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We have previously reported a strategy for production in Escherichia coli of recombinant immunogens fused to a hydrophobic tag to improve their capacity to associate with an adjuvant formulation [Andersson et al., J. Immunol. Methods 222 (1999) 171]. Here, we describe a further development of the previous strategy and present significant improvements. In the novel system, the target immunogen is produced with an N-terminal affinity tag suitable for affinity purification, and a C-terminal hydrophobic tag, which should enable association through hydrophobic interactions of the immunogen with an adjuvant system, here being immunostimulating complexes (iscoms). Two different hydrophobic tags were evaluated: (i) a tag denoted M, derived from the membrane-spanning region of Staphylococcus aureus protein A (SpA), and (ii) a tag denoted MI consisting of the transmembrane region of hemagglutinin from influenza A virus. Furthermore, two alternative affinity tags were evaluated; the serum albumin-binding protein ABP, derived from streptococcal protein G, and the divalent IgG-binding ZZ-domains derived from SpA. A malaria peptide M5, derived from the central repeat region of the Plasmodium falciparum blood-stage antigen Pf155/RESA, served as model immunogen in this study. Four different fusion proteins, ABP-M5-M, ABP-M5-MI, ZZ-M5-M and ZZ-M5-MI, were thus produced, affinity purified and evaluated in iscom-incorporation experiments. All of the fusion proteins were found in the iscom fractions in analytical ultracentrifugation, indicating iscom incorporation. This was further supported by electron microscopy analysis showing that iscoms were formed. In addition, these iscom preparations were demonstrated to induce M5-specific antibody responses upon immunisation of mice, confirming the successful incorporation into iscoms. The novel system for hydrophobic tagging of immunogens, with optional affinity and hydrophobic tags, gave expression levels that were increased ten to fifty-fold, as compared to the earlier reported system. We believe that the presented strategy would be a convenient way to achieve efficient adjuvant association for recombinant immunogens.
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PMID:Improved systems for hydrophobic tagging of recombinant immunogens for efficient iscom incorporation. 1075 48

Baculovirus is a viral pathogen of insects in general and lepidoptera in particular. The genome of this large virus consists of a circular, infectious bicatenary DNA molecule. At the end of its replication cycle in insects, baculovirus produces a large quantity of at least two proteins, i.e., polyedrine and polypeptide P10. These proteins are essential for transmission of the virus in nature, but are not necessary in cell cultures. Using molecular recombinant techniques, one or both of the genes coding for these proteins can be replaced by heterologous genes. In this way, baculovirus raised in vitro can be used to produce large quantities of the alien proteins at the end of the multiplication cycle. So far more than 3,000 different proteins have been expressed including several presenting interest as diagnostic tools (Puumala virus, Herpes simplex virus) or therapeutic treatment in man and animals (vaccinations against dengue, flu, malaria and production of anti-Rhesus immunoglobulins). Since it is based on the use of lepidoptera virus, this system would appear to be particularly safe. No vertebrate virus is able to replicate in the cell system used. Use of this genetic engineering tool will undoubtedly expand and holds great promise for the future.
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PMID:[Baculovirus: an example of an insect virus of use to humans]. 1090 57

Plasmodium falciparum infection in humans leads to a variety of symptoms ranging from an influenza-like syndrome to life-threatening complications. Animal models are useful tools for the detailed analysis of the interaction between both parasite and host factors leading to these various clinical manifestations. In this review, examining the different clinical, parasitological and haematological parameters associated with P. falciparum infection in spleen-intact monkeys, we propose this model as a good alternative for exploring some aspects of the host-parasite relationship in malaria.
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PMID:Plasmodium falciparumin the squirrel monkey (Saimiri sciureus): infection of non-splenectomised animals as a model for exploring clinical manifestations of malaria. 1096 78

