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The different aspects of treatment of periodontal disease and mucogingival defects all require an accurate diagnosis in addition to good control and precision during therapeutic procedures. Magnification aids and microsurgery, combined with minimally invasive techniques, can best meet these requirements. The suitability of treatment, the healing time, pain levels and postoperative scarring are all improved and the patient benefits.

With the increasingly commercial emphasis on dental esthetics, patients have become more interested in improving the appearance of their smile. For many, whiter teeth is their chief goal. However, when considering dental bleaching, practitioners need to take certain precautions. Over the past decade, multiple studies have evaluated the effects of bleaching agents on restorative materials. This article reviews their conclusions, focusing mainly on the clinical impact these agents can have on amalgam, porcelain, ormocer, glass ionomer, compomer and composite resin restorations.

Diagnostic systems for Lassa fever (LF), a viral hemorrhagic fever caused by Lassa virus (LASV), such as enzyme immunoassays for the detection of LASV antibodies and LASV antigens, were developed using the recombinant nucleoprotein (rNP) of LASV (LASV-rNP). The LASV-rNP was expressed in a recombinant baculovirus system. LASV-rNP was used as an antigen in the detection of LASV-antibodies and as an immunogen for the production of monoclonal antibodies. The LASV-rNP was also expressed in HeLa cells by transfection with the expression vector encoding cDNA of the LASV-NP gene. An immunoglobulin G enzyme-linked immunosorbent assay (ELISA) using LASV-rNP and an indirect immunofluorescence assay using LASV-rNP-expressing HeLa cells were confirmed to have high sensitivity and specificity in the detection of LASV-antibodies. A novel monoclonal antibody to LASV-rNP, monoclonal antibody 4A5, was established. A sandwich antigen capture (Ag-capture) ELISA using the monoclonal antibody and an anti-LASV-rNP rabbit serum as capture and detection antibodies, respectively, was then developed. Authentic LASV nucleoprotein in serum samples collected from hamsters experimentally infected with LASV was detected by the Ag-capture ELISA. The Ag-capture ELISA specifically detected LASV-rNP but not the rNPs of lymphocytic choriomeningitis virus or Junin virus. The sensitivity of the Ag-capture ELISA in detecting LASV antigens was comparable to that of reverse transcription-PCR in detecting LASV RNA. These LASV rNP-based diagnostics were confirmed to be useful in the diagnosis of LF even in institutes without a high containment laboratory, since the antigens can be prepared without manipulation of the infectious viruses.

Since forty years Marburg and Ebola viruses emerge frequently in Africa and are responsible of viral hemorragic fever outbreaks with high mortality rate. Despite intensive research programs, these viruses remain mysterious: the reservoir is not clearly defined, and the mechanisms leading to their high pathogenicity are poorly understood; a defective or inadapted immune response seems to be the main factor. No specific treatment nor vaccine are available for humans. But encouraging results have been obtained in the treatment of filovirus infections in non human primate model with different products, as recombinant nematode anticoagulant protein, anti sens phosphorodiamidate morpholino oligomers or small interfering RNA.As vaccines, recombinantVSV expressing the GP of filovirus or adenovirus expressing the GP and NP of filovirus are very promising in macaque models.

There have recently been large outbreaks of Marburg hemorrhagic fever (MHF) caused by Marburgvirus (MARV) in the Democratic Republic of Congo and Angola. The development of reliable diagnostic systems for MHF is urgently needed. An antigen-capture enzyme-linked immunosorbent assay (Ag-capture ELISA) using either of the two monoclonal antibodies (2A7 and 2H6) produced by immunizing mice with recombinant nucleoprotein of MARV was described (Journal of Medical Virology, 76, 111-118, 2005). In the present study, it was revealed that the Ag-capture ELISA specifically detected authentic MARV antigen and that the sensitivity of the Ag-capture ELISA was at a level similar to that of reverse-transcription polymerase chain reaction. These results suggest that the Ag-capture ELISA using the monoclonal antibodies, 2A7 and 2H6, is applicable to the diagnosis of MHF.

Background: Limitation of resident work hours has created the need to explore alternatives to surgeon presence during initial assessment and resuscitation for selected life-threatening injuries in children. We recently eliminated the requirement for surgeon presence during Level II alerts. The purpose of this study was to evaluate the impact of this change on patient care., Methods: A retrospective analysis of trauma alert activity was performed using data from our trauma registry. In March 2003, responsibility for level II alerts was transferred from the pediatric surgeons (PSs) to the Emergency Department (ED) physicians. We compared the activity in the 18-month period before this change (period 1; n = 627) to that afterward (period 2; n = 587). Outcome measures included injury severity score, emergency department length of stay, missed injuries, abdominal computed tomography use, and mortality. Data were analyzed using log-rank statistic, chi2, or t test, where appropriate, with significance level at P < .05., Results: During the entire study period, 1499 patients met the trauma alert activation criteria of which 1214 (81%) were level II alerts. The mean injury severity score for period 1 (8.5 +/- 7.3 SD) was similar to period 2 (9.0 +/- 7.1 SD). When ED physicians replaced PS for Level II alerts, ED length of stay increased from 135 minutes to 165 minutes (P < .001). In addition, the use of abdominal computed tomography was significantly decreased (53.6% vs 42.6%; P < .001). However, there were no missed injuries and no significant differences in the rate of mortality., Conclusions: When ED physicians replaced PS for Level II alerts, trauma room length of stay was increased, but use of abdominal imaging was decreased with no differences in rate of missed injury or mortality. Emergency Department physicians can safely replace PS during Level II alerts. These findings may be useful to institutions experiencing surgical workforce limitations for trauma alerts.

