First Report of Fludioxonil Resistance in Botrytis cinerea from a Blackberry Field in Georgia.

Botrytis cinerea Pers. is an important plant-pathogenic fungi responsible for gray mold on more than 230 plant species worldwide, including blackberry (Rubus). One of the main strategies to control the disease involves the application of different classes of fungicides. The phenylpyrrole fludioxonil is currently marketed in combination with the anilinopyrimidine cyprodinil as Switch 62.5WG (Syngenta Crop Protection Inc., Greensboro, NC) for gray mold control. In August 2013, blackberries affected with symptoms resembling gray mold were collected from a field located in Berrien County (Georgia), where Switch 62.5WG had been used extensively over the last 5 years. Three single-spore isolates, each from a different fruit, were obtained and identified as B. cinerea on the basis of morphology and confirmed by a 238-bp PCR amplification product obtained with primer set G3PDH-F1 (5'-GGACCCGAGCTAATTTATGTCACGT-3'), G3PDH-F2 (5'-GGGTGTCAACAACGAGACCTACACT-3'), and G3PDH-R (5'-ACCGGTGCTCGATGGGATGAT-3'). In vitro sensitivity to fludioxonil (Scholar SC, Syngenta) was determined on 1% malt extract agar (MEA) using a conidial germination assay as previously described (4). One isolate was moderately resistant due to growth on medium amended with the discriminatory dose of 0.1 µg/ml fludioxonil and residual growth at 10 µg/ml (4). To assess performance of fludioxonil in detached fruit assays, commercially grown strawberries (24 in total for each isolate and treatment) were rinsed with water, dried, and sprayed 4 h prior to inoculation with either water (control fruit) or 2.5 ml/liter of Scholar SC to runoff using a hand mister. Scholar SC was used because fludioxonil was the sole active ingredient in this product and strawberries were used because latent infections in fresh blackberry fruit interfered with inoculation experiments. This dose reflects the rate recommended for postharvest gray mold control according to the Scholar label. Fruit was stab-wounded with a sterile syringe and inoculated with a 30-µl droplet of conidia suspension (106 spores/ml) of the two sensitive or the resistant isolate. After inoculation, the fruit were kept at 22°C for 4 days. The sensitive isolates developed gray mold on non-treated (2.7 cm lesion diameter) but not on Scholar SC-treated fruit (0.0 cm lesion diameter). The resistant isolate developed gray mold disease on the water-treated control fruit (2.5 cm lesion diameter) and the fungicide-treated fruit (1.8 cm lesion diameter). EC50 values were determined in microtiter assays as described previously (3) using the concentrations of 0.01, 0.04, 0.12, 0.37, 1.1, 3.3, and 10 µg/ml fludioxonil. Values were 0.02 and 0.05 µg/ml for the two sensitive isolates and 3.15 µg/ml for the resistant isolate. All experiments were performed twice. This is the first report of fludioxonil resistance in B. cinerea from blackberry in Georgia. Prior to this study, resistance to fludioxonil in B. cinerea was reported in France, Germany, and only a few states in the United States including Maryland, South Carolina, Virginia, and Washington (1,2). The emergence of resistance to fludioxonil emphasizes the importance of resistance management strategies. References: (1) D. Fernández-Ortuño et al. Plant Dis. 97:848, 2013. (2) D. Fernández-Ortuño et al. Plant Dis. 98:692, 2013. (3) M. Kretschmer et al. PLOS Pathog. 5:e1000696, 2009. (4) R. W. S. Weber and M. Hahn. J. Plant Dis. Prot. 118:17, 2011.

Altered free radical metabolism in acute mountain sickness: implications for dynamic cerebral autoregulation and blood-brain barrier function.

