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Susceptibility of inbred rat strains to experimental thyroiditis: quantitation of thyroglobulin-binding cells and assessment of T-cell function in susceptible and non-susceptible strains.

Ten inbred strains of rats were immunized with crude homologous thyroglobulin emulsified in Freund's complete adjuvant in order to investigate strain susceptibility to the induction of both thyroiditis and antibody to thyroglobulin. Two strains (LH and AUG) were found to be extremely susceptible and had 100% incidence of thyroid lesions which in general varied from moderate to very severe (mean index of pathology+/-SE, 2-5+/-0-2 and 2-1+/-0-4 respectively). One other strain (HL) also had 100% incidence of lesions but there were consistently mild in character (1-1+/-0-1). Two strains (DA and SD) were variable, with thyroid change varying from negative to severe. Three strains (LEW, WAG and PVG/c) had occasional lesions and the remaining two strains (AS and CAM) showed no thyroid change. Four strains (LH, AUG, HL and DA) consistently produced good antibody responses to thyroglobulin (mean titres+/-SE 7-3+/-0-3, 9-5+/-0-4, 6-9+/-0-3 and 6-6+/-0-5 respectively). In contrast WAG and CAM rats failed to develop autoantibody and the responses of AS, PVG/c and SD strain rats were quite variable. Although the autoantibody response generally correlated well with the presence of thyroiditis in a particular strain, LEW, AS and PVG/c rats often had good antibody levels with minimal thyroid lesions. Females of the most susceptible strains (LH and AUG) were found to have significantly more severe thyroid lesions and higher antibody titres to thyroglobulin than males. The most susceptible strains were all found to be of the Ag-B5 major histocompatibility genotype whilst the least susceptible were of the Ag-B2 genotype. However, wide interstrain variability was noted within the Ag-B5 genotype particularly with respect to the induction and extent of thyroid lesions. It was not found possible to relate the divergence in susceptibility between rat strains of Ag-B5 and Ag-B2 genotypes to differences in respective numbers of thyroglobulin-binding cells within the circulation of the non-immunized animal. Similarly, there were no differences in response between a susceptible (LH) and non-susceptible (CAM) strain to the phytomitogens PHA and Con A.

Animals

Paramyxovirus-avian cell relationship: discrepant impact of 6-azauridine on virus production by susceptible and less susceptible cells.

The replication of mumps virus in susceptible chicken embryonic heart cells was escalated by daily treatment of cultures with 6-azauridine (6-AU). On the other hand, virus production by less susceptible liver cells was depressed by 6-AU. The population of infected susceptible cells was not increased, but the release of virus by infected susceptible cells was enhanced 20-fold by 10 mug of 6-AU per ml. Less susceptible cells, which incorporated less ribonucleic acid and protein precursor than susceptible cells, sustained a constant level of viral release during 6-AU treatment; however, the number of infected less susceptible cells underwent substantial decline.

Animals

Susceptibilities of penicillin-susceptible and -resistant strains of Streptococcus pneumoniae to RP 59500, vancomycin, erythromycin, PD 131628, sparfloxacin, temafloxacin, win 57273, ofloxacin, and ciprofloxacin.

The MICs of four new quinolones, sparfloxacin (AT-4140, CI-978), PD 131628 (the active form of the prodrug CI-990), temafloxacin, and Win 57273, compared with those of ciprofloxacin and ofloxacin were tested against 53 penicillin-susceptible, 35 penicillin intermediate-resistant, and 51 penicillin-resistant pneumococci. Susceptibility to RP 59500, a new streptogramin, was also tested and compared with those to the quinolones, erythromycin, and vancomycin. All MICs were determined by a standardized agar dilution method by using Mueller-Hinton agar supplemented with sheep blood. Quinolone, vancomycin, and RP 59500 susceptibilities were not affected by susceptibility or resistance to penicillin. For Win 57273, the MICs for 50% (MIC50) and 90% (MIC90) of strains tested were 0.015 and 0.03 micrograms/ml, respectively. MIC50S of both sparfloxacin and PD 131628 were 0.25 micrograms/ml, and MIC90S were 0.5 micrograms/ml. The MIC50 of temafloxacin was 0.5 micrograms/ml, and the MIC90 was 1.0 micrograms/ml. By comparison, ofloxacin and ciprofloxacin both yielded MIC50S of 1.0 micrograms/ml and MIC90s of 2.0 micrograms/ml. RP 59500 yielded an MIC50 of 0.5 microgram/ml and an MIC90 of 1.0 microgram/ml and was only 1 doubling dilution less active against 17 erythromycin-resistant strains. Vancomycin was active against all strains (MIC50, 0.25 microgram/ml; MIC90, 0.5 microgram/ml). All four experimental quinolones as well as RP 59500 show promise for therapy of infections with penicillin-resistant and -susceptible pneumococci.

