Mold Extracts Research Articles

Double-blind, placebo-controlled rush immunotherapy with a standardized Alternaria extract

Specific immunotherapy is ineffective with unstandardized mold extracts. A double-blind, placebo-controlled study was performed in 24 patients (5 to 56 years of age) only allergic to Alternaria. The extract was standardized by isoelectric focusing, crossed immunoelectrophoresis, crossed radioimmunoelectrophoresis, RAST inhibition, and skin tests and contained allergen Alternaria major allergen a I and antigen B. Thirteen patients received the active treatment, and 11 received the placebo. Immunotherapy was started by a 2-day rush protocol; maintenance injections were administered for 1 year. The patient's self-evaluation of the treatment was significantly ( p < 0.001) lower in the placebo-treated group. Global symptom-medication scores, including asthma and rhinoconjunctivitis, were significantly ( p < 0.005) lower in the actively treated group. Nasal challenges with Alternaria extract were performed before immunotherapy and after 1 year of treatment. There was no difference in the placebo-treated group and a significantly ( p < 0.01) increased mean provocative dose in the actively treated group. Skin tests were significantly reduced in the actively treated group. Specific IgG increased significantly in the actively treated group and were stable in the placebo-treated group. There was a significant correlation between nasal challenges and nasal symptom-medication scores ( p < 0.03) or the patient's self-evaluation of efficacy ( p < 0.05). This study demonstrated that patients only sensitized to Alternaria benefit from specific immunotherapy with a standardized Alternaria extract.

Allergens

The environment contains a huge array of allergens, primarily proteins or glycoproteins, to which the atopic individual may become sensitized. In situations of high exposure and highly reactive substances, even nonatopic individuals may develop allergic antibodies. The individual’s response depends on the underlying immune responsiveness, the dosage, frequency, route, and possibly the chemical nature of the allergen. Most allergenic sources contain multiple allergenic determinants, as demonstrated by characterization of pollen and mold extracts. Major allergens are acidic proteins with weights in the 20,000 to 40,000 dalton range, and pls in the 4 to 6 range. Minor allergens may be larger or smaller and either acidic or basic. These conclusions are based on the isolation of a relatively small number of allergens. Cross-reactivity data, especially with the pollens, are useful in the rational formulation of immunotherapy extracts. Considering the wealth of allergenic pollens, information available to date is very scanty. Data on molds, epidermals, and mites are even more preliminary or conflicting. As further research proceeds on purifying and characterizing allergens, a number of these questions and conflicts will be resolved.

The Nature of Allergens

The environment contains a huge array of allergens, primarily proteins or glycoproteins, to which the atopic person may become sensitized. In situations of high exposure and highly reactive substances, even nonatopic persons may develop allergic antibodies. The individual’s response depends on the underlying immune responsiveness, the dosage, frequency, route, and possibly the chemical nature of the allergen. Most allergenic sources contain multiple allergenic determinants, as demonstrated by characterization of pollen and mold extracts. Major allergens are acidic proteins with weights in the 20,000 to 40,000 dalton range, and isoelectric points in the pH 4 to 6 range. Minor allergens may be larger or smaller and either acidic or basic. These conclusions are based on the isolation of a relatively small number of allergens. Cross-reactivity data, especially with pollens, are useful in the rational formulation of immunotherapy extracts. But, considering the wealth of allergenic pollens, the information available to date is very scanty. The data on molds, epidermals, and mites are even more preliminary or conflicting. As further research proceeds on purifying and characterizing allergens, a number of these questions and conflicts will be resolved.

Formation equation of halogenated organic compounds when water is chlorinated

Various halogenated organic compounds are formed by chlorination of water. In this study, formation of organic compounds halogenated from a reagent humic acid and extract of a leaf mold were examined under various conditions. The following overall formation equation was obtained from empirical data under the practical wide range when free chlorine remained. [ TOX]=k TOX [ TOC][ Cl 2] α ot β. Here, [TOX] is the concentration of total organic halogen after t h in units of mg chlorine per liter; [TOC] and [Cl 2] o are concentrations of total organic carbon and dosed chlorine in units of mg per liter; k TOX is the rate constant and α and β are parameters. From the values of k TOX, α and β, the character of organic substances i.e. precursor of halogenated organic compounds, in water can be evaluated. The values k TOX, α and β for humic acid are 0.053, 0.28 and 0.13, and the values for extract of the leaf mold are 0.032, 0.36 and 0.15, respectively. The activation energies are 10 kJ mol −1 and 11 kJ mol −1 for the reactions of humic acid and leaf mold extract, respectively.

