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Letrazuril may have a short term effect in treating cryptosporidiosis in AIDS patients. Cryptosporidiosis is a severe diarrhea caused by a parasite which often causes or contributes to mortality in AIDS patients. An oral dose of 50 milligrams/day of letrazuril was given to 35 AIDS patients with cryptosporidiosis. Before treatment, 74% of the patients had five or more bowel movements a day, and 91% had watery bowel movements. Of the 35 patients, 12% did not respond to treatment, 63% experienced at least a 50% decrease in bowel movements per day, and 3% experienced a reduction to two or fewer bowel movements per day. The drug eradicated the parasite in 10 (40%) of 25 of the patients whose stools were tested. An average of 1.7 weeks of treatment caused a response among those whose condition improved. However, in 65% of the responders the disease relapsed an average of 1.2 months after treatment began. The major side effect was a rash which occurred in 20% of the patients and caused them to cease treatment.

Cytomegalovirus (CMV) infection may cause retinitis or inflammation of the retina, the inner lining of the eye; this is the most common cause of blindness in patients with acquired immunodeficiency syndrome (AIDS). The drug foscarnet was shown to inhibit viral deoxyribonucleic acid (DNA) polymerase, an enzyme involved in the formation of the viral DNA molecule which carries the genetic information of the CMV microorganism. Studies suggest that foscarnet may be effective in preventing the progression of CMV retinitis. The effectiveness of foscarnet was assessed in 17 patients with CMV retinitis. Foscarnet produced a complete response in eight patients, a partial response in eight patients, and no response in one case. The excretion of CMV in the urine was not decreased by foscarnet in 4 of 12 patients, despite improved retinal lesions. Adverse effects of foscarnet included: kidney failure, reflected by increased blood creatinine levels in 50 percent of patients and also the need for dialysis, the artificial filtration of the blood, in two patients; and anemia resulting in the need for blood transfusion in 10 patients. Among seven patients maintained on foscarnet therapy, four suffered recurrence of CMV retinitis, and only one remained in remission for longer than 24 weeks. The drug zidovudine did not worsen the incidence of side effects due to foscarnet use. Thus, foscarnet is effective in suppressing the activity of CMV in retinitis, although the suppression is not maintained. (Consumer Summary produced by Reliance Medical Information, Inc.)

Chronic fatigue syndrome CFS is a severely disabling illness of uncertain aetiology. It is characterized by a chronic, sustained or fluctuating sense of debilitating fatigue without any other known underlying medical conditions. It is also associated with both somatic and neuropsychological symptoms. Both physical and laboratory findings are usually unremarkable. Regional cerebral blood flow rCBF was assessed in 60 clinically defined CFS patients and 14 normal control NC subjects using 99Tcmhexamethylpropyleneamine oxime 99TcmHMPAO single photon emission computed tomography SPECT. Compared with the NC group, the CFS group showed significantly lower corticalcerebellar rCBF ratios, throughout multiple brain regions P< 0.05. Fortyeight CFS subjects 80 showed at least one or more rCBF ratios significantly less than normal values. The major cerebral regions involved were frontal 38 cases, 63, temporal 21 cases, 35, parietal 32 cases, 53 and occipital lobes 23 cases, 38. The rCBF ratios of basal ganglia 24 cases, 40 were also reduced. 99TcmHMPAO brain SPECT provided objective evidence for functional impairment of the brain in the majority of the CFS subjects. The findings may not be diagnostic of CFS but 99TcmHMPAO SPECT may play an important role in clarifying the pathoaetiology of CFS. Further studies are warranted.

We have purified pili from isogenic opacity colony variants that were derived from 14 gonococcal strains. Pili purified from opaque colonies of one strain usually differed from pili purified from transparent colonies of the same strain. In 10 of the 14 strains examined, the apparent subunit molecular weight of pilin isolated from the opaque variants was larger than that seen with pilin obtained from transparent variants. In addition there were demonstrable intra-strain differences in the isoelectric point and buoyant density of pili derived from the opacity variants. Because gonococci express differing opacity phenotypes during the menstrual cycle, it is possible that the pili of these organisms may also alter in vivo.

