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Hemolysis-in-gel (HIG) test for antibodies to influenza A, measles and mumps using liquid nitrogen freezed erythrocytes coupled with the respective viral antigen.

A drawback with the hemolysis-in-gel test is the constant need for fresh erythrocytes which must be treated with virus before incorporation in the gel. This problem was overcome by freezing small droplets containing erythrocytes to which antigen had been attached. The droplets were stored at -70 degrees C or in liquid nitrogen. This modification was applied in detecting antibodies to influenza, measles and mumps viruses and the results were shown to equal those obtained with fresh erythrocytes.

Animals

Inversion homozygosity of chromosome no. 9 in a higly inbred kindred.

A pericentric inversion of chromosome no. 9 was present in seven of 10 members of a highly inbred kindred investigated; two were inversion homozygotes and five were heterozygotes. Inversion homozygosity was observed in both the propositus, ascertained because of ambiguous genitalia, and his phenotypically normal father. A phenotypically normal sister and brother with similar clinical findings proved to be inversion heterozygotes. These findings conclude that no causal relationship exists between the inversion and the abnormal phenotype.

Adult

Hemolysis-in-gel test in immunity surveys and diagnosis of rubella.

The hemolysis-in-gel (HIG) technique was adapted for rubella antibody determinations. Use of sucrose gradient purified virus and its coupling with CrCl3 to chicken erythrocytes resulted in gel plates that could be stored for several weeks and were suitable for reproducible antibody determinations. In a serological survey of young healthy adults the HIG values (range less than 2--13 mm) were in close correlation to those obtained by the HI test (less than 10 5o 320). The Hig test seems well suited for screening the need of vaccination. Seronegative sera (HIG less than 2,HI less than 10) gave without heat inactivation hemolysis zones ranging from 4 to 6.5 mm. Although the present rubella HIG test did not measure IgM antibodies, the test, by virtue of its accuracy and sensitivity--extending to antibody levels corresponding to HI titers 2--10--provides a simpler and more rapid means for diagnosis of rubella infections than the conventional HI and CF tests.

Adult

Antibodies to human coronavirus OC43 measured by radial haemolysis in gel.

An application of the haemolysis-in-gel (HIG) technique was developed to quantitate antibodies against the human coronavirus OC43. Preinfection and convalescent sera from two patients with verified OC43 infection showed a significant increase in antibody titres measured by HIG as well as by haemagglutination-inhibition (HI). 241 of the other 306 sera tested (80%) caused radial haemolysis in gels containing viral antigen-sensitized erythrocytes and complement. No haemolysis was seen in gels prepared with unifected material. Correlation of the diameter of the haemolysis ring to the respective HI titre was highly significant. However, 62 sera (20%) with HI titres between 8 and 320 were negative in the HIG. Attempts to show that these sera contained nonspecific inhibitors of haemagglutination were unsuccessful. OC43 HI and HIG probably measure different antibody ppulations.

Antibodies, Viral

OC43 strain-related coronavirus antibodies in different age groups.

Serum antibodies against human coronavirus OC43 in different age groups were measured by complement fixation (CF), haemagglutination inhibition (HI), radial diffusion haemolysis-in-gel (HIG), and solid-phase radioimmunoassay (RIA) methods. Antigen grown in suckling mouse brain was used in all tests. Results obtained by the CF and HIG tests, and the RIA, were in good agreement with regard to the presence or absence of antibodies. Similar results were also obtained with the HI test if nonspecific haemagglutination inhibitors were first removed by treatment with phospholipase C and only titers of 1:20 or greater were considered positive. Children 6--23 months of age (n = 45) were without measurable coronavirus antibodies in all four assays. A rapid increase in the prevalence of antibodies then occurred in subsequent age groups, and practically all persons 6 years of age or older were found to have OC43 antibodies as measured by the HIG test or the RIA. The mean antibody levels determined by these two methods continued to increase, however, up to the age group of 10--14 years. This increase in antibody levels after the initial antibody incidence plateau may be due to boosting effects caused by related coronavirus strains, since OC43 antigens are known to cross-react with antibodies induced by other human coronaviruses. Taken together, these data suggest that OC43 virus, or an antigenically related coronavirus strain, is very common in Finland.