Anti-Infective Agents

Susceptibility of anaerobic bacteria to sulfamethoxazole/trimethoprim and routine susceptibility testing.

The minimal inhibitory concentrations (MICs) of sulfamethoxazole and trimethoprim against 144 strains of obligately anaerobic bacteria were determined on Diagnostic Sensitivity Test agar (Oxoid) or in prereduced Diagnostic Sensitivity Test broth, both supplemented with sodium pyruvate (1 mg/ml), hemin (5 mug/ml), and vitamin K(1) (1 mug/ml). Fifty-eight percent of the strains were susceptible to sulfamethoxazole alone (MIC </= 16 mug/ml), only 12% were susceptible to trimethoprim alone (MIC </= 1 mug/ml), and 85% were susceptible to the combination of sulfamethoxazole plus trimethoprim (MIC </= 16 mug/ml) at a ratio of 19:1. All 45 strains of the Bacteroides fragilis group were susceptible to the combination. Synergy of the combination was often observed by a checkerboard MIC determination of 123 strains, usually most markedly when the ratio of the two components was near 1:1. However, there was also synergism at the ratio of sulfamethoxazole to trimethoprim of 16:1 in 61 (53.5%) of the 114 strains that could be evaluated for synergistic activity. When the strains were tested by the broth-disk test proposed by Wilkins and Thiel, modified by using prereduced Diagnostic Sensitivity Test broth instead of brain heart infusion broth and by using a smaller inoculum, there was over 90% correlation with the MICs. Poor results were found when the broth-disk tests were performed in brain heart infusion broth. There was very poor correlation between inhibition zone diameters by an agar diffusion method and MICs.

Bacteria

Discrepancies in isoniazid susceptibility profiles: Bactec MGIT 960-resistant but GenoType MTBDRplus-susceptible Mycobacterium tuberculosis strains in Hunan, China.

UNLABELLED: Discordant drug susceptibility testing (DST) results between the Bactec MGIT 960 system (MGIT) and the GenoType MTBDRplus assay (MTBDRplus) for isoniazid (INH) complicate clinical decision-making. In this study, we performed minimum inhibitory concentration (MIC) assays and whole-genome sequencing (WGS) on 53 Mycobacterium tuberculosis strains identified as INH-resistant by MGIT but INH-susceptible by MTBDRplus. The variants conferring INH resistance were evaluated by the WHO mutation catalogue. Our results showed that only five strains carried variants classified as "associated with resistance" (Group 1/2), including katG Trp39STOP, katG Ser315Asn, inhA -154G>A, and inhA Ser94Ala. In addition, 44 strains carried 70 variants classified as "Group 3: Uncertain significance" across nine genes, including katG, ahpC, inhA, Rv0010c, Rv1129c, Rv2752c, mshA, dnaA, and Rv1258c. The remaining four strains carried no variants (Groups 1-3) linked to INH resistance. No significant difference in the prevalence of high-level INH resistance was observed between lineage 2 and lineage 4 strains (&#x3c7;&#xb2; = 0.232, P = 0.630). Our findings indicate that the variants classified as "uncertain significance" may be the main genetic determinants causing discordant results, highlighting their associations with INH resistance that need to be further investigated. IMPORTANCE: This study addresses a critical challenge in drug susceptibility testing (DST): the discrepancies in DST results for isoniazid (INH) between the Bactec MGIT 960 system and the GenoType MTBDRplus assay. These discordant results significantly complicate treatment decisions, potentially leading to suboptimal patient outcomes. Using MIC assays and WGS on 53 clinical Mycobacterium tuberculosis strains, we provide valuable insights into the genetic basis of INH resistance. Our findings showed that only a small fraction of strains carried variants definitively linked to INH resistance, while a larger number harbored variants of uncertain significance across multiple genes, underscoring the complexity of INH resistance mechanisms. This study highlights the urgent need to refine our understanding of these "Group 3: uncertain significance" variants, as they appear to be a primary driver of the discrepancies. Additionally, this study emphasizes the importance of integrating advanced sequencing tools into DST to improve the accuracy of INH resistance detection.