This study evaluated the surface finish of three direct resin composites polished with three different systems. Disk-shaped specimens (n=16 per material; phi=8.0 mm x h=2.0 mm) were formed in a stainless steel mold by packing uncured material, either a hybrid composite (Z250, 3M ESPE) or two micro-hybrid composites (Point 4, Kerr; Esthet-X, Dentsply), and light-cured from the top and the bottom surfaces with a light-emitting diode (LED) curing unit (NRG, Dentsply). After storing the specimens in deionized water at 37 degrees C for seven days, one side of each specimen was finished through 1200-grit SiC abrasive (Buehler). Five specimens of each resin composite were randomly assigned to one of the three polishing systems (Identoflex, Kerr; Pogo, Dentsply; Sof-Lex, 3M ESPE). Manufacturers' instructions were followed during the polishing procedures. The average surface roughness (Ra) was determined by generating tracings across the polished surface of each disk using a scanning profilometer (Surfanalyzer System 5000, Federal Products Co). The results were analyzed by Kruskal-Wallis and Mann and Whitney tests (p < or = 0.05). The smoothest surfaces were produced with the celluloid strip (control group) on all the resin composites tested. The aluminum oxide disks (Sof-Lex) produced a statistically equivalent surface finish (Ra) on the three resin composites. The lowest mean roughness values were recorded with diamond micropolisher disks (PoGo) on the hybrid composite (Z250). Overall, the two new polishing systems, Identoflex and PoGo, created a comparable surface finish to that produced by the Sof-Lex system on all three resin composites.

The recent emergence of monkey pox in the United States of America highlights the problem (known for other infectious agents) of dissemination of pathogens outside their endemic area, and of subsequent global threats of variable gravity according to agents. It is a real emergency since monkey pox had been confined to Africa for several decades, where small epidemics occurred from time to time, monkey pox is a "miniature smallpox" which, in Africa, evolves on an endemic (zoonotic) mode with, as reservoirs, several species of wild rodents (mainly squirrels) and some monkey species. It can be accidentally transmitted to man then develops as epidemics, sometimes leading to death. The virus was imported in 2003 in the United States of America, via Gambia rats and wild squirrels (all African species), and infected prairie dogs (which are now in fashion as pets), then crossed the species barrier to man. In the United States of America, screening campaigns, epidemiological investigations, and subsequent treatments led to a rapid control of the epidemic, which is a model of emergent disease for this country. Therapeutic and preventive measures directly applicable to monkey pox are discussed. They can also be applied against other pox virus infections (including smallpox). The risk of criminal introduction of pox viruses is discussed since it is, more than ever, a real worldwide threat.

Objectives: A potential problem with high-intensity lights might be failure of polymer chains to grow and cross-link in a desired fashion, thereby affecting the structure and properties of the polymers formed. The purpose of this study was to evaluate mechanical properties of resin composites polymerized using four different light-curing units., Methods: A conventional quartz-tungsten-halogen (QTH) light, a soft-start light, an argon-ion laser, and a plasma-arc curing light were used to polymerize disk-shaped (9.0mm diameter x 1.0 mm high) and cylinder-shaped (4mm diameter x 8 mm high) specimens of a universal hybrid and a flowable hybrid composite. Biaxial flexure strength, fracture toughness, hardness, compressive strength, and diametral tensile strength were determined for each composite., Results: The use of the plasma-arc curing light, a high-intensity light, resulted in significantly lower hardness for the universal hybrid composite compared with the hardness obtained using the conventional QTH and the soft-start units. Hardness was the only mechanical property that was adversely affected by the use of a high-intensity light., Significance: High-intensity lights might affect some resin composite mechanical properties, but this effect cannot be generalized to all resin composites and all properties.

Statement of Problem: intact, prepared, and restored human maxillary premolars., Material and Methods: Fifty intact, noncarious human maxillary premolars were divided into 5 groups of 10 and were mounted with their roots imbedded in autopolymerized acrylic. In the first group, the teeth were intact with no preparation. In the other 4 groups, Class II MOD preparations were made with a water-cooled high-speed hand piece. In 1 group, the cavity preparations were restored with bonded CAD/CAM ceramic inlays. In 2 groups, the preparations were restored with bonded CAD/CAM composite inlays (acid etched or air particle abraded). In the final group, the teeth were prepared but unrestored. Specimens were tested individually in a universal testing machine, in which a 4.82-mm-diameter steel sphere plunger was mounted in the crosshead moving at 0.5 mm/min. The plunger contacted the facial and lingual triangular ridges beyond the margins of the restorations. Peak load to fracture (N) was measured for each specimen. Means were calculated and analyzed with analysis of variance (P

Purpose: To evaluate microleakage at enamel and dentin margins of two composite resins, using bulk and incremental placement techniques, "rebonding", and facial and lingual curing methods., Materials and Methods: One hundred standardized Class V cavity preparations were made on the facial surface of extracted human premolars and were randomly assigned to 10 groups. Single Bond was used as the dentin/enamel adhesive. A heavily filled composite resin, Z250, and a microfill, Silux Plus, were inserted and polymerized using five different techniques: (1) incremental placement and facial curing; (2) incremental placement, facial curing and rebonding; (3) bulk placement and facial curing; (4) bulk placement, facial curing and rebonding; (5) incremental placement and lingual and facial curing. After the restorations were finished and polished, the margins of those in the rebonded groups were etched, rinsed, and dried. The adhesive resin, Single Bond, was applied at the composite resin-tooth interface and light-cured. All the specimens were thermocycled, stained with 1% methylene blue, sectioned, and evaluated for leakage (0-4 scale) by two examiners., Results: Almost no leakage occurred at enamel margins. At the cementum margins, differences in microleakage related to restorative material or technique were not statistically significant. However, leakage at the cementum margins was significantly greater than at the enamel margins for both composite resin materials.