It is extremely important to look for tropical and other exotic diseases in travellers who return with illness or become ill after travelling. Especially tropical diseases and exotic infectious diseases have to be excluded because of their possible fatal outcome. On the other hand, many travellers return with 'common' not-exotic illnesses not related to their journey. When in such cases attention is only given to exotic causes of their illness, diagnosis can be delayed which may be harmful. This was the case in 5 patients: a woman aged 44 years who suffered for months from bloody diarrhoea since her return from Brasil, due to a rectal adenocarcinoma, a 61-year-old man with diarrhoea upon returning from Egypt, who had hairy-cell leukaemia, a 17-year-old boy who developed a ketoacidotic diabetic crisis whilst on a journey in Uganda, but in whose case the first thoughts went to malaria, a 50-year-old man who suffered from throat pain since a journey through East Africa, during which he contracted a flu-like disease, and in whom Kahler's disease was diagnosed, and 69-year-old man suffering from recurrent fever and cough, in whom a radiological lesion was observed in the thorax which proved to be part of Wegener's disease.
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PMID:[Illness after travel not always due to exotic disease]. 1123 88

CD8+ T cells have been implicated as critical effector cells in protection against the pre-erythrocytic stage of malaria in mice and humans following irradiated sporozoite immunization. Immunization experiments in animal models by several investigators have suggested different strategies for vaccination against malaria and many of the targets from liver stage malaria antigens have been shown to be immunogenic and to protect mice from the sporozoite challenge. Several prime/boost protocols with replicating vectors, such as vaccinia/influenza, with non-replicating vectors, such as recombinant particles derived from yeast transposon (Ty-particles) and modified vaccinia virus Ankara, and DNA, significantly enhanced CD8+ T cell immunogenicity and also the protective efficacy against the circumsporosoite protein of Plasmodium berghei and P. yeti. Based on these experimental results the development of a CD8+ T cell inducing vaccine has moved forward from epitope identification to planning stages of safety and immunogenicity trials of candidate vaccines.
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PMID:Protective CD8+ T cell responses against the pre-erythrocytic stages of malaria parasites: an overview. 1128

We reviewed 232 consecutive patients admitted to a tertiary-care hospital under the care of an infectious diseases unit for management of febrile illness acquired overseas. A total of 53% presented to hospital within 1 week of return and 96% within 6 months. Malaria was the most common diagnosis (27% of patients), followed by respiratory tract infection (24%), gastroenteritis (14%), dengue fever (8%), and bacterial pneumonia (6%). Pretravel vaccination may have prevented a number of admissions, including influenza (n=11), typhoid fever (n=8) and hepatitis A (n=6). Compared to those who had not traveled to Africa, those who had were 6 times more likely to present with falciparum than nonfalciparum malaria. An itinerary that included Asia was associated with a 13-fold increased risk of dengue, but a lower risk of malaria. Palpable splenomegaly was associated with an 8-fold risk of malaria and hepatomegaly with a 4-fold risk of malaria. As a cause of fever, bacterial pneumonia was > or =5 times more likely in those who were aged >40 years.
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PMID:Fever in returned travelers: review of hospital admissions for a 3-year period. 1148 83

The aim of this article is to highlight the importance of the history of public health for public health research and practice itself. After summarily reviewing the current great vitality of the history of collective health oriented initiatives, we explain three particular features of the historical vantage point in public health, namely the importance of the context, the relevance of a diachronic attitude and the critical perspective. In order to illustrate those three topics, we bring up examples taken from three centuries of fight against malaria, the so called "re-emerging diseases" and the 1918 influenza epidemic. The historical approach enriches our critical perception of the social effects of initiatives undertaken in the name of public health, shows the shortcomings of public health interventions based on single factors and asks for a wider time scope in the assessment of current problems. The use of a historical perspective to examine the plurality of determinants in any particular health condition will help to solve the longlasting debate on the primacy of individual versus population factors, which has been particularly intense in recent times.
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PMID:History of health, a valuable tool in public health. 1151 33

