Observation of structural anisotropy and the onset of liquidlike motion during the nonthermal melting of InSb.

The melting dynamics of laser excited InSb have been studied with femtosecond x-ray diffraction. These measurements observe the delayed onset of diffusive atomic motion, signaling the appearance of liquidlike dynamics. They also demonstrate that the root-mean-squared displacement in the [111] direction increases faster than in the [110] direction after the first 500 fs. This structural anisotropy indicates that the initially generated fluid differs significantly from the equilibrium liquid.

Clocking femtosecond X rays.

Linear-accelerator-based sources will revolutionize ultrafast x-ray science due to their unprecedented brightness and short pulse duration. However, time-resolved studies at the resolution of the x-ray pulse duration are hampered by the inability to precisely synchronize an external laser to the accelerator. At the Sub-Picosecond Pulse Source at the Stanford Linear-Accelerator Center we solved this problem by measuring the arrival time of each high energy electron bunch with electro-optic sampling. This measurement indirectly determined the arrival time of each x-ray pulse relative to an external pump laser pulse with a time resolution of better than 60 fs rms.

Atomic-scale visualization of inertial dynamics.

The motion of atoms on interatomic potential energy surfaces is fundamental to the dynamics of liquids and solids. An accelerator-based source of femtosecond x-ray pulses allowed us to follow directly atomic displacements on an optically modified energy landscape, leading eventually to the transition from crystalline solid to disordered liquid. We show that, to first order in time, the dynamics are inertial, and we place constraints on the shape and curvature of the transition-state potential energy surface. Our measurements point toward analogies between this nonequilibrium phase transition and the short-time dynamics intrinsic to equilibrium liquids.

Diffraction imaging of single particles and biomolecules.

Theory predicts that with a very short and very intense X-ray pulse, the image of a single diffraction pattern may be recorded from a large macromolecule, a virus, or a nanocluster of proteins without the need for a crystal. A three-dimensional data set can be assembled from such images when many copies of the molecule are exposed to the beam one by one in random orientations. We outline a method for structure reconstruction from such a data set in which no independent information is available about the orientation of the images. The basic requirement for reconstruction and/or signal averaging is the ability to tell whether two noisy diffraction patterns represent the same view of the sample or two different views. With this knowledge, averaging techniques can be used to enhance the signal and extend the resolution in a redundant data set. Based on statistical properties of the diffraction pattern, we present an analytical solution to the classification problem. The solution connects the number of incident X-ray photons with the particle size and the achievable resolution. The results are surprising in that they show that classification can be done with less than one photon per pixel in the limiting resolution shell, assuming Poisson-type photon noise in the image. The results can also be used to provide criteria for improvements in other image classification procedures, e.g., those used in electron tomography or diffraction.

Visceral leishmaniasis in Somalia. Significance of IgG subclasses and of IgE response.

We have determined the levels of IgG subclasses and IgE as well as specific antibodies of these isotypes in sera from 22 patients with clinical visceral leishmaniasis (VL) from Somalia. The results are compared with those obtained from 30 Somali and 23 Swedish controls. We found markedly increased concentrations of IgG1 in the VL sera, indicating that the pronounced increase in IgG in VL which is generally considered to be due to polyclonal B-cell activation is mainly restricted to this subclass. The IgG2 concentrations were significantly decreased. The IgG3 and IgG4 concentrations, on the other hand, did not differ between the two groups of Somali sera. The Somali control sera contained higher concentrations of IgG1 and IgG3, but significantly lower concentrations of IgG2 as compared to Swedish controls. The IgG4 values, on the other hand, were not different between the two groups of control sera. Anti-leishmania antibodies belonging to all IgG subclasses, were found in the patients' sera. There was no significant difference in total IgE between sera from VL patients and controls and specific IgE antibodies were only detected in a few patients. The Western blot assay (WB), revealed the presence of two bands corresponding to 74 kDa and 88 kDa in all patients' sera, indicating a possible diagnostic role for WB in this particular population.

Visceral leishmaniasis in Somalia: prevalence of markers of infection and disease manifestations in a village in an endemic area.