P pilus filaments are important in binding to globoside through an adhesin located at the tip of the pilus. There is considerable antigenic variation among P pili, and the immunologic response is usually serotype specific. We purified denatured pilin subunits and used them as immunogens to prepare more broadly cross-reactive antisera. Although antifilament antisera (AFA) detected predominantly the homologous strain, antisubunit antisera (ASA) prepared from two different strains detected P pili in 16 of 16 and 14 of 16 P-piliated strains by Western blotting (immunoblotting). The binding of ASA to the homologous pilus filament was inhibited by only 3 of 17 strains. ASA agglutinated only two of nine heterologous strains and immunoprecipitated pili from one of three heterologous strains. By immunoelectron microscopy ASA was seen to bind to pilus filaments but not as strongly as AFA. Antiserum raised to the denatured pilin subunit was not substantially more reactive with pilus filaments derived from heterologous strains than was AFA. ASA was, however, a very useful probe for detecting most P pilins.

Carrier strains of Neisseria meningitidis are recovered almost solely from the posterior pharynx and they are often nongroupable or rough. Invasive strains can be serogrouped (encapsulated). We studied adherence of both carrier- and patient-derived serogroupable and nongroupable meningococci to buccal epithelial and posterior pharyngeal cells. Fresh meningococcal isolates attached significantly better to pharyngeal than to buccal cells (P = 0.01). Strains that could be serogrouped adhered less than nongroupable strains (P less than 0.05). Meningococci passed in vitro became hemagglutinin negative and nonpiliated. Hemagglutinin-negative meningococci always adhered less to both epithelial cell types than the hemagglutinin-positive variants of the same strain. These results indicate that meningococcal pili probably mediate attachment to oropharyngeal cells, but encapsulation may reduce adherence. Localization of meningococci in the posterior pharynx is in part explained by the receptivity of the epithelial cells in this area for meningococci.

Bacteria which attach to different mucous membranes should have differing specificities of adherence in vitro. Human Escherichia coli isolates from blood and urine (pathogens) and from stool and throat (commensals) were characterized as to the patterns of hemagglutination (HA), as well as the structure and function of their pili. Bacterial HA was done in microtiter plates and on slides after bacterial growth in broth or agar. Human erythrocytes were agglutinated by 95% of the pathogens and 65 to 70% of the commensals grown in broth or agar. Mannose-resistant HA was characteristically caused by pathogens, and commensals characteristically caused mannose-sensitive HA of guinea pig cells. Strains often had both mannose-resistant and mannose-sensitive reactions, or even a mannose-paradoxical reaction. Pathogens more often caused HA, but titers were lower than those for commensals. Slide HA was less sensitive than the microtiter method. All isolates were piliated. Commensals also had more pili than pathogens when grown in broth (117.8 versus 38.3 pili per bacterium), but pathogens had more pili after growth on agar (32.1 versus 8.1 pili per bacterium). Isolates causing high-titer HA had large numbers of pili (greater than 85 pili per bacterium), but some well-piliated strains were non-hemagglutinating. Pili were purified from seven E. coli strains from different sites of isolation and with different erythrocyte-binding specificity. Pili usually migrated as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, more than one type of pilus could be copurified from some strains since there were two or more bands after separation in octyl-glucoside and two different amino terminal sequences. Protein sequencing was done on five different pili: four resembled type 1 pili and one was a P fimbria. The type 1-like pili (strains 2239 and 9353) had an initial variable sequence of 1 to 5 residues, followed by a common region of 21 residues. The P fimbria (strain 7714) had different erythrocyte-binding specificity but was still 27% homologous with 2239 and 9353. E. coli strains from different body sites have characteristic attachments to erythrocytes. Pili derived from these different sources may also have different binding specificity, but they are similar in primary structure.

P pili (or fimbriae) are present on most pyelonephritogenic Escherichia coli strains, and they mediate binding to erythrocytes and epithelial cells. To determine the antigenic diversity of P pili, we purified the pili from 14 bacteremic E. coli strains which caused mannose-resistant hemagglutination. Pilus preparations consisted of one to three bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis and ranged in molecular weight from 14,000 to 19,500. There was no single band common to all the strains. An enzyme-linked immunosorbent inhibition assay detected 20 ng of pilus antigen. When four different rabbit antisera were used, only two or fewer heterologous strains could inhibit the enzyme-linked immunosorbent assay. Immunoblots yielded the same results. Protein sequences of four P pili had identical N termini. These results show that despite having identical amino-terminal sequences, P pili are antigenically heterogeneous. The receptor-binding domains which are likely to be identical in all strains must be immunorecessive.