Isoniazid

Characterization of Staphylococcus aureus isolates with decreased susceptibility to vancomycin and teicoplanin: isolation and purification of a constitutively produced protein associated with decreased susceptibility.

"Derivative isolates" with 4- to 8-fold and 8- to 16-fold increases in MICs of vancomycin and teicoplanin, respectively, were selected from 2 susceptible clinical isolates of Staphylococcus aureus by serial incubation in low-level vancomycin. A protein of approximately 39 kDa was demonstrable in the cytoplasmic fraction and occasionally in the membrane fraction by SDS-PAGE of both derivatives. This protein was purified by DEAE chromatography, preparative SDS-PAGE, and electroelution. Derivative bacteria were larger on transmission electron microscopy, had thicker cell walls, and had changes in colony morphology on solid media. Further evidence for cell wall reorganization included loss of phage and capsular typing, decreased susceptibility to lysostaphin/lysozyme killing, and changes in condition for detection of optimal coagulase activity. The mechanism of decreased susceptibility to glycopeptide antibiotics among S. aureus derivative isolates is uncertain. The production of the approximately 39-kDa cytoplasmic protein and cell wall reorganization may mediate changed affinity of glycopeptide-peptidoglycan binding or impairment of glycopeptide access to its cell wall target.

Bacterial Proteins

Effect of age of non-skin tissues on susceptibility of skin grafts to 7,12-dimethylbenz[alpha]anthracene (DMBA) carcinogenesis in BALB/c mice, and effect of age of skin graft on susceptibility of surrounding recipient skin to DMBA.

The influence of age-dependent alterations in non-skin tissues on chemical carcinogen-induced skin papilloma development was studied by treatment with 7,12-dimethylbenz[alpha]anthracene (DMBA) of 4-month-old skin grafts sewed onto 4- and 20-month-old syngeneic recipients. Skin of 4- and 20-month-old BALB/c female mice differed in susceptibility to DMBA carcinogenesis, but the 4-month-old grafts showed the same papilloma incidence independent of the age of the recipient mice. In a different experiment, the influence of age of skin grafts on papilloma development on DMBA-treated recipient skin was studied. Fourteen- and 26-month-old skin grafts were carried by 14-month-old recipients. Grafts of those two ages are known to differ in susceptibility to DMBA carcinogenesis, but no effect of the grafts on papilloma development on DMBA-treated recipients was detectable. It was concluded that certain age-dependent differences in skin susceptibility to chemical carcinogens are solely a reflection of alterations in the skin at the site of carcinogen treatment.

9,10-Dimethyl-1,2-benzanthracene

Resistance and susceptibility of mice to bacterial infection: course of listeriosis in resistant or susceptible mice.

Resistance and susceptibility to Listeria monocytogenes in mice was found to be related to (i) the innate ability of the nonimmune macrophages to kill or inhibit the growth of the organism during the first 24 to 48 h after infection, and (ii) the time of onset of acquired cell-mediated resistance. Resistant C57Bl/6 mice were 10 times more efficient than susceptible BALB/c mice at suppressing the early growth of Listeria in the liver. Furthermore, the onset of acquired immunity occurred 24 to 48 h earlier in C57Bl/6 than in BALB/c mice. Acquired immunity was measured by (i) fall in bacterial numbers in spleen and livers of infected mice (ii) adoptive transfer of immunity to normal mice by using spleen cells from infected mice, (iii) delayed-type hypersensitivity skin testing, and (iv) uptake of tritiated thymidine by lymphocytes in the spleen.

Animals

Succinate-DCPIP and NADH-fumarate oxidoreductases in fresh water snails susceptible and non susceptible to schistosoma infection.

