifié: une méthode pour l'étude de la chimio-résistance in vitro de Plasmodium falciparum aux antipaludiques.

We evaluated the efficiency of the simplified version of the isotopic microtest (simplified test) and compared it to that of the complete version (standard test), for determining the susceptibility of local strains of Plasmodium falciparum to chloroquine, quinine, mefloquine and halofantrine. The study was carried out from July to November 1996, at the MURAZ Center, Bobo-Dioulasso, Burkina Faso. The inclusion criteria were: single infection with P. falciparum, with a parasite count of at least 4,000 infected red blood cells per mm3. Susceptibility to each drug was determined after incubation for 48 hours at 37

In vitro sensitivity of Plasmodium falciparum to halofantrine compared with chloroquine, quinine and mefloquine in the region of Bobo-Dioulasso, Burkina Faso (West Africa).

The in vitro sensitivity of P. falciparum drug-resistant isolates was evaluated in the region of Bobo-Dioulasso during the 1995 and 1996 rainy seasons. Two routinely used antimalarials (chloroquine and quinine) and two new antimalarials (mefloquine and halofantrine) were assessed using 24-hour in vitro cultures with tritiated hypoxanthine and a parasite density > or = 4,000/microl of blood. The proportion of chloroquine-resistant isolates was 20% in 1995 and 19% in 1996, whilst in 1996, the proportion of isolates resistant to halofantrine was greater than in 1995 (9.6% versus 1%). No significant differences were seen in the mean IC50 values in relation to the susceptibility of chloroquine-resistant or chloroquine-sensitive isolates to mefloquine and halofantrine. In the case of quinine, the mean IC50 values were significantly higher in chloroquine-resistant isolates than in chloroquine-sensitive ones. A significant positive correlation was found between the following IC50 values: chloroquine versus quinine, quinine versus mefloquine and mefloquine versus halofantrine.

[An outline of a method for estimating the calculated economic cost of malaria cases: its application to a rural area in Burkina Faso (Western Africa)]. / Esquisse d'une méthode d'estimation du coût économique chiffré des accès palustres: application à une zone rurale au Burkina Faso (Afrique de l'Ouest).

A precise method to estimate the cost of malaria in a rural area was developed and applied in 3 villages in Burkina Faso. The estimate takes into account direct costs such as consultation fees, microscopic examinations, medication and transport as well as indirect costs caused by lost work-days. The formula uses 6 variables: age of subject, degree of invalidity, duration of illness, profession, income and percentage of income lost. In the region of Bobo-Dioulasso, 3065 health centre clients were registered in the course of the study: 17% had been clinically diagnosed as having malaria but this was confirmed microscopically in only 11.6% of cases; 73.1% were children aged < 5 years, 13.9% children aged 6-15 years, 12.2% adults aged 16-50 years and 0.8% adults aged > 50 years. Most patients worked in agriculture and trade. The average duration of illness was 4 days, with each case incurring a cost of $11.7 comprised of $8 direct costs and $3.7 indirect costs.