This in vitro study evaluated the wear resistance of resin composite polymerized using four different light-curing systems. For this, a well-defined cylindrical cavity preparation (4.0 mm in diameter x 3.0 mm in depth) was made in a ceramic block (n=4 per material/light condition). Uncured material, either a universal hybrid composite (Herculite XRV) or a flowable hybrid composite (Revolution Formula 2), was packed and light-cured from the top surface only with one of the four light-curing units: 1) a conventional quartz-tungsten-halogen light, 2) a soft-start light, 3) an argon-ion laser or 4) a plasma-arc curing light. After storing the specimens in deionized water at 37 degrees C for 24 hours, the excess cured material was ground through successive grits up to a final 1200-grit SiC abrasive. The specimens were placed in deionized water at 37 degrees C for an additional 24 hours. Wear simulation was performed using a four-station Leinfelder-type three-body wear device. A slurry of water and unplasticized polymethylmethacrylate beads, simulating an artificial food bolus, was placed on the surface of each resin-composite-restored ceramic block. The entire cycling procedure was carried out 400,000 times. Impressions of each resin composite surface were taken with polyvinylsiloxane and epoxy replicas were made. Wear analyses were conducted by generating tracings across the worn surface of epoxy replicas using profilometer scans. For the universal hybrid composite and the flowable hybrid composite, the lowest wear occurred in specimens that were cured using the conventional quartz-tungsten-halogen light, and the highest wear was detected on those specimens made using the argon-ion laser. For both resin composites, the mean wear for specimens cured using the argon-ion laser was significantly higher than that of the specimens cured with the three other lights, which were statistically similar.

Ebola virus subtype Zaire (Ebo-Z) induces acute haemorrhagic fever and a 60-80% mortality rate in humans. Inflammatory responses were monitored in victims and survivors of Ebo-Z haemorrhagic fever during two recent outbreaks in Gabon. Survivors were characterized by a transient release in plasma of interleukin-1beta (IL-1beta), IL-6, tumour necrosis factor-alpha (TNFalpha), macrophage inflammatory protein-1alpha (MIP-1alpha) and MIP-1beta early in the disease, followed by circulation of IL-1 receptor antagonist (IL-1RA) and soluble receptors for TNFalpha (sTNF-R) and IL-6 (sIL-6R) towards the end of the symptomatic phase and after recovery. Fatal infection was associated with moderate levels of TNFalpha and IL-6, and high levels of IL-10, IL-1RA and sTNF-R, in the days before death, while IL-1beta was not detected and MIP-1alpha and MIP-1beta concentrations were similar to those of endemic controls. Simultaneous massive activation of monocytes/macrophages, the main target of Ebo-Z, was suggested in fatal infection by elevated neopterin levels. Thus, presence of IL-1beta and of elevated concentrations of IL-6 in plasma during the symptomatic phase can be used as markers of non-fatal infection, while release of IL-10 and of high levels of neopterin and IL-1RA in plasma as soon as a few days after the disease onset is indicative of a fatal outcome. In conclusion, recovery from Ebo-Z infection is associated with early and well-regulated inflammatory responses, which may be crucial in controlling viral replication and inducing specific immunity. In contrast, defective inflammatory responses and massive monocyte/macrophage activation were associated with fatal outcome.

Purpose: The purposes of this retrospective case series study were to evaluate safety issues and determine participants' perceptions of a nightguard vital bleaching (NGVB) technique approximately 10 years post-treatment (average, 118 mo; range, 108-144 mo)., Materials and Methods: The study sample included 30 (79%) of 38 participants who had completed a previous NGVB study using a 10% carbamide peroxide solution (Proxigel or Gly-Oxide) in a custom tray for 6 weeks. Participants were asked whether there had been any change in the shade of their teeth post-treatment and, if so, to quantify the change on a verbal scale. In addition, 19 participants had gingival index and tooth vitality evaluated clinically, external cervical root anatomy evaluated radiographically, and enamel surface changes evaluated microscopically., Results: Thirty-five (92%) of the original 38 participants had successful lightening of their teeth. At approximately 10 years post-treatment (average, 118 mo; range, 108-144 mo), external cervical resorption was not diagnosed and gingival index and tooth vitality findings were considered within the normal expectations for the sample studied, suggesting minimal clinical post-NGVB side effects at approximately 10 years. Scanning electron microscopic observations did not reveal substantial differences between treated and nontreated surfaces. Color stability, as perceived by 43% of the participants, may last approximately 10 years (average, 118 mo; range, 108-144 mo) post-treatment., Clinical Significance: Nightguard vital bleaching using 10% carbamide peroxide is safe and effective, with no noticeable side effects, at approximately 10 years (average, 118 mo; range, 108-144 mo) post-treatment. Color stability, as perceived by 43% of the participants, may last 108 to 144 months post-whitening.