Malaria was once one of the most common diseases in Uzbekistan. There were massive epidemics with high mortality rates, wherein 140,000 to 700,000 cases of malaria were recorded. Following large-scale malaria control measures, the disease was eradicated in Uzbekistan in 1961 and the epidemiological situation is still favorable. The natural and climatic conditions that prevail in the Republic of Uzbekistan mean that the country is very susceptible to malaria. There are large water areas varying in type and origin, which provide a habitat for a number of epidemiologically dangerous species of malaria-transmitting mosquitoes in a single area. These are Anopheles maculipennis, An. pulcherrimus and An. superpictus. The prevailing temperatures promote rapid growth of vector mosquitoes and parasites and the malaria transmission season is over 5 months long. Seven malaria-transmitting mosquito species have been recently recorded in the Republic. DDT resistance has been so far noted in Anopheles maculipennis, An. hyrcanus and An. bifurcatus. An. superpictus is sensitive to all insecticides used in clinical practice (organophosphorus and organochlorine compounds, HOS, carbamates, pyrethroids). The most dangerous areas for transmitting malaria by importation are the flood plains of the country's main rivers, such as Syrdarya, Amudarya, Chirchik, Surkhana, etc., and rice-growing areas (an area of about 150,000 ha was under rice cultivation in 1999). The Republic is still very subjected to large-scale importations of malaria particularly in the towns and areas along the border with Tajikistan. There has been recently an increase in the incidence of infections imported into the Republic: 27 cases in 1995, 51 in 1996, 52 in 1997, 74 in 1998, and 78 in 1999. Eight regions of Uzbekistan border Tajikistan, their population is over 5.6 million people. In addition, close family ties between the populations of the frontier towns and regions further increase the risk for malaria to be imported and passed on. Noteworthy is the Surkhandaryin region that accounted for 60% of the cases recorded in 1999. The number of towns and villages where malaria occurs for the first time increased (49 and 46 cases in 1999 and 1998, respectively). The number of cases imported into rural areas also increased (70 (83%) cases in 1999 versus 48 (65%) cases in 1998); due to the large populations of malaria mosquitoes, there is a real danger that the disease may spread. In 1999, most cases of malaria were imported from Tajikistan (65 cases or 76% of all cases). There was a case from each of the following countries: Afghanistan, Pakistan, and Kazakhstan and 5 cases from Azerbaijan and Kyrgyzstan. The recorded cases included slighly more men than women (54% vs 46%). There were 10 infected children under 14 years, which was 23.5% of all notified cases. Analyzing various populations showed that 67.1% of the patients visited their relatives in malaria-endemic countries (mostly Tajikistan) and 25.8% migrated from Tajikistan. All the detected cases were confirmed by laboratory tests. As in the past, most cases were tertian (P. vivax) malaria (n = 82 or 96.4% of all cases). Tropical (P. falciparum) malaria was confirmed in 3 (3.5%) cases. These cases had been imported from Tajikistan into the Surkhandaryin region. Seventy seven (91%) cases were detected in the epidemical season. Of them 58 (68.2%) cases were detected during a malaria transmission season. Seven cases who contacted the patients with imported malaria and were infected were recorded in 1999. They included 4 and 3 cases in the Surkhandaryin and Kashkadaryin Regions, respectively. In 1999, there was a decline in the number of malaria patients who needed health care and in the diagnosed malaria cases in therapeutical and prophylactic institutions. Throughout the country, 34 (40%) of the 85 detected cases presented within 3 days of malaria outbreak (68.9% in 1998). Malaria was immediate diagnosed in 43.5% of cases (64.9% in 1998). The remaining cases were diagnosed as having acute respiratory viral infections, tropical and parasitic diseases, viral hepatitis, or influenza. Early diagnosis of malaria was made in 60% of cases (77% in 1998). Three cases of imported tertian malaria were recorded in the Tashkent Region in the first quarter of 2000. They were imported from Tajikistan into rural areas and the patients had been infected during the 1999 season. Epidemiological surveillance of malaria in Uzbekistan is regularly carried out by the general network of health facilities and by the departments of parasitology of state epidemiological surveillance centers in collaboration with medical administrative departments, the Ministry of Agriculture and Fisheries, the L.M. Isayev Research Institute of Medical Parasitology, and other agencies. Active links are maintained with WHO under the Roll Back Malaria programme. Great emphasis is laid on medical staff training at all levels. During the 1999 epidemiological survey, 672,536 laboratory tests were performed on blood samples from suspected malaria patients and individuals who had visited malaria-endemic countries, 55% of them suffering from fever. A total area of 17 million m2 of dwelling and nondwelling buildings 20 ha of water areas were treated against mosquitoes and the larvivorous fish Gambusia was put into the water areas occupying 6,500 ha. In all cases of malaria, the focus of infection was epidemiologically surveyed and required epidemic preventive measures were implemented. All malaria patients received a full course of radical therapy and recovered completely. The epidemiological surveillance system for malaria is affected by staff shortages at the parasitology departments of state epidemiological surveillance centers and by shortages of microscopes, reagents, sterilizing equipment, insecticides, etc. There are still difficulties in obtaining supplies of primaquine although a small stock is locally available as due to WHO humanitarian assistance. The Epidemiological Malaria Surveillance Programme for the Republic of Uzbekistan for 2000-2004, intended to strengthen the epidemic control capacity of health care facilities, Ministry of Health, is under adoption. The following activities are scheduled for 2000: to plan malaria control activities, including the zoning of the country by the risk of malaria transmission in accordance with republic-leveled directives, instructions, and methodology and WHO recommendations: adjustments to these plans to be made as necessary; to fill vacant posts in the parasitology departments of state epidemiological surveillance centers; to procure stocks of antimalarial drugs, reagents, insecticides, sterilizing equipment, etc., to be paid for from epidemiological service resources; to include malaria issues into certifying tests for physicians, as appropriate for the posts to be occupied and their level of qualifications; to publish posters, brochures, and leaflets about malaria prevention before the malaria transmission season for health education; to hold seminars and meetings for health workers on the etiology of malaria, its clinical features, diagnosis, treatment, and prevention.
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PMID:[Current malaria situation in the Republic of Uzbekistan]. 1154 13