We tested the hypothesis that dynamic cerebral autoregulation (CA) and blood-brain barrier (BBB) function would be compromised in acute mountain sickness (AMS) subsequent to a hypoxia-mediated alteration in systemic free radical metabolism. Eighteen male lowlanders were examined in normoxia (21% O(2)) and following 6 h passive exposure to hypoxia (12% O(2)). Blood flow velocity in the middle cerebral artery (MCAv) and mean arterial blood pressure (MAP) were measured for determination of CA following calculation of transfer function analysis and rate of regulation (RoR). Nine subjects developed clinical AMS (AMS+) and were more hypoxaemic relative to subjects without AMS (AMS-). A more marked increase in the venous concentration of the ascorbate radical (A(*-)), lipid hydroperoxides (LOOH) and increased susceptibility of low-density lipoprotein (LDL) to oxidation was observed during hypoxia in AMS+ (P < 0.05 versus AMS-). Despite a general decline in total nitric oxide (NO) in hypoxia (P < 0.05 versus normoxia), the normoxic baseline plasma and red blood cell (RBC) NO metabolite pool was lower in AMS+ with normalization observed during hypoxia (P < 0.05 versus AMS-). CA was selectively impaired in AMS+ as indicated both by an increase in the low-frequency (0.07-0.20 Hz) transfer function gain and decrease in RoR (P < 0.05 versus AMS-). However, there was no evidence for cerebral hyper-perfusion, BBB disruption or neuronal-parenchymal damage as indicated by a lack of change in MCAv, S100beta and neuron-specific enolase. In conclusion, these findings suggest that AMS is associated with altered redox homeostasis and disordered CA independent of barrier disruption.

Aggregation effects on oxygen binding of sickle cell hemoglobin.

Deoxygenation of concentrated solutions (0.33 gram per milliliter) of sickle cell hemoglobin show (i) a "crisis point" where the oxygen binding curve is unusually steep (Hill coefficient of 5 to 6), and (ii) a simultaneous increase in light scattering. Nearly identical oxygen binding curves are obtained upon oxygenation and deoxygenation of these solutions. The influence of aggregation is to shift the curve toward higher pressures.

[DRESS syndrome secondary to pyrazinamide: An uncommon complication of tuberculosis treatment]. / DRESS syndrome au pyrazinamide : complication rare et grave du traitement antituberculeux.

INTRODUCTION: The Drug Reaction with Eosinophilia and Systemic Symptom (DRESS) is a severe drug-induced reaction. CASE REPORT: We report the case of a 35-year-old man treated by RHEZ for a first episode of a smear positive pulmonary tuberculosis and who developed a DRESS syndrome due to pyrazinamide after twenty days of treatment, associated with a viral reactivation to Human Herpes Virus 6 (HHV6). He had a skin eruption, liver involvement and hypereosinophilia. He fully recovered after drug withdrawal, associated with local and general corticosteroids. He died two weeks after discharge. CONCLUSIONS: Discovery of DRESS syndrome during tuberculosis treatment is an uncommon complication and requires a searching for the responsible drug. That should be difficult because tuberculosis drugs are often given as fixed-dose combination. Physicians have to bear in mind the potential role of pyrazinamide.

[Low level educated community health workers training: a strategy to improve children access to acute respiratory treatment in Senegal]. / La formation des agents de santé communautaire instruits: une stratégie pour améliorer l'accès des enfants au traitement des infections respiratoires aiguës au Sénégal.

BACKGROUND: Acute respiratory infections (ARI) are the leading cause of death in childhood. Most of children's deaths happen at home without health level first cares. However in Senegal only health workers are allowed to prescribe antibiotics. A competency-based training was developed to improve and assess the management of acute respiratory infections in young children aged 2 to 59 months by low level educated community health workers (CHWs) in 4 districts of Senegal. The first findings showed the CHWs capable of acquiring the skills needed to effectively manage ARI cases in accordance with the World Health Organization's ARI case management strategy. The aim of this study was to assess the quality of their management after a 1 year follow-up. METHODS: We provided to CHWs 3 days course in ARI management. After the 3-day course, a 4-month follow-up was performed. We organized a 1-day refresher course every month and in every district. In order to assess the quality of management of CHWs we analyzed the management process and compared the CHWs classification to the classification of the first level health facilities. RESULTS: Twenty-three percent (3727/15,965) of IRA cases of district were managed by CHWs. Ninety percent (2738/3042) among them were well classified, well managed and well followed-up. But 28% of severe pneumonia cases were misclassified as pneumonia. About treatment, 22.5% of 'cough or cold' got wrong treatment with cotrimoxazole and 10.3% of severe pneumonia got cotrimoxazole without referral. Less than half of severe pneumonia benefited of the first follow-up and only 18% of the second. CONCLUSIONS: The results highlight that a CHWs low-level educated in French, trained and followed-up could apply the WHO algorithm of IRA management. They could help to give proximal care related to children ARI, to the community. But it seems useful of emphasizing the recognition of danger signs and the follow-up of severe cases.