Basidiomycete mycelia and spore-allergen extracts: Skin test reactivity in adults with symptoms of respiratory allergy

One hundred fifty adults, with respiratory-allergic disease, and 14 control subjects, without symptoms of respiratory allergy, were skin prick tested with 16 common inhalant allergens, 12 extracts of mycelia from Basidiomycetes grown in vitro, and/or 10 to 15 basidiospore extracts. Eighty-three subjects (58%) had positive skin tests to two or more of the common inhalant allergens. Twenty-seven percent of the study subjects had positive skin reactions to one or more of the Basidiomycete mycelia extracts, and 32% demonstrated positive skin reactions to one or more basidiospore extracts. None of the 14 control subjects had positive skin reactivity to basidiospore extracts. Skin prick reactivity of study subjects to 15 different basidiospore extracts ranged from 5% for Cantharellus cibarius to 17% for Scleroderma sp. The prevalence of skin test reactivity to basidiospores did not differ significantly from the reactivity to commercial mold extracts of several common species of the Fungi Imperfecti (6% for Cladosporium herbarum or Penicillium notatum to 13% for Alternaria tenuis). These results demonstrate that a significant number of individuals reporting symptoms of respiratory allergy have skin test reactivity to basidiospores and suggest that these spores are important fungal aeroallergens in the New Orleans environment.

Production of glutathione peroxidase by Mucor hiemalis.

The distribution of glutathione peroxidase was investigated among microorganisms. Glutathione peroxidase was found in the cell-free extracts of molds belonging to the genus Mucor, and Mucor hiemalis showed the highest enzyme activity. The glutathione peroxidase production by M. hiemalis was almost completely associated with the mycelial growth. The addition of oxidants such as hydroperoxides and antioxidants such as glutathione and α-tocopherol did not significantly affect the enzyme production.

Studies on microbial glycerol dehydrogenase. Part I. Formation of glycerol dehydrogenase by microorganisms.

The distribution of glycerol dehydrogenase activity was studied with cell-free extracts of bacteria, yeasts, molds and actinomycetes. High activity was found in 4 strains of bacteria and in 3 strains of molds. The enzymes of bacteria were dependent on NAD+ and those of molds were dependent on NADP+. An isolated gram-positive bacterium, which showed the high activity, was identified as Cellulomonas sp. NT3060. The total and specific activities were associated with growth of this strain and reached the maximum at the early stationary phase. Significant high level activity was detected in cell-free extracts from glycerol and glucose media.

Human Malignant Melanoma: Assay of Tumor Associated Antigens by Use of Rabbit Antisera

The usefulness of rabbit antisera for the characterization of tumor associated antigens of human melanoma was studied. Membranes of human melanoma tissue were extracted by 2M potassium iodide and sonication. Antisera were raised in rabbits against these extracts as well as against melanoma membranes and cultured melanoma cells. Antisera were absorbed with normal human material. Two of 14 absorbed antisera were specific for tumor associated antigens: they reacted with melanoma extracts (counterimmunoelectrophoresis) and melanoma cells (immune adherence) but not with 20 extracts of normal tissue, erythrocytes, lymphocytes, bacteria and molds (counterimmunoelectrophoresis), not with human fibroblasts (immune adherence), not with human erythrocytes with known blood group antigens (hemagglutination) and normal human skin (immunofluorescence). Rabbit antisera did not contain antibodies against carcinoembryonic antigen and alpha-i fetoprotein (radioimmunoassay). Reactivity with tumor associated antigens could be directly demonstrated by two-dimensional immunoelectrophoresis. Two types of melanoma associated antigens could be discriminated: the first type with restricted specificity could be demonstrated by an antiserum that reacted with the extract used to raise it and in addition had a weak reactivity with 1 of 7 other extracts. Broader cross reactivity is characteristic for the second type: the corresponding antiserum reacted with 6 of 8 melanoma extracts.

Production and purification of choline oxidase from Cylindrocarpon didymum M-1.