Adolescent

[Hemolysis in ger test -- an alternative to hamagglutination inhibition test for search of antibodies against rubella virus (author's transl)].

640 sera of unknown immune status were tested in the rubella hemagglutination inhibition test (HI) as well as in hemolysis in gel test (HIG). The results have been analysed by statistical methods. The reproducibility of zones of hemolysis are 1.7 mm for the long term precision and 1.0 mm for the short term precision. Comparing the HIG and the HI tests we had a correlation of 97% = sensitivity (544 out of 561 sera) in positive reacting sera and of 87% = specifity (69 out of 79 sera) in the negative reactions. We consider a person to have antibodies against rubella if the zone of hemolysis had a diameter of 8 mm or above. In cases of diameters under 8 mm the results of IH and HIG-tests do not correlate. The reasons are not completely known and should be investigated.

Antibodies, Viral

Nonspecific inhibitors of coronavirus OC43 haemagglutination in human sera.

Antibodies against the human coronavirus OC43 in human sera were measured by haemagglutination inhibition (HI), complement fixation (CF), and radial diffusion-haemolysis in gel (HIG) techniques. The apparent HI titres in a fraction of sera with no antibodies detectable by the two other methods were found to be reduced considerably after treating the sera with phospholipase C (PLC). The PLC treatment also reduced the apparent HI titres in some sera containing variable amounts of CF and/or HIG antibodies, but did not affect antibody determinations by the two latter methods. These results suggest that false positive results can be obtained in the OC43 HI test unless the sera are treated with phospholipase C before the assay.

Antiviral Agents

Synergistic effects of antiviral agents and humoral antibodies in experimental Herpesvirus hominis encephalitis.

Experimental Herpesvirus hominis encephalitis in 3-week-old Swiss mice was utilized to evaluate the antiviral effects of adenine arabinoside (ara-A), phosphonacetic acid (PAA), and humoral antibodies to H. hominis (HIG). Administration of ara-A, PAA, or HIG during the early phase of infection reduced the mortality rate of H. hominis encephalitis in mice. Combined administration of humoral antibodies with an antiviral agent (ara-A or PAA) resulted in an enhanced protection, accompanied by a decreased virus concentration in the brain tissues and a diminished tissue injury. The generality of this enhancing effect of humoral antibodies on the action of antiviral agents and its possible use in therapy of viral infections warrant further investigation.

Animals

[Use of hemolysis in gel in the serodiagnosis of rubella. Comparison with hemagglutination inhibition].

The hemolysis in gel test (HIG) was compared with the hemagglutination inhibition test (HI). This new test is simple and time-sparing since it does not require dilution or serum pre-treatment, and can be measured directly in mm. It is not more expensive than the HI method. It has proved sensitive and is not affected by non specific serum hemagglutination inhibitors and is conclusive whenever HI titers are looked upon with suspicion (titers of less than 1/20). It is therefore well adapted to mass screening of immunity against rubella but in the field of recent infections HI method still has its role to play. Using the HIG test as a reference, we measured the loss of Ig to rubella following the three most commonly employed methods for removal of inhibitors: the mean values of 16 sera were 25% of antibodies lost after kaolin pre-treatment, 62% lost after heparine-Mn-Cl2 procedure 75% lost after dextran-CaCl2 treatment.

Hemagglutination Inhibition Tests

Mixed Müllerian tumors of the uterus. Ultrastructural studies on the differentiation of rhabdomyoblasts.

In the present study the differentiation of rhabdomyoblasts of two carcinosarcomas of the uterus is analysed electronmicroscopically. During the development of rhabdomyoblasts three cell types can be distinguished: 1. the undifferentiated mesenchymal cell with abundant ribosomes but few other cell organelles. Usually these cells are already associated to each other in strands of 2-4 cells. Occasionally cytoplasmic areas with numerous nonspecific filaments can be observed. 2. the light rhabdomyoblast which is characterized by its conspicuous nonspecific cytoplasmic filaments and its reduction in ribosomes. Specific myofilaments can be visualized only occasionally in this cell type. 3. more differentiated rhabdomyoblasts. These cells can be identified clearly by the presence of large numbers of myofibrils and myofibrillar fragments. Higly organized myofibrils as seen in skeletal muscle are seldom found. Transitional forms between these cell types are also observed. The functions of Z-bodies and of the sarcotubular system in the process of myofibrillar differentiation are discussed.