In order to shed light on the mechanisms of antifilarial protective immunity, we investigated the course of experimental loaiosis after vaccination in a nonhuman primate host, Mandrillus sphinx. Six vaccinated (V) mandrills received 50 irradiated L3 while six nonvaccinated (NV) received saline solution on days -60, -30 and -15. All animals were challenged with 100 intact L3 (day 0). Parasitological and immunological status were followed for 9 months. Vaccination delayed the appearance and mean peak of microfilaraemia. Five mandrills (Mf-) were never microfilaraemic (one V mandrill) or microfilaraemic on only one occasion (2 V and 2 NV), the other seven having stable microfilaraemia (Mf+). The cytokine response of peripheral blood mononuclear cells to L3 (L3 Ag) was Th2 dominated, while microfilariae (Mf Ag) elicited a Th0-like response. During vaccination, Th2 cytokine production significantly increased in V mandrills against L3 Ag, as well as Mf Ag, whereas Th1 cytokines decreased. On day 60 postinoculation, cellular proliferation was higher in V mandrills in response to L3 and Mf Ags and PHA-L mitogen. At the end of prepatency (on day 130), mandrills with delayed appearance of microfilaraemia exhibited a high, transient IL-2 and IL-4 secretion in response to L3 Ag. Finally, high anti-Mf Th2 cytokine levels characterized Mf-mandrills not only during prepatency, but also (for IL-5) before immunization. However, the presence of a balanced Th1 anti-L3 response during prepatency in the amicrofilaraemic mandrill suggests its importance in protective immunity. Taken together, our data suggest that Th2 cells and also Th1 components of the antifilarial response, especially to larval antigen, may contribute to parasite elimination.

This study reports the first field evaluation of a new diagnostic technique for Ebola virus disease with sensitivity and specificity. Ebola virus causes rare but fulminating outbreaks in Equatorial Africa. Rapid differentiation from other infections is critical for timely implementation of public health measures. Patients usually die before developing antibodies, necessitating rapid virus detection. A reverse transcriptase-polymerase chain reaction (RT-PCR) assay was developed, implemented and evaluated at Centre International de Recherches Médicales de Franceville (CIRMF) in Gabon, to detect Ebola viral RNA in peripheral blood mononuclear cells (PBMC). Twenty-six laboratory-confirmed patients during and 5 after the acute phase of Ebola haemorrhagic fever, 15 healthy controls and 20 febrile patients not infected with Ebola virus were studied. RT-PCR results were compared with ELISA antigen capture, and Ebola specific IgM and IgG antibody detection. Ebola virus RNA was amplified from 26/26 specimens from the acute phase, 3/5 during recovery, 0/20 febrile patients and 1/15 negative controls. Sensitivity of RT-PCR in identifying acute infection and early convalescence compared with antigen or IgM detection was 100% and 91% respectively, and specificity compared with antigen detection and IgM assay combined was 97%. Antigen capture detected only 83% of those identified by PCR, and IgM only 67%. Ebola virus RNA was detected in all 13 fatalities, only 5 of whom had IgM and none IgG. RT-PCR detected Ebola RNA in PBMC one to three weeks after disappearance of symptoms when antigen was undetectable. RT-PCR was the most sensitive method and able to detect virus from early acute disease throughout early recovery., (Copyright 2000 Wiley-Liss, Inc.)

Background: Cardiac surgery continues to be limited by an inability to achieve complete myocardial protection. This may result from the use of hypothermic cardioplegia. Interestingly, the subcellular changes of animal hibernation parallel the altered biology of induced hypothermic myocardial ischemia, but are well tolerated by hibernated mammalian myocardium. Evidence indicates this protection is mediated by activation of the delta opioid receptor, which elicits profound metabolic effects at the whole animal, organ, and cell level. In this study, we sought to determine if pentazocine, with agonist activity at the delta opioid receptor, could improve myocardial recovery following global ischemia over a wide range of temperatures., Methods: Isolated rabbit hearts received either standard cardioplegia or were pretreated with racemic, d or 1 isomer pentazocine. Hearts were then subjected to 2 hours at 34 degrees C, or 3.5 hours at 20 degrees C, or 4 hours at 10 degrees C of cardioplegic ischemia and reperfused. Functional recovery was compared to controls., Results: Isovolumic developed pressure, coronary flow, oxygen consumption, and ultrastructural preservation were enhanced with pentazocine delta opioid mediated protection, which appears to be additive to standard cardioplegia, even at low temperatures., Conclusions: Teleologically, delta opioid protection parallels animal hibernation, which occurs from 34 degrees down to 0 degrees C. The use of delta opioid receptor agonists may have important clinical implications for cardiac surgery and deserves further study.

The HLA class II typing of 167 unrelated Gabonese individuals from the Banzabi ethnic group was assessed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The most frequent alleles at each locus were DRB1*1501-3 (0.31), DQA1*0102 (0.50), DQB1*0602 (0.42) and DPB1*0402 (0.29). The estimation of the haplotype frequencies as well as the observation of the segregation of several haplotypes using additional HLA typing of relatives, revealed that the three-locus haplotype DRB1*1501-3-DQA1*0102-DQB1*0602 was found at the highest frequency (0.31) among these individuals. This haplotype is not typically African and has already been described in Caucasians, but its presence at high frequency is exclusive to populations originating from Central Africa, and can thus be designated as a particular genetic marker of these populations.

Ebola virus is very pathogenic in humans. It induces an acute hemorrhagic fever that leads to death in about 70% of patients. We compared the immune responses of patients who died from Ebola virus disease with those who survived during two large outbreaks in 1996 in Gabon. In survivors, early and increasing levels of IgG, directed mainly against the nucleoprotein and the 40-kDa viral protein, were followed by clearance of circulating viral antigen and activation of cytotoxic T cells, which was indicated by the upregulation of FasL, perforin, CD28 and gamma interferon mRNA in peripheral blood mononuclear cells. In contrast, fatal infection was characterized by impaired humoral responses, with absent specific IgG and barely detectable IgM. Early activation of T cells, indicated by mRNA patterns in peripheral blood mononuclear cells and considerable release of gamma interferon in plasma, was followed in the days preceding death by the disappearance of T cell-related mRNA (including CD3 and CD8). DNA fragmentation in blood leukocytes and release of 41/7 nuclear matrix protein in plasma indicated that massive intravascular apoptosis proceeded relentlessly during the last 5 days of life. Thus, events very early in Ebola virus infection determine the control of viral replication and recovery or catastrophic illness and death.

