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PubMed1989

Tissue cooperation in a proteolytic cascade activating human interstitial collagenase.

We present a cascade of proteolytic events catalyzed by the proteases secreted by cultured keratinocytes and fibroblasts that results in the activation of interstitial procollagenase. Cultured human skin fibroblasts constitutively secrete interstitial collagenase and stromelysin as proenzymes. In contrast, interstitial collagenase found in serum-free skin organ culture conditioned medium is activated. Cocultivation of the major cellular components of skin organ culture, dermal fibroblasts and epidermal keratinocytes, induces activation of interstitial procollagenase and prostromelysin in the presence of plasminogen. This activation occurs through a urokinase-dependent pathway where added keratinocytes secrete the plasminogen activator urokinase, which converts plasminogen into plasmin. Plasmin is capable of activating purified procollagenase and prostromelysin. Plasmin-dependent activation of procollagenase generates an enzyme species, by amino-terminal processing, identical to those generated by limited proteolysis with trypsin or treatment with organomercurial compounds. Catalytic amounts of activated stromelysin can in turn convert plasmin- or trypsin-activated collagenase into a fully active enzyme by removal of approximately 15 amino acid residues from the carboxyl end of the enzyme. This results in a 5- to 8-fold increase in collagenase specific activity that is due to its proteolytic cleavage and not to the presence of the activator stromelysin. Stromelysin alone in both pro- and activated forms is not capable of efficient activation of human fibroblast interstitial procollagenase.

Cells, Cultured↗

PubMed2006

CD4 T cell control primary measles virus infection of the CNS: regulation is dependent on combined activity with either CD8 T cells or with B cells: CD4, CD8 or B cells alone are ineffective.

Measles virus (MV), one of the most infectious of human pathogens, still infects over 30 million humans and causes over 500,000 deaths each year [Griffin, D., 2001. Measles virus. In: Fields, B., Knipe, D., Howley, P. (Eds.), Fields Virology. Lippincott-Raven, Philadelphia, pp. 1401-1442; ]. Death is primarily due to secondary microbial infections associated with the immunosuppression caused by MV. Studies of humans with genetic or acquired deficiencies of either the humoral or cellular arm of the immune system, and rodent models have implicated T cells in the control of the ongoing MV infection but the precise role and activities of the specific T cell subset or the molecules they produce is not clear. Using a transgenic mouse model in conjunction with depletion and reconstitution of individual B and T cell subsets alone or in combination, we show that neither CD4, CD8 nor B cells per se control acute MV infection. However, combinations of either CD4 T cells and B cells, or of CD4 and CD8 T cells are essential but CD8 T with B cells are ineffective. Interferon-gamma and neutralizing antibodies, but neither perforin nor TNF-alpha alone are associated with clearance of MV infection. TNF-alpha combined with interferon-gamma is more effective in protection than interferon alone. Further, the lack of an interferon-gamma response leads to persistence of MV.

Adoptive Transfer↗

PubMed2004

Biophysical and kinetic analysis of wild-type and site-directed mutants of the isolated and native dehydroquinate synthase domain of the AROM protein.

Dehydroquinate synthase (DHQS) is the N-terminal domain of the pentafunctional AROM protein that catalyses steps 2 to 7 in the shikimate pathway in microbial eukaryotes. DHQS converts 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) to dehydroquinate in a reaction that includes alcohol oxidation, phosphate beta-elimination, carbonyl reduction, ring opening, and intramolecular aldol condensation. Kinetic analysis of the isolated DHQS domains with the AROM protein showed that for the substrate DAHP the difference in Km is less than a factor of 3, that the turnover numbers differed by 24%, and that the Km for NAD+ differs by a factor of 3. Isothermal titration calorimetry revealed that a second (inhibitory) site for divalent metal binding has an approximately 4000-fold increase in KD compared to the catalytic binding site. Inhibitor studies have suggested the enzyme could act as a simple oxidoreductase with several of the reactions occurring spontaneously, whereas structural studies have implied that DHQS participates in all steps of the reaction. Analysis of site-directed mutants experimentally test and support this latter hypothesis. Differential scanning calorimetry, circular dichroism spectroscopy, and molecular exclusion chromatography demonstrate that the mutant DHQS retain their secondary and quaternary structures and their ligand binding capacity. R130K has a 135-fold reduction in specific activity with DAHP and a greater than 1100-fold decrease in the kcat/Km ratio, whereas R130A is inactive.

Alcohol Oxidoreductases↗

PubMed1981

[A computerized information system of bacillary tuberculosis in the Czech Socialist Republic (author's transl)].