[Diabetic ketoacidosis of the child revealed by a pseudo-surgical acute abdomen. A case report]. / Acidocétose diabétique de l'enfant révélée par un tableau d'abdomen aigu pseudo-chirurgical: a propos d'un cas.

Ketoacidosis is a frequent mode of revelation of diabetes of the child. Signs of diabetes are generally unperceived or are badly interpreted by the family entourage. We report a case of diabetic ketoacidosis in an eight-year-old child without particular medical history. The context of discovery was an acute abdomen by gastric dilatation in a severe context of dehydration. Evolution was quickly favourable after medical treatment with disappearance of acute abdominal signs (clinical and radiological). This observation enables us to recall, by the light of a review of the literature, that diabetic ketoacidosis can be revealed by pseudo-surgical acute abdomen and treatment is purely medical.

Severe congestive heart failure and cardiomyopathy as a complication of myotonic dystrophy in pregnancy.

The pregnancy of a patient with myotonic dystrophy and heart failure due to cardiac involvement is described. Endomyocardial biopsy was performed at 32 weeks' gestation with echocardiographic guidance to establish the diagnosis. Severe congestive heart failure, refractory to conventional therapy, was encountered. Continuous arteriovenous hemofiltration was used to relieve pulmonary edema before cesarean delivery.

The effect of non-binding molecules on the gelation of HbS.

The influence of an inert globular macromolecule upon the solubility of sickle cell hemoglobin has been determined as a function of the degree of oxygenation. The thermodynamic theory required to treat this and related problems is derived starting with the Gibbs-Duhem equation and introducing the effect of specific binding (oxygen) by use of the binding partition function. The treatment includes non-ideal solution behaviour as measured by osmotic pressure of highly concentrated macromolecular solutions. Application of the theoretical equation demonstrates how the solubility of hemoglobin is influenced by the presence of the binding ligand (oxygen) and the inert macromolecule, bovine serum albumin (BSA). Good agreement is obtained between experimentally determined and theoretically calculated solubilities using 1) oxygen binding curves to solution and gel phases, 2) activity coefficients from osmotic pressure data, 3) one solubility under the condition where oxygen and BSA are absent, and 4) the value of the water content of the gel phase. Examination of theoretical equations suggests that inert molecules of intermediate size, that are partially excluded from crystalline or gel phases, have the potential of generally increasing the solubility when non-ideal solution effects are small.

Striking similarities between HIV-1 Env protein and the apoptosis mediating cell surface antigen Fas. Role in the pathogenesis of AIDS.

We have designed a computer strategy in order to detect systematically peptidic sites with the potential of interfering with the immune regulatory processes. Applying this software to HIV-1 proteins has led us to unravel a few peptidic sites which could either act directly or be the targets of an auto-immune reaction during HIV-1 infection. We previously reported that the SLWDQ pentapeptide identity with a critical site of CD4 could trigger in HIV-1 infected individuals both an humoral and a cellular autoimmune reaction. In this study, we focused on surprising similitudes unravelled by our software Automat, between HIV-1/2 and another immunoregulatory molecule, the Fas protein which is also called the apoptosis-mediating cell-surface antigen.

HIV-1-induced immune suppression may result from autoimmune disorders including anti-SLWDQ autoantibodies.