During the 2 years 1993 to 1995, an entomological survey was carried out in the savanna-forest area of Franceville, Gabon, investigating malaria transmission in one suburban district of Franceville (Akou) and in one rural village (Benguia). The biting rates of the Anopheles vectors were 10 times higher in the rural zone compared to the suburban zone. Anopheles funestus Giles was the predominant species in both zones followed by Anopheles gambiae s.l. Giles. The densities of Anopheles nili Theobald and Anopheles moucheti Evans were very low. In the suburban zone, transmission was maintained throughout the year by An. funestus and An. gambiae s.l., whereas in rural zones the secondary vectors An. nili and An. moucheti were also involved in transmission. Humans in a suburban setting received one infective bite per person every 4 days, whereas in the rural area the infective biting rate was 4 times higher. Considering each vector, the observed entomological inoculation rates (EIRs) were one infective bite per person every 6 and 17 days for An. funestus and An. gambiae s.l., respectively, at Akou. At Benguia, the EIRs were one infective bite per person every 2, 3, 6, and 19 days for the 4 An. funestus, An. gambiae s.l., An. nili, and An. moucheti, respectively. The predominance of An. funestus over An. gambiae s.l. and its high EIR make it the most important malaria vector in this region of Haut-Ogooué.

From the end of 1994 to the beginning of 1995, 49 patients with hemorrhagic symptoms were hospitalized in the Makokou General Hospital in northeastern Gabon. Yellow fever (YF) virus was first diagnosed in serum by use of polymerase chain reaction followed by blotting, and a vaccination campaign was immediately instituted. The epidemic, known as the fall 1994 epidemic, ended 6 weeks later. However, some aspects of this epidemic were atypical of YF infection, so a retrospective check for other etiologic agents was undertaken. Ebola (EBO) virus was found to be present concomitantly with YF virus in the epidemic. Two other epidemics (spring and fall 1996) occurred in the same province. GP and L genes of EBO virus isolates from all three epidemics were partially sequenced, which showed a difference of <0.1% in the base pairs. Sequencing also showed that all isolates were very similar to subtype Zaire EBO virus isolates from the Democratic Republic of the Congo.

Although the WHO declared global smallpox eradication in 1980, the Orthopoxvirus remains a source of concern for several reasons. Firstly, stocks of the smallpox virus have been preserved for experimental use (at least officially in the USA and Russia) so that an escaped isolate could lead to reemergence and spread of the disease worldwide. Secondly discontinuation of smallpox vaccination programs has led to dwindling acquired immunity in the world population thus raising the risk of epidemic extension of several Orthopoxvirus zoonoses (e.g., monkeypox). Thirdly stocks of camelpox virus which is very similar to Smallpox virus and was intended for biological warfare were discovered during the Gulf War in 1991 and pose a potentially serious threat. Finally official stocks of Smallpox virus could be stolen and used by bioterrorists. Thus reemergence of Orthopoxvirus including smallpox, monkeypox, cowpox, and camelpox is a real danger and contingency planning is needed to define prophylactic and therapeutic strategies to prevent and/or stop an epidemics. Adverse effects from earlier smallpox vaccine (vaccinia) in healthy people or immunocompromised people (congenital or acquired as in HIV infected patients) are absolute contraindications to smallpox vaccination at this time. Further research is needed to develop new vaccines (e.g., attenuated isolates of vaccinia) and effective treatment. This is the only valid reasons for postponing planned destruction of remaining Smallpox virus stocks.

Two species of macaques, including two Macaca fascicularis from the Philippines, two M. fascicularis from Mauritius, and one Macaca mulatta, were experimentally infected with blood stages of Plasmodium coatneyi and followed during their clinical, parasitological, biological, and immunological evolution. Plasma cytokine (TNF-alpha, IL-1beta, IL-6, IFN-gamma) production peaked for all monkeys 11 days after inoculation, concomitantly with peaks of parasitemia. Only the M. fascicularis from the Philippines survived the infection. The main features, which discriminated nonfatal from fatal cases, were the observation in M. fascicularis from the Philippines of a mean CD4+/CD8+ ratio below I and of their ability, as revealed by mitogenic stimulation of whole blood, to produce increasing amounts of IFN-gamma as infection evolved. The contribution of environmental and genetic factors, which may differentiate the three groups of monkeys and therefore explain fatal or nonfatal evolution of the infection among them, is discussed.

Human T-cell leukemia virus type 1 (HTLV-1) is a member of the Oncoretrovirinae family containing several viruses that have been associated with a low incidence of leukemia and sarcoma in mammals. Primates are susceptible to viruses of genera HTLV (humans) and STLV (other primates). The high degree of homology in genomic arrangement of HTLV and STLV is probably due to the existence of a common simian ancestor. Most infections are asymptomatic but a few cases exhibit blood diseases, e.g., T-lymphoma or T-cell leukemia, or neurologic disease, mainly, HTLV-1 associated myelopathy and tropical spastic paraparesia (HAM-TSP). The four major modes of viral transmission are vertical transmission from mother to child either in utero or, more commonly, during breastfeeding, sexual intercourse, blood transfusion, and intravenous drug use. Geographic distribution of HTLV-1 and its confinement to a few well-defined ecosystems have yet to be explained. Diagnosis is now easy and can reduce transmission by intravenous drugs use. Development of a vaccine seems possible given the low genetic variability of this virus.