Prevalence and disease manifestations of visceral leishmaniasis (VL) were studied in a Somali village in an area which has long been known to be endemic for VL. Demographic data were collected from 102 households, comprising 438 inhabitants. Clinical examination was performed of 306 individuals, 72% of the 426 eligible persons. Of these, 276 (90%) agreed to give blood and 246 (80%) to be skin tested with leishmanin. Leishmanin reactions were positive; in 26% anti-Leishmania antibodies were detected in 11%, and splenomegaly was recorded in 14% (23% of those who were seropositive). Malaria was hypoendemic and therefore unlikely to be responsible for more than 10% of the cases with splenomegaly. Three of the seropositive villagers with splenomegaly complained of feeling ill. The remaining 91 sero- and/or leishmanin-positive individuals had no complaint regarding their health and had not experienced any long period of illness. There was a slight over-representation of males in the group of sero- and/or leishmanin-positive villagers, possibly due to a gender-associated difference in exposure to the parasite. Among the patients with clinical VL treated at Mogadishu hospitals during 1989 and 1990, the male/female ratio was 3.3:1, which may indicate a selection of male patients for hospital care. Most patients were < or = 15 years old, suggesting that the highest risk of becoming clinically ill was among children.

An epidemic of Plasmodium falciparum malaria in Balcad, Somalia, and its causation.

The causative factors of an epidemic of falciparum malaria were investigated in Balcad, Somalia, a town with previously low malaria transmission, where malaria incidence rose more than twenty-fold between 1986 and 1988. The emergence of chloroquine resistance, accelerated by high drug pressure, low herd immunity and favourable meteorological conditions were identified as major causes of the epidemic. Chloroquine resistance of grades RII and RIII was first observed in Balcad in 1987 and rapidly increased to 72% of the Plasmodium falciparum infections in 1988. In the absence of alternative treatment, resistance resulted in the accumulation of a massive infective reservoir and therefore increased malaria transmission, associated with intensive clinical symptomatology. The advent of chloroquine resistance was less violent in the area of Malable, where malaria is stable and communal immunity higher than in Balcad.

Visceral leishmaniasis in Somalia: prevalence of leishmanin-positive and seropositive inhabitants in an endemic area.

In an endemic area of Somalia both humoral and cell mediated immunity against Leishmania donovani was demonstrated in 246 inhabitants. In a study of 14 patients with active visceral leishmaniasis, we found that antibodies appear early in infection and that they are then demonstrable for a limited period only. Leishmanin positivity develops later and persists longer, but does not seem to be lifelong. The majority of the immunoreactive individuals were either sero- or leishmanin positive. This finding is in accord with the result obtained in recent experimental studies indicating a regulatory effect exerted on humoral and cell mediated immunity by different T lymphocyte subsets.

Visceral leishmaniasis in Somalia. Circulating antibodies as measured by DAT, immunofluorescence and ELISA.

Sera from patients with visceral leishmaniasis (VL) (n = 26), healthy residents of Mogadishu (n = 157), inhabitants of a village in an endemic area (n = 276) and healthy Swedes (n = 60) were examined using the direct agglutination test (DAT), immunofluorescence (IF) and ELISA for antibodies against Leishmania donovani. The study was carried out in order to provide baseline data for antibody responses in visceral leishmaniasis as existing in Somalia and to explore which one of these methods would be most suitable for diagnosis of clinical cases as well as for epidemiological population studies in Somalia. All patients had high levels of circulating antibodies, however, lower values were recorded in the early stages of the disease. High reactivity in ELISA was seen first after one year. All three tests distinguished well between sera from VL patients and healthy controls. Approximately 10% of the sera from villagers were reactive above the cut-off levels in the three tests. DAT is the simplest to perform and does not require much equipment. ELISA can be made simple and economic if performed in one serum dilution and read visually. IF requires more expensive and specialized equipment and is not suitable for large scale examination of sera. A complete evaluation of the three tests should also include the analysis of sera from various stages and manifestations of the disease.

Reference ranges for IgG, IgM and IgA in the serum of urban and rural Somalis.

In order to provide baseline data for an immuno-parasitological laboratory in Somalia, serum concentrations of IgG, IgM and IgA were determined in some key populations: healthy residents of Mogadishu (n = 157), inhabitants of the village of Daimo Samo (n = 276) and patients with malaria (n = 39) and visceral leishmaniasis (n = 26), both protozoan infections accompanied by hypergammaglobulinaemia and causing severe health problems in Somalia. Since the serum immunoglobulin concentrations in the Somali populations studied were not normally distributed, they were evaluated using medians and percentiles. Significantly higher values of IgG, IgM and IgA were demonstrated in healthy Mogadishu residents as compared to healthy Swedes. Daimo Samo villagers had significantly higher IgG and IgM values than healthy Mogadishu residents. Very high concentrations of IgG and IgM were demonstrated in sera from patients with visceral leishmaniasis. Somali patients with malaria also had marked hypergammaglobulinaemia, however, only in the IgG class. The high levels of IgG, IgM and IgA demonstrated in sera from Somalis, indicate the need for establishing local reference values and should be considered when introducing serological tests in tropical countries. Such methods are usually adopted to conditions in industrialized countries, where immunoglobulin contents of sera are lower.