The activities of succinate-DCPIP oxidoreductase (SO) and NADH-fumarate oxidoreductase (FR) were determined in tissue homogenate of Biomphalaria alexandrina and Bulinus truncatus, the snail vectors of Schistosomiasia. A parallel study was done on Lymnea truncatula snails which are not susceptible to Schistosoma infection. The Michaelis constant (Km) and maximum velocities (Vmax) for fumarate reduction and succinate oxidation by the tissue homogenates from the three species were determined. The results obtained showed that both susceptible species are aerobic and lactate is the sole end product of anaerobic glycolysis. Lymnea truncatula snails are facultative anaerobic producing succinate as a major end product in the glycolytic pathway.

Animals

Mammalian serum susceptibility of Serratia marcescens: detection of three human serum susceptibility categories.

A total of 65 strains of S. marcescens, including 36 currently employed O- and H-antigen reference strains, were examined for their susceptibility to the bactericidal activity of 80 vol % of fresh human serum. The majority of strains (57 = 87.6%) proved 'delayed serum-sensitive' (DSS); 4 strains (6.2%) were 'promptly serum-sensitive' (PSS), whereas 4 strains (6.2%) resisted (NSS) complement-mediated killing by human serum. Fresh sera obtained from 7 adult human volunteers yielded essentially identical kinetic killing data against representative NSS, DSS and PSS strains of S. marcescens. These observations were interpreted as further justification for the assignment of clinical isolates of S. marcescens to 3 categories of human serum susceptibility: DSS, PSS and NSS. Species differences among fresh sera from sheep, rabbits and guinea pigs, as contrasted with fresh human serum, were noted. In general, PSS strains of S. marcescens were killed in a slightly delayed fashion; DSS strains of S. marcescens were killed in an unpredictable kinetic pattern by sheep and rabbit sera, whereas fresh guinea pig serum entirely failed to kill selected DSS isolates of S. marcescens.

Adult

Resistance and susceptibility to the induction of rat adjuvant disease. Diverging susceptibility and severity achieved by selective breeding.

Rats selected for their ability to develop or resist adjuvant disease were used to establish 2 inbred lines of rat over 20 generations. A resistant line was rapidly established with almost 100% non-responsiveness by the sixth generation. A line showing 100% susceptibility was also established very rapidly but throughout the course of the breeding programme the severity continued to increase in intensity to a level considerably above that to be seen in strains normally considered to be high responders. At the thirteenth generation and beyond, the susceptible line showed a marked sex difference in the secondary lesions, females being more severely affected than the males. The 2 lines of rat were also tested for their ability to develop experimental allergic encephalomyelitis (EAE) in selected generations. There was no clear correlation between the 2 diseases although those animals developing the most severe adjuvant disease also had the most severe EAE.

Animals

Does the locus on chromosome 11 implicated in susceptibility to HLA-DR4 dependent type I diabetes mellitus also affect susceptibility to rheumatoid arthritis?

There is a polygenic component to rheumatoid arthritis (RA) in addition to the known association with HLA-DR4. It has previously been shown in another autoimmune disease (type I diabetes mellitus) that a gene on chromosome 11p can act with HLA-DR4 to enhance susceptibility (relative risk 5-6). It is therefore possible that this locus may also affect the development of RA. Genotype frequencies at this locus, defined by a dimorphic Fok 1 restriction site, were compared in 139 healthy controls and 213 patients with classical/definite RA. In contrast with diabetes there was no increase in genotypes lacking the Fok 1 site, either in the rheumatoid group overall (125/211 compared with 86/139 controls) or in the DR4 positive rheumatoid group (76/140 compared with controls). These results indicate that the interaction between DR4 and a locus on chromosome 11p is not common to all DR4 associated autoimmune diseases.

Amino Acid Sequence

The localization of influenza virus in the respiratory tract of ferrets: susceptible nasal mucosa cells produce and release more virus than susceptible lung cells.

Infectious virus production by ferret nasal mucosa and lung organ cultures has been monitored in both tissue pieces and medium over 24 h following inoculation with an Asian (H2N2) strain of influenza virus. Freshly prepared cultures of nasal mucosa produced approx. 10-fold more virus per cell than fresh lung cultures. Also the nasal mucosa cells liberated into the medium a greater proportion (mean 31%) of the total virus produced than did fresh lung (mean 6%). Maintenance of lung explants for 24 h prior to inoculation resulted in a 20- to 100-fold increase in the amount of virus released. However, total virus production by fresh and maintained lung was similar. Trypsin did not increase the infectivity of virus released from any of the cultures, indicating that the haemagglutinin in the virus particles was cleaved. Similar results were obtained with a Hong Kong (H3N2) virus strain. Hence, one factor operating in the lower susceptibility of the lung compared with the nasal mucosa in vivo may be a lower capacity of lung cells both to produce and release influenza virus.