The automated information system for bacillary tuberculosis is based on the registration of all positive mycobacterial examinations in the 43 tuberculosis laboratories in CSR. Data on the patient, the health service reporting, the kind of the material examined, and the actual microbiological examination including the identification of species and sensibility to antituberculotics are surrendered to the computer by means of a punched card or tape. On the base of the entrance information all sources are collected in the computer the data input being checked in regard to accuracy of the parts and logical relationship to the various variables. All sources of information of the data bank, made actual concerning the mycobacterial examinations are available to the programme. Another programme is supplementing the information on tests for identification and antituberculotic sensibility of the mycobacteria. With the aid of other programmes regular monthly summaries are done by the data bank for the cooperating laboratories and the specialists in the districts and countries. Finally another programme answers the evaluation of the annual reports of the mycobacterial investigations and reports on the epidemiological data of the diseased persons. In 1977 13,909 notifications of tuberculosis findings were analysed, coming from 3,782 persons. This analysis made evident the following conclusions reflecting the recent situation of mycobacteriologic diagnostics and the significance of the tuberculosis problem in communities with a low tuberculosis prevalence: a) The prevalence of bacillary tuberculosis in CSR (37/100,000) is ranging in the various districts between 27 and 44/100,000. The prevalence of persons with proof of mycobacteria already by smear amounts to 10/100,000 inhabitants. b) The highest portion of bacillary tuberculosis comes from persons with symptoms (45%), it amounts to 17% in persons with active tuberculosis already in dispensary care, to 15% in persons with fibrotic lesions and 14% in recidives. After a five year experience with the computerized information system of bacillary tuberculosis the authors recommended to centralize and to combine it with the central state tuberculosis register in order to utilize the data and evaluated in the level of the districts and countries as well for the management of the tuberculosis control programme in the level of the whole state.

Czechoslovakia↗

PubMed2002

DNA polymerase gene sequences indicate western and forest tent caterpillar viruses form a new taxonomic group within baculoviruses.

Baculoviruses infect larval lepidopterans, and thus have potential value as microbial controls of agricultural and forest pests. Understanding their genetic relatedness and host specificity is relevant to the risk assessment of viral insecticides if non-target impacts are to be avoided. DNA polymerase gene sequences have been demonstrated to be useful for inferring genetic relatedness among dsDNA viruses. We have adopted this approach to examine the relatedness among natural isolates of two uncharacterized caterpillar-infecting baculoviruses, Malacosoma californicum pluviale nucleopolyhedrovirus (McplMNPV) and Malacosoma disstria nucleopolyhedrovirus (MadiMNPV), which infect two closely related host species with little to no cross-infectivity. We designed two degenerate primers (BVP1 and BVP2) based on protein motifs conserved among baculoviruses. McplMNPV and MadiMNPV viral DNA was obtained from naturally infected caterpillars collected from geographically distinct sites in the Southern Gulf Islands and Prince George regions of British Columbia, Canada. Sequencing of 0.9 kb PCR amplicons from six McplMNPV and six MadiMNPV isolates obtained from a total of eight sites, revealed very low nucleotide variation among McplMNPV isolates (99.2-100% nucleotide identity) and among MadiMNPV isolates (98.9-100% nucleotide identity). Greater nucleotide variation was observed between viral isolates from the two different caterpillar species (only 84.7-86.1% nucleotide identity). Both maximum parsimony and maximum likelihood phylogenetic analyses support placement of McplMNPV and MadiMNPV in a clade that is distinct from other groups of baculoviruses.

Amino Acid Sequence↗

PubMed2005

Association of minimum inhibitory concentration cluster patterns with dairy management practices for environmental bacteria isolated from bulk tank milk.

Environmental bacteria have emerged over the past few years to become significant causes of mastitis. Bacteria in this group are often reported by practicing veterinarians to be increasingly resistant to intramammary therapy and responsible for elevated bulk tank somatic cell counts. The purpose of this study was to determine the extent of association of the minimum inhibitory concentrations for selected antimicrobial agents with environmental bacteria isolated from bulk tank milk on California dairies and their housing facilities, husbandry practices, and antimicrobic-use strategies. Bulk tank milk samples were collected from 2 dairy cooperatives that had their milk cultured at the Milk Quality Laboratory, University of California Davis, Veterinary Medicine Teaching and Research Center in Tulare, CA. Samples were collected from July 2001 through March 2002 on 88 d; and 404 environmental bacteria isolated from 93 dairies were found. Minimum inhibitory concentrations were determined on 337 of the isolates for 10 antimicrobial agents. Cluster analysis was performed on the minimum inhibitory concentration values for each organism, and 4 antimicrobial clusters with varying degrees of resistance were found.A 69-question survey questionnaire was completed on-farm for 49 of the 73 dairies that had at least 3 environmental bacterial isolates. The questionnaire sought information on housing facilities, milking management, mastitis prevention, antimicrobial usage strategies, and owner/veterinary involvement in disease control and prevention. Multinomial logistic regression analysis found significant associations between the antimicrobial agent-resistance cluster groups and some of the housing and bedding practices, failure to dry udders before milking, and antimicrobial treatment of nonmastitis conditions. No association was noted for antimicrobial agent treatment of mastitis and the resistance cluster patterns.

Animals↗

PubMed1995

[The bactericidal effects of an amphoteric surfactant with ethanol against Mycobacterium tuberculosis].