Amebiasis in Nicaragua: class specific serum antibody responses.

With the aid of the indirect hemagglutination (IHA) test and IgG ELISA the antibody profile against E. histolytica in León, Nicaragua was investigated in 562 sera from individuals belonging to various age groups. The highest reactivity was invariably recorded in the age group 6-15 years where 48% were seropositive. Several sera reactive by either one of IHA and IgG ELISA were negative by the other test. The main reason for this seems to be reactivity in different Ig classes. Treatment with 2-mercaptoethanol reduced the titre level in 63 of the 66 sera tested. Immunofluorescence using an anti-IgM conjugate showed that 26 of 43 sera contained specific IgM-antibodies, indicating that also unspecific reactions are involved in the IHA test. A comparison was made between class-specific reactivity in three population groups: healthy residents, healthy cyst carriers and patients with recent or acute liver abscess. No significant difference in the prevalence of reactions above the diagnostic significance level was recorded between cyst carriers and healthy residents. However, among the cyst carriers 33% had IgA and/or IgM antibodies but no demonstrable specific IgG. Most patients with recent and all with acute liver abscess reacted significantly above the diagnostic limit in all three tests.

Antibody responses in schistosomiasis haematobium in Somalia. Relation to age and infection intensity.

Antibody responses in schistosomiasis haematobium were studied in relation to age and infection intensity in Somalia. The area is highly endemic for Schistosoma haematobium but free of S. mansoni. Antibodies of the IgG class against particulate antigens of S. mansoni adult worms were investigated by immunofluorescence (gut and somatic associated antigens) and against soluble egg and adult worm antigens by ELISA. Total IgE levels were examined by Pharmacia IgE RIA, and specific IgE against soluble adult worm antigen by enzyme immunoassay. The IgG antibody response showed a characteristic pattern with highest reactivity against both gut associated and soluble egg antigens in the age group 10-14 years, when both prevalence and intensity of the infection were highest. Reactivity against somatic associated antigen was also high in this age group, but it increased slightly and remained at high level in the older ages. It is thought that such antigen is exposed mainly after the death of the parasite and that the antigenic stimulation may remain throughout most of the life of infected individuals. On the other hand, the IgG antibody reactivity against soluble adult worm antigen was low during childhood, but it increased significantly with age. It is suggested that repeated booster effects are needed for more potent response against these antigenic components. The finding of high levels of total IgE already in the youngest age groups, together with low specific IgE response, indicates that mainly other antigens are involved in the IgE production. The specific IgE response against soluble adult worm antigen was low but increased significantly with age.

Antibodies against Entamoeba histolytica antigens in sera from individuals with amoebiasis of different localization.

The over-all contents and relative component composition of Entamoeba histolytica antigens in abscess fluids and in extracts of cultured amoebae, strain NIH 200, were studied by antigen-catching EIA, counterimmunoelectrophoresis (CIE) and immunoblotting techniques. The antigen contents of liver abscess fluid were determined semiquantitatively by the antigen-catching EIA in four cases. In CIE against a standard "diagnostic" extract of cultured amoebae, sera from cases of acute amoebic liver abscess gave 4-5 precipitation lines while sera from cases of intestinal amoebiasis gave at most 3 lines. In immunoblotting tests with the same antigen, intestinal cases gave blotting bands in the intermediate molecular weight range (25-99 kD) while acute abscess cases, in addition, gave bands in the high (100-175 kD) and low (= less than 25 kD) molecular weight range. These serological differences between clinical forms of amoebiasis were more definite when using amoeba abscess fluid as antigen. Amoeba antigens in high concentrations could be demonstrated in amoeba abscess fluids with all methods employed. In immunoblotting experiments abscess fluids generally gave stronger and more numerous bands with anti-amoeba antibody-containing sera than did the standard "diagnostic" antigen from cultured amoebae. Especially the abscess fluids gave with sera from acute abscess cases a number of prominent bands in the low molecular weight range (less than 25 kD). The experiments in this study were performed with crude amoebic extracts, which contained a multitudes of antigenic components and a still greater diversity of antigenically inert proteins.(ABSTRACT TRUNCATED AT 250 WORDS)

Incidence of toxoplasmosis in pregnant women in the city of Malmö, Sweden.