Many enterobacteria can cause agglutination of erythrocytes, but previous investigations have not proven which components of the bacteria are responsible. We used a strain of Escherichia coli K12 which causes mannose-sensitive hemagglutination (HA) of guinea pig cells. Common pili were purified from these bacteria by shearing them from the bacteria followed by selective precipitation in acid and ammonium sulfate. Isopycnic centrifugation in cesium chloride removed the remaining outer membrane protein contaminants. These pili are pure by electron microscopy and gel electrophoresis. By amino acid analysis, they have a mol wt of 17,099 and consist of 45% nonpolar residues. These purified pili agglutinate guinea pig erythrocytes, a reaction that is inhibited by anti-pili antibodies and by saccharides related in structure to D-mannose. Proteolytic treatment of erythrocytes does not diminish HA but rather increases the pili-induced HA of human cells. Neuraminidase enhances HA and mannosidase slightly diminishes it. It is concluded that purified pili alone cause HA of erythrocytes by binding to mannose-like molecules on the erythrocyte surface. Thus HA by bacterial pili serves as a useful model system for the mechanism of bacterial pili attachment ot cell membranes.

We have demonstrated binding of purified pili from a strain of Escherichia coli to Vero cell monolayers as a model of prokaryotic-eukaryotic cell adherence. Pili bound to the tissue culture in a rapid reaction that did not require enzymatic activation. Attachment occurred optimally at pH 4-5 and could be inhibited by analogues of D-mannose, anti-pili antibodies, or by preincubation of tissue cells with mannose-specific plant lectins. Binding remained after treatment of the monolayer with glycosidases, trypsin, or a protease mixture but was enhanced after neuraminidase treatment. These results indicate that bacterial binding can occur via pili which act like lectins and presumably bind to mannose-containing glycoproteins on mammalian cell surfaces.

A more complete understanding of meningococcal disease has been hampered by the lack of an appropriate animal model. Previous models have utilized injections of meningococci, which precludes the study of nasopharyngeal colonization and invasion. We have developed a model for meningococcal disease in which litters of 5-day-old mice are challenged intranasally with 10(7) viable meningococci. Bacteremia is monitored by jugular venous blood cultures, and cerebrospinal fluid is sampled by cisternal punctures. Human disease-associated and carrier strains were compared; nasopharyngeal colonization was similar for these bacteria, but the case-associated strains were much more frequently invasive and caused bacteremia. Twenty-one percent of bacteremic animals had meningitis. There was an age-related susceptibility to infection which correlated inversely with the levels of serum complement. Preinjection of iron dextran increased the number of animals which were bacteremic, the concentration of bacteria in blood, and nasopharyngeal colonization. Noncapsular variants of virulent meningococci did not colonize nasopharyngeal tissue in vivo, and they were not invasive. This neonatal mouse model mimics meningococcal disease as seen in humans and may be useful in studying the initial events in the pathogenesis of meningococcal disease.

We have shown previously that some strains of Pseudomonas aeruginosa from patients with cystic fibrosis are phagocytized by human polymorphonuclear leukocytes in the absence of serum opsonins. The purpose of this study was to identify the bacterial features which render certain strains susceptible to nonopsonic phagocytosis. Three strains were phagocytized by human neutrophils and monocyte-derived macrophages, and two were not, as determined by luminol-enhanced chemiluminescence, visual inspection of stained smears, and bactericidal assay. Strains that were phagocytized formed pellicles when grown in static broth, but the phagocytosis-resistant strains did not. The phagocytosis-susceptible strains were more heavily piliated and more hydrophobic than the resistant strains. Bacteria exposed to heat (60 degrees C) or UV irradiation were depiliated, as assessed by electron microscopy, and rendered resistant to phagocytosis. When P. aeruginosa was grown on agar, it was piliated, hydrophobic, and susceptible to nonopsonic phagocytosis, but when grown to stationary phase in shaken broth, it was nonpiliated, less hydrophobic, and resistant to phagocytosis. It appears that nonopsonic phagocytosis of certain P. aeruginosa strains by human polymorphonuclear leukocytes and macrophages is facilitated by hydrophobic interactions which may be determined in part by pili.

Previous studies have noted that the prevalence of certain gonococcal colony types is influenced by the physiological state of the human host. Consequently eight different gonococcal strains were passed on clear typing medium, and opaque (Op) and transparent (Tr) variants of heavily pilated (P++) bacteria were selected. P++Op and P++Tr were injected into 11-day-old chicken embryos, and the resulting mortality was determined. In every case the Tr variants were more lethal to the embryos than were the Op variants (P = 0.01). P++Tr were stable in vivo, but chicken embryos injected with P++Op strains developed bacteremia with P++Tr organisms. However, chicken blood was not bactericidal for either colonial variant. These results indicate that transparent pilated bacteria may be more virulent than opaque pilated bacteria and that in vivo selection of transparent forms occurs.