The development of control strategies for loiasis is of crucial importance in endemic areas and depends heavily on the accurate identification of occult-infected individuals. A polymerase chain reaction (PCR) and nested polymerase chain reaction (nested PCR) were developed and based on sequences of the repeat 3 region (15r3) of the gene encoding a Loa loa 15-kD protein. The assays was performed on 20 blood samples from occult-infected subjects and 30 from field-collected amicrofilaremic individuals. The size of the initial PCR product was 396 basepairs (bp). When this initial amplification using primers 15r3(1) and 15r3(2) was carried out for 30 cycles, the PCR products from three of the 20 occult-infected and five of the 30 amicrofilaremic individuals were visualized after electrophoresis by staining the gel with ethidium bromide. Subsequent Southern blotting and hybridization with the specific probe revealed hybridization in 19 of 20 occult-infected and 23 of 30 amicrofilaremic samples but only after two days of exposure of the blot to the x-ray film. When the nested PCR was carried out (product size = 366 bp, primers 15r3(3) and 15r3(4)), 19 of 20 occult-infected and 23 of 30 amicrofilaremic samples that were positive by Southern hybridization of the initial PCR products were strongly positive by staining with ethidium bromide. Qualitative Southern blotting of the nested PCR products using the same probe previously described confirmed the ethidium bromide staining results after a very short exposure time of 4 hr. These results demonstrate that the nested PCR amplification product is specific and that its sensitivity in detecting occult loiasis is 95%. This approach has significant promise for the screening of large human populations for active loiasis without the requirement for blotting and hybridization of the PCR products.

Natural SIVmnd and STLVmnd infections of mandrills in a colony at the Centre International de Recherches Médicales de Franceville (CIRMF) in Gabon were investigated by genetic analysis to determine the extent of intracolony transmission. SIVmnd pol sequence analysis indicates that the six strains present in the colony belong to the SIVmnd lentivirus subgroup previously defined according to the only available prototype sequence (SIVmndGB1), which originated from the same colony. The intraanimal nucleotide diversity (1.1-3.1%) was similar in range to that reported in individuals infected by other HIV/SIVs. The interanimal diversity (0.5-4.3%) was not significantly different from that observed in each individual mandrill, indicating an epidemiological link among the SIVmnd isolates of distinct animals within the colony. Phylogenetic analysis of these isolates, together with seroepidemiological and behavior surveillance within the colony, indicates a predominant male-to-male transmission of SIVmnd that probably occurred during bouts of interanimal aggression. Moreover, our results suggest one case of vertical transmission of SIVmnd from a naturally infected founder female to one of her six offspring. The first genetic analysis of STLV isolates from mandrills is also reported here. Partial tax/rex sequences were used to evaluate the diversity between seven STLVmnd isolates and their phylogenetic relationships with other known strains of human and nonhuman primate T cell leukemia virus, types I and II (PTLV-I/II). They all belong to the PTLV-I subtype, but two genetically distinct STLVmnd groups were evidenced within the mandrill colony. The phylogenetic analyses of the STLVmnd isolates, together with seroepidemiological and behavior surveillance of the mandrills, indicate that intracolony transmissions of STLVmnd are also predominantly the result of male-to-male aggressive contacts.

Background: The protective effects of myocardial preconditioning may occur by way of multiple mechanisms, with G-protein-mediated protein kinase C (PKC) translocation as a final common pathway. In this study we investigate the pharmacologic induction of preconditioning, by PKC translocation, using PKC agonists/antagonists to reveal its effects on contractile function after myocardial ischemia., Methods: Langendorff-perfused rabbit hearts received: (1) control; (2) dimethyl sulfoxide (vehicle); (3) acetylcholine (0.55 mmol/L; PKC agonist); (4) 1,2-s,n-dioctanoylglycerol (DOG; 22 mmol/L; PKC agonist); (5) chelerythrine (0.8 mmol/L; PKC antagonist); or (6) DOG-chelerythrine followed by a 2-hour ischemic period, using modified St. Thomas cardioplegia and a 45-minute reperfusion period. The period of ischemia was chosen so as to allow for improvement by appropriate agonists. To observe metabolic changes, tissue nucleotides and nucleosides were measured. Membrane and cytosolic fractions of PKC were determined by an anti-PKC antibody directed against the PKC delta isozyme. Lactate levels and myocardial pH were measured., Results: The PKC agonists DOG and acetylcholine showed the greatest recovery of developed pressure (68% +/- 2%, 60% +/- 9%, respectively). Although pH, lactate, and nucleotide levels were similar between groups at all times, myocyte PKC translocation demonstrated 25% of PKC delta isoforms on cell membrane sites during baseline, which shifted to 67% delta 17% with unprotected ischemia. DOG mimicked this shift with 58% delta 12% of PKC delta isoforms on membranes, which was also blocked by chelerythrine to 35% +/- 7%., Conclusions: These data demonstrate that PKC translocation results in improved postischemic function, not by alteration of energetics or metabolism, and deserves further investigation.