The distribution of choline oxidase activity was studied with cell-free extracts of yeasts, molds and actinomycetes. A fungus which was identified as Cylindrocarpon didymum M–1 showed the highest activity. The enzyme was purified from the cell-free extract of C. didymum M–1 by a procedure involving ammonium sulfate fractionation and DEAE-cellulose, hydroxyl-apatite and Sephadex G–150 column chromatographies. The enzyme preparation was homogeneous when subjected to disc gel electrophoresis and ultracentrifugation. Sedimentation velocity yielded a value of . The enzyme showed a typical flavoprotein spectrum of absorption maxima at 276,370 and 454 nm.

Membrane associated antigens of human malignant melanoma V: Serological typing of cell lines using antisera from nonhuman primates

Antisera against various melanoma cell lines were raised in nonhuman primates (Cercopithecus aethiop.). After exhaustive absorption with AB Rh + red blood cells and pooled platelets from about 200 donors the sera were still reactive to various degrees in the microimmune adherence test with other melanoma lines, with embryonic fibroblasts, and with non-melanoma lines. As proven by absorption experiments, the main-specificity of the antisera was not directed against components of the fetal calf serum used for cell culture or against mycoplasma grown from commercial fetal calf serum. In addition, no cross-reactivity was observed with Bacillus Calmette-Gerin, and in blocking experiments no reactivity against extracts of common bacterial antigens or mixed molds was detected. Absorption with embryonic fibroblasts or embryonic tissue showed that the reactivity of most antisera was directed against melanoma-associated antigens expressed also on fetal tissue. It was not possible to determine whether the remaining reactivity on some cell lines was melanoma-specific or directed against fetal antigens not contained in the fetal material used for absorption. Cross-absorption of antisera with other melanoma cells revealed that various cell lines express different patterns of tumor-associated antigens with no, or only partial, overlap. The cross-absorption experiments made it possible to type the cell lines according to their surface antigens and arrange the cell lines in order according to the degree of mutual antigenic relationship.

Photoreceptor pigment that induces differentiation in the slime mold Physarum polycephalum.

An extract of small molecules (molecular weight less than 500) of the slime mold Physarum polycephalum undergoes a shift in ultraviolet-visible absorption spectrum upon illumination. This illumination also confers on the extract the ability to induce sporulation when injected into a starved, unilluminated slime mold. The spectral shift and appearance of the sporulation-inducing activity both occur regardless of whether the illumination is carried out on an intact slime mold or on the plasmodium-free extract itself. Thin-layer chromatography resolves the slime mold extract into four major visible fractions. One of these has high sporulation-inducing activity after illumination in vitro.

Brachial Plexus Neuropathy

Two patients had acute, self-limiting, unilateral brachial plexus neuropathy in association with desensitizing subcutaneous injections of extracts of dusts and molds. Diminution of pain and return of function of the affected limb followed a short course of prednisone therapy, although a beneficial effect of the steroid cannot be considered proved. There is a large variety of antigenic substances already known to produce neuropathy following intramuscular injection; desensitizing antiallergy injections should be added to this list.

Brachial plexus neuropathy. Association with desensitizing antiallergy injections

Two patients had acute, self-limiting, unilateral brachial plexus neuropathy in association with desensitizing subcutaneous injections of extracts of dusts and molds. Diminution of pain and return of function of the affected limb followed a short course of prednisone therapy, although a beneficial effect of the steroid cannot be considered proved. There is a large variety of antigenic substances already known to produce neuropathy following intramuscular injection; desensitizing antiallergy injections should be added to this list.

Guillain-Barré syndrome.

To the Editor.— The letter by Anderson (220:127, 1972) reporting the Guillain-Barre syndrome as a complication of allergic immunotherapy brought to mind a young man under my care during neurology residency who developed a hemiparesis following an influenza vaccine injection. Within one year he developed other episodes typical of multiple sclerosis. In reviewing the literature for information about allergy and central nervous system disorders, it became apparent that the Landry-Guillain-Barre syndrome had been a frequent expression of such reactions. E. Uehlinger ( Schweiz Med Wochenschr 87:813, 1957) has reported Landry's paralysis after poliomyelitis vaccination. It is a well-known complication of antirabies vaccine, and, indeed, almost every known prophylactic inoculation: Miller and Stanton ( Quart J Med 23:1, 1954). Although I know of no case following a pollen, dust, mold extract injection, I share Dr. Anderson's concern about a possible causal relationship after learning about the vagaries of foreign protein reactions and the