Aged

Autoradiographic localization of opiate receptors in rat brain. I. Spinal cord and lower medulla.

The localization of opiate receptors in the spinal cord and lower medulla has been elucidated by the autoradiographic identification of stereospecific [3H]diprenorphine (a potent opiate antagonist) binding sites. The opiate receptors were higly localized to: layers I (marginal cell zones) and II (substantia gelatinosa) of the dorsal horn of the spinal cord; the substantia gelationsa of the spinal trigeminal nucleus; components of the vagal system, including the vagus nerve, nucleus tractus solitarius, nucleus commissuralis, nucleus intercalatus, nucleus ambiguus and nucleus originis dorsalis vagus; the area postrema. Examination of [3H]etorphine (a potent opiate agonist) binding sites showed the same distribution. We conclude that, in these brain regions, opiate receptors are (1) highly associated with areas receiving small, afferent primary fibers, (2) strategically placed to modulate noxious stimuli as well as explain some visceral side effects of opiate administration.

Afferent Pathways

A simple and sensitive nonradioactive method for the detection of urinary human chorionic gonadotropin and diagnosis of early human pregnancy. II. Single-unit test.

A simple, sensitive, and reliable single-unit nonradioactive method for the detection of human chorionic gonadotropin (hCG) in concentrated urine and the diagnosis of early pregnancy is reported. This unit, presently termed the Ayerst pregnancy test kit (APTK), consists of four components: a sampler-filter paper cone, an ultrafilter-concentrator to which a vial holder is attached, a support stand with a mirror, and an immunologic reagent vial. In the APTK, 5 to 6 ml of urine were sampled, filtered, and concentrated, and the hCG in the retentate was detected by Ayerst immunologic reagents [APTK(AY)] and by the Pregnosticon "All In" [APTK(P)]. Some of the unconcentrated urine samples (0.1 ml) were also tested in hemagglutination inhibition tests (HIT) using Ayerst [HIG(AY)] and Pregnosticon "All In" [HIT(P)] reagents. Urine samples from pregnant, nonpregnant (ovulating and nonovulating), perimenopausal, and menopausal women were tested. It was found that the APTK(AY) and APTK(P) were significantly more sensitive and reliable than the HIT(AY) and HIT(P) in detecting low levels of urinary hCG for early diagnosis of pregnancy. The sensitivity and specificity of the APTK(AY) were better than those of the APTK(P). The APTK(AY) give significantly more correct positive and negative results than the other tests performed simultaneously. The APTK(AY) is simpler and safer than the serum radioimmunoassays and radioreceptor assay presently used to detect low levels of hCG for the early diagnosis of pregnancy and other hCG-producing states.

Chorionic Gonadotropin

[Detection and titration of antitetanus antibodies using an automated passive hemagglutination method].

An automated reversed passive haemagglutination technique is described, using human erythrocytes sensitized by the chromic chloride method with higly purified tetanus toxoid. Such erythrocytes are agglutinated by the specific antibodies in a Technicon autoanalyzer. Clots are decanted, supernatant is hemolyzed and optical density is measured at 550 millimicron. Assay standards for comparison, ranging from 5 to 25 UI/ml, are prepared from human antitetanus immunoglobulins titrated by toxin neutralisation assay in mice. This method allows to screen donors' sera for presence of high titers of tetanus antibodies (larger than or equal to 5 UI) suitable for preparation of antitetanus immunoglobulins. 4,4% of donors have circulating antitetanus antibody levels corresponding to 5 UI/ml or more, by this method. The specificity of antibodies has been confirmed by the neutralisation assay in mice. The results obtained among 1.000 donors well agree with those of the counter-immuno-electrophoresis technique (C.E.P.). But, when, compared with C.E.P., the haemagglutination assay appears more objective, more quantitative and sensitive, and allows to get not only a rapid screening test but also a precise titration simultaneously.