Human filariasis due to Loa loa differs from other filariasis in that the majority of infected subjects are without circulating microfilariae (occult loiasis). In search for alternative diagnostic methods, which do not depend on circulating microfilariae or the (rather infrequent) eye-passage of adult worms, it was shown earlier that IgG4 antibodies directed against Loa loa adult worm antigen are apparently a good marker of occult loiasis and specific with regard to the sympatrically occurring Mansonella perstans. In this study we evaluated an IgG4 antibody-based ELISA using crude extract of Loa loa microfilariae (which is easier to obtain than adult worm) to estimate the prevalence of loiasis in 3 villages in South-East Gabon. Of 222 examined individuals (80 children < 16 years, 142 adults) 44 (20%) carried Loa loa microfilariae and 170 (77%) M. perstans. Using the mean OD-value + 1 standard deviation of 9 sera from patients solely infected with M. perstans (from the Gambia, where Loa loa is not endemic) as a cut-off, 35 of the 44 microfilaraemic Loa loa patients and 2 of the 9 Gambian controls were positive. This shows that our method had a sensitivity of 80% and a specificity of 78%. Among the remaining 178 subjects who had no microfilariae of Loa loa, as many as 97 (55%) had significant levels of specific IgG4 antibodies against Loa loa, suggesting that they carried occult loiasis. The mean IgG4 level in these putatively occult loiasis patients was slightly but significantly lower than in microfilaraemic subjects (P < 0.03). In conclusion, despite the limited sensitivity and specificity of our method, IgG4- ELISA at present is a very useful tool in estimating the real prevalence of loiasis in epidemiological surveys and at the individual level can confirm the diagnosis of L. loa amicrofilaraemic subjects with clinical signs suggesting loiasis.

The in vitro activities of new organometallic chloroquine analogs, based on 4-amino-quinoleine compounds bound to a molecule of ferrocene, were evaluated against chloroquine-susceptible, chloroquine-intermediate, and chloroquine-resistant, culture-adapted Plasmodium falciparum lineages by a proliferation test. One of the ferrocene analogs totally restored the activity of chloroquine against chloroquine-resistant parasites. This compound, associated with tartaric acid for better solubility, was highly effective. The role of the ferrocene in reversing chloroquine resistance is discussed, as is its potential use for human therapy.

Using the cluster-sampling method, the authors estimated the seroprevalence of 4 sexually transmitted diseases (STDs) among the sexually active general population in a city of 30,000 inhabitants in the east of Gabon. The seroprevalences were 2% for HIV-1, 13.8% for hepatitis B, 8.6% for Treponema pallidum and 59.6% for Chlamydia trachomatis. The seroprevalences of hepatitis B and chlamydia were stable over time and similar to those registered in other countries of central Africa. On the other hand, the seroprevalence of T. pallidum is notably low in comparison with these countries and seems to be decreasing. The seroprevalence of HIV-1 is also low but has doubled in 8 years in the city. Immigrant women from west Africa were a high-risk group for STDs but more generally, cohabiting was a risk factor for women.

The Ebola virus is an RNA virus of Filoviridae family. The earliest documented fatal epidemic of Ebola hemorrhagic occurred in 1976. There are four genetically different subtypes of Ebola virus. The virus remains in the blood for several weeks, can maintain its infectivity for several weeks at 20 degrees C outside the body, and survives for several weeks in corpses. Isolation of Ebola virus requires level 4 laboratory security conditions. Specimens are obtained by culturing mammal cells. Identification is achieved using reference serums. Serologic diagnosis is made using mainly ELISA technique for immunocapture of IgM or EBO Ag. The natural reservoir for Ebola virus is unknown. One possibility is that each isolated strain has a different reservoir. In recorded outbreaks, the index case has often had a history of contact with non-human primates. However since these animals are also highly sensitive to the virus, they cannot be considered as reservoirs but only as intermediate hosts. Transmission requires close contact such as occurs in association with health care, local customs, or funeral rites. In humans, infection causes hemorrhagic fever that progresses to diarrhea within 5 to 10 days. Recovery is observed in only 25% of cases. During outbreaks containment depends on implementation of simple precautions including isolation of suspected cases, appropriate protective clothing, disinfection with hypochlorite solutions, and proper waste disposal.

We investigated the early interactions between HIV-1, HIV-2, and simian immunodeficiency virus (SIV) envelope glycoproteins gp120(IIIB), gp105(ROD), and gp120(mac251), and human and macaque cells of the lymphocytic series. Our results demonstrate that the soluble viral glycoproteins induce a specific phospholipase A2 (PLA2) activation in lymphocytes through CD4. This PLA2 activation was induced after envelope glycoprotein-CD4 interaction and, because of its local membrane-destabilizing effect, may have important implications for preparing the lymphocyte membrane for fusion with the viral particle. However, this effect is not sufficient to accomplish fusion. These data indicate that the specific step of fusion may be downstream from PLA2 activation.

The efficacy of a 3-day clindamycin-quinine regimen to treat clinical malaria attacks was investigated in 256 children from western Gabon. Treatment was well tolerated by all of the children and its efficacy was higher than 97% by day 20. Thus this 3-day clindamycin-quinine regimen might constitute a potential alternative to chloroquine for treating clinical malarial attacks in children from Gabon.