Structural differences in vinblastine-induced aggregates of acidic proteins

High speed supernatants of pig brain, rabbit brain, the slime mould Physarum polycephalum as well as purified rabbit and myxomycete actin were incubated with vinblastine sulfate (1 × 10 −3 M). Thin sections of the resulting precipitates revealed different ultrastructures when examined by electron microscopy. While precipitates derived from slime mould extracts showed exclusively amorphous, densely stained aggregates which looked similar to those obtained with purified actin, the precipitates from brain extracts were heterogeneous. They contained hexagonal and helical structures in addition to amorphous material. The ordered aggregates are thought to be composed of microtubular protein, while the amorphous material probably contains actin, as well as possibly other proteins.

Leloir's pathway in Polyporus circinatus.

Polyporus circinatus grows in galactose as sole carbon source. The enzymes involved in galactose metabolism were investigated in the mold extracts. Evidences for the enzymes activities were obtained by the analysis of products through tracing techniques and paper chromatography. Galactokinase, Gal-1-P uridyl transferase, UDPGal-4-epimerase, and UDPGal pyrophosphorylase could be detected.

Delayed Reaction to Aqueous Hyposensitization Material

To the Editor.— Delayed local reaction to hyposensitization emulsion extracts have been a major drawback in their use. 1 However, delayed local reactions rarely occur with aqueous hyposensitization extracts. 2,3 We report a case of what seems to be a delayed local reaction to aqueous hyposensitization extract. Report of a Case.— A 17-year-old Negro girl showed on the arm starting in September 1967. Three months later, the lumps began to drain serous watery material. The patient had developed seasonal asthma (June to September) at 11 years of age. Thereafter, she received epinephrine in aqueous or oily suspension in the deltoid area. The last epinephrine in oil injection was received in August of 1966. Skin tests revealed positive reactions to house dust, trees, grass and weed pollens, and mold extracts. Subcutaneous injections of aqueous preparations of these were administered from Aug 18, 1966 to Sept 14, 1967. The maintenance dose

Mold allergy: Fractionation of allergens by Sephadex G-200 gel filtration

The fractionation of mold extracts by Sephadex G-200 revealed, with some molds, three peaks absorbing light at 280 mμ. Skin reactivity and precipitation were localized in the small and middle peaks, while the larger peak contained the mold pigments and ninhydrin staining components. In some molds the middle peak was absent, but in these cases the skin and precipitation activities were found in the first fractions of the larger peak. Individual variations were observed among the patients in their skin reactivity to the various molds. Some patients reacted to both the small and middle peaks, but others reacted to only one of them. It is evident, therefore, that mold allergens can be divided into two groups on the basis of molecular size. In the case of Alternaria, the thermostable antigens were all concentrated in the small peak; thus, they have a larger molecular size than the thermolabile antigens which came out in the middle peak.

Cell wall synthesis in Dictyostelium discoideum: A model system for the synthesis of alkali-insoluble cell wall glycogen during differentiation

An enzyme present in the 100,000 g pellet fraction of slime mold extracts has been shown to catalyze the synthesis of alkali-insoluble cell wall glycogen from UDPG- 14C. This reaction is completely primer-dependent, in contrast to the system in which cell wall preparations are used as a source of both enzyme and primer. Four lines of evidence suggest that one enzyme in the pellet fraction catalyzes the synthesis of both soluble glycogen and insoluble cell wall glycogen. The more insoluble the primer used as acceptor for this enzyme, the less dependence there is on G-6-P. In the presence of pellet enzyme, with glycogen as primer, the K m for UDPG is 4.2 × 10 −4 m in the presence of G-6-P and 4.7 × 10 −3 m in the absence of G-6-P; with cell wall as primer, the K m is 4.6 × 10 −4 m in the presence of G-6-P and 8.3 × 10 −4 m in the absence of G-6-P. When cell wall is the source of both enzyme and primer, the K m is about 1.5 × 10 −3 m in the presence or absence of G-6-P.

Toxic fungi isolated from tall fescue.

Extracts of cold-grown common molds isolated from tall fescue forage produce toxic effects on rabbit skin and hemorrhagic death in mice. A mixture ofCladosporium cladosporioides andFusarium nivale produced ruminal atony in a sheep. The clinical findings in the sheep show some similarity to those of cattle on toxic fescue pastures. Further work to determine the role of fungal toxins in the etiology of fescue foot in cattle is underway.