Antibodies, Bacterial

Thiamin deprivation in ventromedial hypothalamic hyperphagic rats: anorexia, specificity of food aversion, and a dietary consideration.

The anorexic consequence of thiamin deprivation was investigated in ventromedial hypothalamic (VMH) hyperphagic rats under either high-fat or low-fat thiamin-free diet conditions. The low-fat diet maintained feeding significantly longer in thiamin-deprived VMH rats than in intact rats, whereas the hig-fat diet sustained feeding in thiamin-deficient intact rats and accelerated anorexia onset in vitamin B1 deprived VMH rats. This effect was noted under both ad lib and pair-feeding conditions. Thiamin-deprived VMH rats subjected to weight control developed anorexia sooner than intact subjects regardless of the diet employed. The VMH rats fed a high-fat diet failed to resume feeding after thiamin readministration, which was interpreted as a permanent aversion to this diet. The relation between dietary intake and conditioned taste aversion is discussed with reference to the VMH and intact rat.

Animals

N6-methyladenine DNA modification modulates pathogen virulence in nematodes.

Understanding the global regulatory mechanisms that control pathogen virulence gene expression is essential for elucidating the molecular basis of pathogenicity. N6-methyladenine (6 mA) plays a crucial role in regulating gene expression in response to various environmental stresses; however, its role in pathogen virulence remains largely unexplored. Here, we report the widespread occurrence of 6 mA across 17 nematode isolates and map its genomic landscape in six notorious agriculturally important pathogen root-knot nematodes (RKNs). We demonstrated that 6 mA is characterized by a conserved GAG motif across nematodes, but exhibits species-specific distribution patterns and distinct effects on gene expression. In particular, its enrichment in transposable elements (TEs) differs between polyploid and diploid nematodes, suggesting lineage-specific epigenetic regulation potentially associated with polyploidy. We further identified two functional 6 mA demethylases, MiNMAD-1 and MiNMAD-2, and confirmed their catalytic activity and active sites. Host-induced gene silencing (HIGS) of minmad-1 significantly increased plant resistance to three polyploid RKN species. A detailed functional analysis revealed that minmad-1 knockdown broadly affected gene expression during the parasitic stage, including genes involved in virulence, thereby reducing nematode infectivity. Together, our findings suggest 6 mA demethylase as a key epigenetic regulator of RKNs' virulence, providing new insights into nematode biology and offering promising targets for the development of sustainable control strategies.

Animals

Post-ischaemic paraesthesia in pellagrins.

A quantitative assessment of post-ischaemic paraesthesiae has been made in 50 pellagrins and 20 healthy identical controls. The results show a higly significant diminution of the paraesthetic response in pellagrins. In pellagrins having peripheral neuropathy the depression of paraesthesiae was more marked than in those without peripheral neuropathy. There was no consistent relationship between severity of peripheral neuropathy and degree of depression of paraesthetic response.

Adolescent

Homeostatic regulation of epidermal cells in culture.

The growth potential (DNA synthesis) of purified epidermal basal cells in culture, according to time reveals two peaks, at days 1 and 6. The same potential estimated in total cultures of epidermal cells (including basal and high level cells) shows only one peak at day 9. This suggests that hig-level mature cells inhibit DNA synthesis in basal cells in culture. This interpretation has been confirmed in showing that increasing amounts of mature cells inhibited DNA synthesis in basal cells in a dose dependent manner. Basal cells in culture are therefore susceptible to homeostatic regulation. However, this regulation is not tissue specific since DNA synthesis was also inhibited by mature epidermal cells in cultures of dermal as well as bone marrow cells. Assuming that the culture of epidermal cells is representative of the in vivo situation, these experiments would suggest that the specificity of action of the G1 chalone in vivo might depend as much on the target tissue as the proper nature of the inhibitor.

Animals