Accurate and specific diagnosis of human loiasis is of crucial importance in an endemic area where two-thirds of infected individuals are without circulating microfilariae (occult loiasis). By using the polymerase chain reaction (PCR) and specific primers to the repeat 3 region (15r3) of the gene coding for Loa loa 15-kDa polyprotein antigen, DNA was amplified from total blood lysate of occult-infected subjects. A 396-bp DNA fragment was specifically detected. We tested the specificity of this method by qualitative hybridization to PCR products using blood lysates of the following subjects: (1) from Gabon (80 individuals residing in L. loa endemic area where loiasis exists sympatrically with Mansonella perstans); (2) from Togo (12 individuals infected with Onchocerca volvulus and M. perstans); (3) from Tahiti (12 individuals infected with Wuchereria bancrofti); and (4) from Mali (12 individuals infected with O. volvulus and M. perstans). Samples from Gabon included 60 L. loa amicrofilaremics and 20 L. loa occult-infected subjects. Qualitative hybridization carried out at 50 degrees C on PCR products, using a 15r3-specific oligonucleotide probe, revealed hybridization with L. loa-infected samples from Gabon and four samples from Togo after 2 days exposure to the film. The positive samples from Togo were characterized by the use of nested PCR. Three nested PCR products have been sequenced. No differences were observed between the three sequences and they are 99.72% identical to L. loa 15r3. None of bancroftian-infected individuals from Tahiti, nor O. volvulus- and M. perstans-infected individuals from Mali reacted after 1 week's exposure (overexposure) to the film. This allows us to conclude first that our 15r3 PCR assay is specific for L. loa and secondly that L. loa infections occur in Togo. The sensitivity of this 15r3 PCR assay was further investigated with occult patients and field-collected amicrofilaremic samples. We found that 19 of the 20 occult-infected individuals were positive on Southern hybridization, whereas 35/60 amicrofilaremics were positive. These results have shown that the sensitivity of this assay in detecting unequivocal, parasitologically proven occult loiasis was 95%, while the specificity with regard to the sympatric M. perstans was 100%.

Human infection with the parasite Loa loa is characterized by a good adaptation between the parasite and its host. One portion of the human population harbors only adult worms in subcutaneous tissues, whereas another portion also harbors the L1 microfilarial stage in peripheral circulation. This study was undertaken to understand the mechanisms by which the parasite evades or modulates host immunological attack. The cellular responses, based on T-cell proliferation, to the production of various cytokines (interleukin-2 [IL-2], gamma interferon [IFN-gamma], IL-4, and IL-5) and to expression of cytokine (IL-2, IFN-gamma, IL-4, IL-5, IL-10, and IL-12) mRNAs were investigated during the experimental infection with human parasite L. loa of a nonhuman primate which has been shown to display a spectrum of disease similar to that found in humans. Our results indicate that a T-cell unresponsiveness occurs when female worm products are released into the peripheral circulation, preceded by a transient period of strong T-cell proliferation, cytokine production, and cytokine mRNA expression. In the unresponsive state, only IL-10 mRNA is expressed, suggesting a role for IL-10 in down-regulation and maintenance of unresponsiveness. Taken together, these results indicate that both IL-10 production, which is known to inhibit B7 expression on monocytes, and the massive release of female products in the blood where T cells encounter antigens presented by nonactivated B lymphocytes, which lack costimulatory signals, should contribute to the inactivation of T cells.

The proliferation and cytokine profiles of peripheral blood mononuclear cells (PBMC) from microfilaraemic (Mf+) subjects infected by Loa loa in response to antigens of several parasitic stages were compared with those from amicrofilaraemic (Mf-) individuals. While a strong lymphoproliferative response and consistent levels of both Th1 (IL-2, interferon-gamma (IFN-gamma)) and Th2 (IL-4, IL-5) type cytokines were observed in response to adult worm (AW) and microfilariae (Mf) antigen in Mf- individuals, Mf+ subjects were characterized by a T cell unresponsiveness, including proliferation, cytokine production and IL-2 mRNA expression. Conversely, T cell responsiveness to mitogens and non-specific antigen were similar in the two endemic populations. Depletion of lymphocyte subpopulations indicated that T CD4+ were mainly involved in the specific cellular response. In contrast to other cytokines, IL-10 was produced in response to all parasitic stages, in both Mf+ and Mf- patients. Neutralization of IL-10 did not restore cytokine production in Mf+ patients, while B7 mRNA expression was similar between Mf+ and Mf- subjects in response to Mf antigen, suggesting that IL-10 was not the only factor responsible for T cell unresponsiveness. Mf+ patients have lower Mf antigen-specific IgG levels compared with Mf-, and there is a significant correlation between Mf antigen-specific antibodies and IL-5 responses. These findings suggest that Mf- status is correlated with T helper responsiveness, including proliferation and production of both Th1- and Th2-type cytokines, whereas Mf+ status is characterized by unresponsiveness of the same cell population, induced and/or maintained by microfilariae.

The International Center for Medical Research (CIRMF) is located in an area highly endemic for malaria in southeastern Gabon, where humans and apes (gorillas and chimpanzees) are living in the same geographic area. The presence of the CIRMF primate center housing apes (59 chimpanzees and nine gorillas in 1994) within the city of Franceville provided an opportunity to investigate the capability of cross-transmission of malaria species from humans to apes. The main vector of human malaria, Anopheles gambiae, was found in the primate center and in a nearby populated area of Franceville. Despite high malaria transmission in humans of both Plasmodium falciparum and P. malariae (mean of 43% cumulative prevalence in schoolchildren), none of the apes were found infected with plasmodia during a six-month investigation. However, low antibody levels against sporozoite and blood stages of both P. falciparum and P. malariae were detected in a few chimpanzees and gorillas. These results demonstrate that only rarely would apes be bitten in the field by mosquitoes infected with human malaria parasites. In the case of infection proven by serology, we did not find any evidence that blood-stage malaria parasites were able to the gametocyte stage. The absence of any established malaria transmission cycle within the primate colony of CIRMF indicates that apes cannot be considered as animal reservoirs for human malaria parasites in this environment.