We have previously unravelled the striking SLWDQ pentapeptide identity between HIV-1 env gp120 and the CD4 molecule. We show here that this pentapeptide is required for the functioning of the co-stimulatory MHC-CD4 signal in T4-cell activation since it suppresses antigen-induced T-cell proliferation. Moreover, concerning the MHC class II counterpart, the LNGQEETGVVSTN sequence which strongly inhibits T-cell immune activation is likely to be part of the functional site of the molecule. Interestingly the MHC/gp120 homology described by Young overlaps this MHC region. We further report that the gp120 SLWDQ peptide triggers an immune reaction which is both humoral (anti-SLWDQ antibodies) and cellular (CTLs against autologous targets carrying the pentapeptide) in HIV-1 infected individuals. Finally, anti-SLWDQ antibodies from patients sera purified by column chromatography strongly inhibit antigen-induced immune T-cell activation. This result led us to postulate that these antibodies found in high titers in HIV-1 infected individuals could contribute to set up the progressive systemic immune T-cell suppression characterizing AIDS.

Biological effect of active anti-IFN alpha immunization in HIV-infected patients.

Circulating interferon (IFN) was investigated in HIV-1 seropositive patients by measuring the IFN alpha antiviral effect in the serum. While serum of healthy seronegative individuals exhibits an antiviral effect, not due to IFNs, considered as background, serum of seropositive patients showed an additional antiviral effect due to the abnormal presence of IFN alpha. Increased titers of IFN alpha were found in the course of the HIV infection and seemed to correlate with the evolution of AIDS disease. Furthermore, patients immunized against IFN alpha had both stabilized CD4 cell count and decreased IFN alpha in their serum. HIV-1-infected patients also exhibited higher titers of natural anti-IFN antibodies than seronegative controls and the level of specific antibodies (Abs) markedly increased in immunized patients. Finally, serum from immunized patients, when compared to seronegative controls, exhibits an interferon neutralizing capacity.

Critical sites: a semantic approach to protein sequences. Application to the HIV-1 envelope molecule.

We have designed two software systems allowing the study of proteins through a comparison to those stored in data banks. The first one, "Automat", locates in a systematic manner all identities shared by a given protein and the proteins in a data bank. The second, "Critic" enables the selection of specific segments in a given molecule by comparing them with those gathered in a data bank. These sites were termed "critical" since they mostly correspond to functional sites (active sites) of the well-known proteins which were studied with the aid of this program (somatostatin, insulin, IL2, etc). Automat allowed us to reveal homologies between HIV-1 and the CD4, which have remained unsolved until now. These similitudes proved to be critical sites (according to Critic). The putative involvement of these sites in the physiopathological processes as induced by HIV-1 are worth considering since the results of our experiments are consistent with this assumption.

Removal of gp160 induced bio-hazards for a safe AIDS vaccine candidate.

In the first AIDS vaccine trial, immunizing preparations were based on HIV-1 Env protein (gp160). Immunogenic properties of gp160 which trigger both a humoral and cellular immune response have since justified its use in various vaccine programs, both past and present. Many reports however have underlined deleterious effects on the immune system--anti-HIV-1 enhanced antibodies, anti-CD4 autoantibodies, and inhibition of T cell activation by HIV-1--particularly associated with the Env protein. The present study shows that gp160 presented in a biologically inactivated but immunogenic form, as used in our trial, could avoid these complications. Bio-hazards associated with gp160 which indeed could be removed by appropriate treatment of the native protein, should be taken into consideration in AIDS vaccine programs.

Lovastatin inhibits HIV-1 expression in H9 human T lymphocytes cultured in cholesterol-poor medium.

The effects of the HMG-Coenzyme A reductase inhibitor lovastatin on HIV-1 expression and sterol synthesis have been investigated in the human H9 lymphocytic cell line. To this purpose, sterol synthesis from 14C-acetate, cell multiplication and reverse transcriptase activity have been measured in parallel at various times after cell infection by HIV-1. It was found that nine days after viral loading, lovastatin inhibited both sterol synthesis and viral multiplication as assessed by the reverse transcriptase activity. Since HIV infection has been shown to induce alterations in membrane cholesterol content, suggesting that the virus cycle may be partially dependent upon cellular cholesterol, inhibitors of cholesterol synthesis could be an interesting way of research in order to slower HIV propagation.