Animals

Susceptibility of anaerobic bacteria to metronidazole, ornidazole, and tinidazole and routine susceptibility testing by standardized methods.

A total of 114 strains of anaerobic bacteria were examined for their susceptibility to metronidazole, ornidazole, and tinidazole by measuring the minimum inhibitory concentration (MIC) and minimum bactericidal concentration in different media. All strains, with the exception of the isolates of Propionibacterium acnes, were inhibited by 3.1 mug each and killed by 6.3 mug each of all three nitroimidazole compounds per ml. No significant differences in MIC values were found among metronidazole, ornidazole, and tinidazole. Only minor differences were detected by comparing MIC values obtained in brain heart infusion agar with and without sheep blood, brucella agar, and Mueller-Hinton agar (both containing blood). When the strains were tested by the modified broth-disk method proposed by the Anaerobe Laboratory of the Virginia Polytechnic Institute (VPI), there was good correlation with the MIC values (97.4% agreement for metronidazole and 94.7% for ornidazole and tinidazole). For routine testing, use of a 30-mug-class disk of either nitroimidazole derivative is proposed for the broth-disk method, resulting in a final concentration of 6 mug/ml in the test tubes, a concentration easily attainable in body fluids. In contrast to the broth-disk method, there was very poor correlation between inhibition zone diameters by the standardized VPI agar diffusion test and MIC values.

Anaerobiosis

[Synthetic medium for testing of susceptibility. II. Susceptibility of bacteria to sulphonamides (author's transl)].

A comparative study of 9 media for testing the susceptibility of bacteria to 7 sulphonamides has been carried out. The evaluated culture-media included 4 "Mueller Hinton" media (MH), 4 others based on the formula of the "Diagnostic Sensitivity Test" agar base (D.S.T.) and a "Synthetic Medium" (SM) which has been developed in our laboratory. MH and D.S.T. culture-media have been supplied by 4 manufacturing companies in Europe and USA. A total number of 21 bacterial cultures have been employed. The included species belong to the gram negative genera Citrobacter, Escherichia, Klebsiella, Proteus, Salmonella, Serratia and Shigella, and to the gram positive genus Streptococcus. Sulphafurazole, sulphasomidine, sulphamethoxazole, sulphamethoxypyridazine, sulphadimethoxine, sulphaphenazole and sulphamethoxydiazine have been investigated. SM proved to be clearly superior to the other 8 culture-media mentioned above. Just one MH afforded good results. The 3 other MH and 2 D.S.T. media were not quite satisfactory. The two remaining D.S.T. media were unsuitable. Moreover, the activity of the tested sulphonamides was not equally high against the concerned bacteria. The author discussed the significance of the inoculum-size and of competitive and noncompetitive sulphonamide-antagonists in commercial culture-media and the difficulties they raise by the interpretation of the results.

Bacteria

The epidemic in a closed population with all susceptibles equally vulnerable; some results for large susceptible populations and small initial infections.

Kendall's (1956) approach to the 'general' epidemic is generalized by dropping the assumptions of constant infectivity and random recovery or death of ill individuals. A great deal of attention is paid to the biological background and the heuristics of the model formulation. Some new results are: (1) the derivation of Kermack's and McKendrick's integral equation from what seems to be the most general set of assumptions in section 2.2, (2) the use of Kermack's and McKendrick's final value equation to arrive at a finite time version of the threshold theorem for the general case, comparable to the for the case of only one Markovian state of illness in section 2.5, (3) the analysis of the behaviour of the solutions of the integral equation when the starting infection approaches zero in section 2.7, (4) the derivation of the probability structure of a general branching process, after conditioning on extinction in section 3.6, (5) the statement of the generalized versions of Kendall's ideas in the form of precise limit conjectures in section 4, (6) the derivation of a closed expression for the limit epidemic resulting from (3) in appendix 4.

Disease Outbreaks