The incidences of latent and primary Toxoplasma gondii infections in pregnant women were studied using stored sera from 4,351 women delivered in the city of Malmö, Sweden in 1982 and 1983. Infants born to mothers with signs of primary infection (seroconversion or specific IgM in sera collected during pregnancy) were studied with regard to serological evidence of congenital infection (specific IgM in cord serum). Sera were tested for specific IgG antibodies by direct agglutination test and enzyme-linked immunosorbent assay, and for specific IgM by immunosorbent assay. 40% of the women were seropositive at delivery. Seroconversion (change from negative to positive serological status) was demonstrated in 12 pregnant women and specific IgM in the first postconceptional serum sample (indicating infection in the first trimester or in the last year(s) before conception) in another 17. The incidence of primary maternal infection was calculated to 4-6:1,000 deliveries. Among the 29 infants born to mothers with seroconversion or, alternatively, IgM in the first postconceptional sample, 6 had laboratory signs of congenital infection. One of the 6 had a positive toxoplasma isolation test in autopsy material and 5 had a clearly positive IgM value in cord serum. The real incidence of congenital infection in this material is unknown since IgM might be absent or sparse in cord serum in spite of congenital infection. The noticeable prevalence of maternal toxoplasmosis in Malmö calls for further incidence studies in Sweden.

Detection of giardia antigen in stool samples by a semi-quantitative enzyme immunoassay (EIA) test.

A semi-quantitative enzyme immunoassay (EIA) test for the detection of Giardia intestinalis antigen in faeces was developed. In order to avoid unspecific reactions due to anticalf serum activity, IgG fractions of anti-giardia rabbit and sheep sera were purified from antibovine antibodies by immunoadsorption. Faecal specimens tested in the assay were mixed with normal horse serum to avoid unspecific and proteolytic effects of stool components. Out of a range of bacterial and parasitic antigens tested, only high concentrations of Ascaris suum egg antigen and Shigella sonnei gave slight unspecific reactions. During an outbreak of waterborne giardia infection faecal samples from 49 individuals were tested. Giardia cysts were found by microscopy in 22 individuals and specimens from 24 persons were positive by EIA. The estimated amount of antigen found in the positive samples ranged from 50 to 5,000 ng giardia protein/ml. After filtration through a 0.45 microns filter antigenic activity was found in the filtrate of 10 samples. In 3/6 samples with no cysts detected by microscopy, mainly soluble giardia antigen was found.

The prevalence of Entamoeba histolytica in lactating women and in their infants in Bangladesh.

Entamoeba histolytica was studied in 33 lactating women and their infants in a periurban village in Bangladesh. Infant-mother pairs were followed for a period of 10-15 months: 67% of mothers excreted E. histolytica during the observation period, the majority for 3 months or more. Only one mother was symptomatic, with a mild, non-dysenteric diarrhoea. 58% of mothers were seropositive, several of them continuously and with a high titre, indicating past invasive infection. 67% had detectable antibodies in breast milk and 36% in saliva. Despite the high prevalence of E. histolytica in these mothers, infants were mostly uninfected: E. histolytica cysts in small numbers were found in only 2 of 1200 samples from infants aged 6 and 10 months. Several of the children were infected with Giardia lamblia. Although lower exposure to E. histolytica than to Giardia may account for the difference in infant infection rates with these parasites, defence mechanisms possibly exist which protect against E. histolytica but are ineffective against Giardia.

Human toxoplasmosis in Somalia. Prevalence of Toxoplasma antibodies in a village in the lower Scebelli region and in Mogadishu.

The acquisition of Toxoplasma antibodies in various age groups was studied in 2 populations in Somalia, inhabitants of a village in the southern part of the country and residents in Mogadishu. The overall prevalence of antibodies was 56% in the village and 40% in Mogadishu. In both populations, antibodies were acquired early in life. At the age of 10 years, 44% of the villagers and 31% of the Mogadishu children were seropositive. In Europe and the USA the highest rate of antibody acquisition has been recorded in young adults. This difference may indicate different routes of transmission. In Europe and the USA Toxoplasma is transmitted mainly by the ingestion of undercooked pork or lamb. Undercooked meat is seldom consumed in Somalia, and pork not at all. However, the soil in Somalia is heavily contaminated with cat faeces and the humid climate in the southern part of the country may contribute to long survival of oocysts. In the villages all household activities are performed on the ground and in Mogadishu children play mainly outdoors on the ground. It therefore seems that conditions in Somalia favour transmission by oocysts rather than by infected meat. The early acquisition of antibodies in Somalia suggests that infection during pregnancy and, therefore, congenital toxoplasmosis are rare.

Amoebicidal activity of the antifungal antibiotic sinefungin against Entamoeba histolytica.

The antifungal antibiotic sinefungin, a potent inhibitor of methyl transferases and a potential inhibitor of polyamine biosynthesis, exhibited marked cytolytic activity against Entamoeba histolytica in vitro with respect to all the amoeba strains tested. Strains of high and low virulence displayed equal sensitivity to the antibiotic. Minimal Amoebicidal Concentration was 5 to 10 micrograms/ml.