[Hyperthyroidism in people over 50 years of age in Senegal. Study of 31 cases observed over a 14-year period]. / Hyperthyroïdie après 50 ans en milieu sénégalais. Etude de 31 cas colligés en 14 ans.

PURPOSE: Though common in elderly, with poor and masked symptomatology, hyperthyroidism has never been studied in people older than 50 years of age in African countries. METHODS: Of the 300 cases of hyperthyroidism that were observed during this study, 31 belonged to this category. We analyzed the frequency of various parameters, demographic characteristics, motivation for consulting, thyrotoxic syndrome, non thyrotoxic signs, cardiac manifestations, hormonal and scintigraphic characteristics, etiology and etiologic factors, and treatment. RESULTS: The frequency of hyperthyroidism in this population was 10%. Housewives were more numerous, with 20 cases observed in the 31 patients. The main group (13/31) was of rural origin. Signs that predominantly led to consultation were weight loss (23/31), cervical tumor (17/31), and palpitations (12/31). Three major signs were associated with the thyrotoxic syndrome: weight loss (29 cases), tachycardia (27/31), and the existence of tremors in the extremities (22/31). Hormone assays showed that thyroxine (T4) was about 265 +/- 74 nmol/L and triiodothyronine (T3) about 6 +/- 2 nmol/L, at immunoradiometric assay; thyroid-stimulating hormone (TSH) was about 0.17 +/- 0.23 muIU/mL. Eye protrusion predominated in the nonthyrotoxic syndrome, with 25 cases in the 31 patients. Etiologic forms of the disease were composed of 25 Grave's disease, with 22 typical cases. Etiologic factors were various, however without any case of neoplasia. Cardiac complications included two cases of atrial fibrillation. No iatrogenic form of the disease was observed. Mean initial carbimazole dosage was about 34 +/- 8 mg/d. Of the 23 patients, 15 had a favourable outcome. CONCLUSIONS: In this series, the high number of housewives and the patients' rural origin were less characteristic of hyperthyroidism than the Grave's disease etiology. This might be due to the young age of this African population.

[Development of African children: influence of the home environment]. / Développement de l'enfant Africain: influence du milieu de vie.

The purpose of this report was to show how special features of the African home environment affect child development. Special emphasis was placed on the traditional social structure and the roles of various players. The different stages of development are described in relation to the major social events in African families and communities. Psychomotor development in African children presents several particularities in relation to children in the industrialized world but differences tend to decrease as a result of ongoing social upheaval. Modernization or globalization has had a profound effect on family organization and child care. These changes must be taken into account by officials in charge of child-psychiatry services. Preventive initiatives should taken into account and, insofar as possible, preserve traditional familial resources.

[Clinical characteristics of primary hypothyroidism in Dakar. Apropos of 37 cases]. / Caractéristiques cliniques de l'hypothyroïdie primaire à Dakar. A propos de 37 observations.

Primary hypothyroidism, other than cases of endemic goiter, has rarely been described in Africa. We conducted a retrospective study of the patients admitted to our hospital unit between 1985 and 1996. The inclusion criteria were clinical signs of hypothyroidism and low levels of thyroid-stimulating hormone. We investigated socio-demographic, clinical (hypometabolic syndrome, cutaneomucal syndrome, muscular syndrome) and etiological (spontaneous thyroid atrophy, thyroidectomy, multinodular goiter) factors. Overall, our study population contained 37 cases, 8 men and 29 women. The mean age of the men was 40.8 +/- 19.2 years and that of the women was 41.5 +/- 14.5 years. Eighteen patients (about 50%) lived in the suburbs, 25% of patients were from urban areas and 25% from rural areas. The associated clinical signs were: 1) hypometabolism: constipation (51% of cases), bradycardia (45%), physical asthenia (40%), sleeping during the day (32%), frilosity (35%); 2) cutaneomucal syndrome: hoarseness (48%), alopecia (32%), facial puffiness (27%), macroglossia (24%), hypoacousia (21%), weight gain (18%), dry skin (16%), pallor (2%); 3) muscular syndrome was rare: myalgia (4 cases), muscle weakness (2 cases). Mean total cholesterol concentration was 2.54 +/- 0.75 g/l; mean total T3 was 1.027 +/- 0.84 nmol/l; mean total T4 was 16.70 +/- 16.89 nmol/l; mean TSH concentration, measured by radiometry, was 63.74 +/- 51.01 mIU/l. The etiology was goiter in 13 cases, thyroidectomy (11 cases) and spontaneous thyroid atrophy (13 cases). Thus, primary hypothyroidism does occur in African hospitals, particularly in Senegal. This disease, which has traditionally been reported in public health studies of endemic goiter, also occurs in cosmopolitan African environments.