Camillo Golgi opened new avenues in histology and neurobiology as well as in clinical neurology and internal medicine. This is demonstrated by investigations Golgi performed during 1861-1876 on the etiology and pathology of mental diseases, on the neuropathology of Huntington's chorea (Golgi provided the first detailed description at the microscopic level of pathological changes in the basal ganglia and cerebral cortex of one case of chorea), on meningiomas and cerebral gliomas. In the period following these investigations, Golgi focused especially on infectious diseases. He pursued fundamental studies on malaria (which remain among the most important and original of his contributions) on rabies, as well as on smallpox and influenza. Thus, Camillo Golgi should be remembered for his discoveries of the black reaction and the Golgi apparatus, as well as for the modern impulse he was able to give to clinical neurology and internal medicine.
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PMID:Camillo Golgi: a clinical pathologist. 1162 95

Research efforts to develop DNA vaccine products have progressed to the clinical evaluation of several vaccine candidates. These clinical trials are assessing safety and immunogenicity and whether different routes and schedules of administration can improve immune responses. Vaccine candidates under study include those for hepatitis B, HSV-2, HIV-1, influenza, and malaria. Unique safety concerns include the potential for genomic integration, biodistribution, tolerance and auto-immunity. CBER has issued a Points to Consider (PTC) document outlining regulatory policy, recommendations for the development of consistent manufacturing processes, and the design of preclinical studies to establish the safety of DNA vaccine products. This presentation will provide an overview of DNA vaccine products currently under clinical evaluation and the regulatory policy that has guided their development.
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PMID:Regulation and review of DNA vaccine products. 1171 25


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