[Infections in Senegalese children and adolescents with sickle cell anemia: epidemiological aspects]. / Infections chez l'enfant et l'adolescent drépanocytaires sénégalais: aspects épidémiologiques.

Infection is the main factor of morbidity and mortality in children with sickle cell disease (SCD). The objective of this study is to determine it's epidemiologic outline in senegalese children and adolescents with SCD. We retrospectively studied infection data in all the charts of a cohort of 323 patients with SCD (307 SS, 13 SC and 3 s beta + thalassemia) followed at Albert Royer children hospital from january 1991 to december 1997. Serum sampling was systematically made for HIV and antigen HBs serology in all patients we received in the last 3 months (october to december 1997). Patients were aged from 5 months to 22 years (medium age = 8 years). 813 infection episodes were diagnosed, concerning 184 patients (56 per cent). SS patients were more affected (59 per cent) than the others (23 per cent, p = 0.04). ENT and broncho-pulmonary onsets were more frequent but had a generally benign course. Menigitidis, septicemia and osteomyelitis were exclusively diagnosed in SS patients. Their prevalences in this group were respectively: 1.0 per cent, 4.9 per cent and 9.8 per cent. HIV serology was determined in 155 patients, including 41 per cent with blood transfusion antecedents. All tests were negative. HBs antigen was determined in 104 patients and seroprevalence was 7.7 per cent in the whole group and 6.0 per cent in patients with transfusion antecedents and 7.7 per cent for the others. Plasmodium falciparum malaria onset was observed in 9.6 per cent of our patients and there was no case of cerebral malaria. Infection was involved in 9 of the 11 cases of death. Then infection constitute the major problem in children and adolescents with SCD in Dakar. However prevalences of severe onsets are comparable to data in Europe despite our poor follow up conditions. Senegal haplotype may lead to a good tolerance of SCD. Negative HIV serology and low HBs antigen seroprevalence in transfused patients are attributed to a relatively low level of HIV prevalence in the general population and a good transfusion security in Senegal.

[Acute encephalic manifestations in Senegalese children with sickle cell disease]. / Manifestations encéphaliques aiguës chez l'enfant drépanocytaire sénégalais.

The course of sickle cell disease (SCD) may be complicated by neurologic events, mainly bactérial meningitidis and stroke. We retrospectively studied all cases with acute encephalic manifestations (AEM) in a cohort of 461 children and adolescents with SCD followed at Albert Royer Children Hospital of Dakar (Senegal) from january 1991 to december 2000 (ten years). Among them 438 had sickle cell anemia (SCA), 19 SC disease and 4 S-beta thalassemia (3 S-beta+, 1 S-beta0). Seven patients, all with SCA, presented antecedents of AEM revealed by flacid and proportionnal hemiplegia evoking stroke. Prevalence of these AEM was 1.5 per cent among patients with SCD and 1.6 per cent among those with SCA. They were 4 girls and 3 boys (sex ratio = 0.75) aged 4 to 8.5 years when occurred the first accident. We observed no clinical or biological distinctive characteristic of SCA in these patients compared to those without crebrovascular accident. Recurrence was observed once in a boy after a 12 months interval and twice in a girl after 20 and 60 months intervals successively. No transfusionnal program was applied to prevent recurrent stroke because of insufficient conditions for long-term transfusion. Stroke appears to be rare in senegalese children with SCD. However it poses in our context the major problem of applicability of transfusionnal program which constitute the only therapy universally recognised to be effective to prevent recurrence. Nevertheless hydroxyurea could be a satisfactory alternative.