Bactericidal activities of an amphoteric surfactant (12 w/v% Alkyl diaminoethylglycine hydrochloride, 3 w/v% Alkyl dietylenetriaminoglycole hydrochloride, Nissan Anon #300, Inui Shouji Co., ADG) against Mycobacterium tuberculosis were examined independently or in cooperation with ethanol. The results obtained in the study are as follows: 1. In the treatment with the surfactant diluted with distilled water in one hundredth, the numbers of tubercle bacilli which survived were 500 cfu/0.1 ml after one minute-treatment, and 25 cfu/0.1 ml after ten minutes-treatment. On the other hand, the surfactant containing 23 v/v% ethanol decreased the bacterial numbers to seven cfu/0.1 ml after one minute and less than one cfu/0.1 ml after ten minutes. The numbers of the survived bacilli treated with the surfactant containing 20 v/v% ethanol were 500 cfu/0.1 ml after one minute-treatment, and less than one cfu/0.1 ml after ten minutes though 20 v/v% ethanol alone was ineffective in ten minutes-treatment. 2. The bactericidal activities of the surfactant against tubercle bacilli in sputum samples in which 1.65 x 10(4) cfu/0.1 ml of the organism were contained were also investigated. The numbers of tubercle bacilli which survived were 25 cfu/0.1 ml after one minute-treatment with the surfactant in one fiftieth, two cfu/0.1 ml after five minutes-treatment and one cfu/0.1 ml after ten minutes-treatment. The numbers of the organism treated with the surfactant containing 23 v/v% ethanol were less than one cfu/0.1 ml after five minutes. 3.(ABSTRACT TRUNCATED AT 250 WORDS)

Drug Resistance, Microbial↗

PubMed1988

Control of diarrhoeal diseases.

During its early years, WHO was primarily concerned with epidemic control, standardization and quality control of vaccines, and administration of the International Sanitary Regulations. Among the acute enteric infections, cholera received the greatest attention because of its propensity for rapid epidemic spread. When the seventh pandemic of cholera began in 1961, WHO responded with a greatly expanded programme of activities which included cooperation with countries in training and control efforts, and research on treatment and prevention, especially vaccine development. At the same time, numerous training courses were held on enteric infections and a series of controlled field trials established the degree of protection conferred by existing typhoid and paratyphoid vaccines. In 1970, when the cholera pandemic spread to Africa, the emergency assistance programme was reactivated, with increasing attention to the provision of appropriate treatment, especially oral rehydration therapy, rather than the supply of ineffective vaccines. The requirement of cholera vaccination for international travel in the International Health Regulations was abolished in 1973 by the World Health Assembly. Another public health problem of importance during the 1970s was the increase in antibiotic resistance of enteric bacteria, especially Shigella dysenteriae 1 and Salmonella typhi, first in Central America and Mexico and later in Asia. There was a notable acceleration in research on diarrhoeal disease and especially on cholera during this period, with the discovery of several new viral and bacterial agents of diarrhoea, advances in knowledge of intestinal immunity indicating that better protection might be achieved with oral vaccines, and the demonstration of the effectiveness of a single formulation of oral rehydration salts (ORS) in the treatment of all diarrhoeas including cholera.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholera↗

PubMedDate not supplied

[Late results of strictly supervised treatment of patients with newly detected pulmonary tuberculosis].

The late results are analized, of a strictly supervised treatment of 160 patients with pulmonary tuberculosis, of the secondary bacilliferic type, recently discovered, between July 1974 and December 1976. The criterium for assessing the results was the bacteriologic examination. After the first year 144 of the patients became negative (90%), while 16 remained positive (10%). Prolonged treatment, up to 2--3 years also led to negativation of these causes, with the only exception of a single patient that did not cooperate, and who became chronically ill. After one-and-a-half year 2 patients again became positive, of which one died with fulminating hemophytysis, and the second one in cardio-respiratory failure. Another 6 patients became positive after two years, while two other were positive at three years and one each after four, respectively five years. Two are still under treatment, while the remaining patients were all negative for at least two years. At the end of the observation period 155 of the initially 160 bacilliferic patients were considered as solved, and do not raise anti-epidemic problems. It is shown in conclusion that the modern anti-tuberculous treatment can lead to recovery of the majority of patients. Those who are still bacille carriers, or who recidivate, can also be solved by continued treatment for a period of up to 2--3 years. The patients with multiple deficiencies have increased risks to develop relcidives, and they should be followed with perseverance for a long time. The recommended therapeutic regimens are not significantly different from the viewpoint of their efficiency, provided they are administered in a strictly supervised manner. The reactivity of the organism probably has an important role in the efficiency of the modern anti-tuberculous treatment.

Adolescent↗

PubMed1994

Synergic inhibitory activity of amphotericin-B and gamma interferon against intracellular Cryptococcus neoformans in murine macrophages.

Cryptococcus neoformans is responsible for pulmonary and meningal infections in HIV patients. The lack of effective cellular cooperation caused by the low level of CD4+ cells, and the resistance of C. neoformans to phagocytosis allows growth and persistence of the yeast in the host. We describe here an in-vitro model of intracellular replication of C. neoformans inside J774-A.1 macrophages, and the determination of the intracellular antifungal activity of amphotericin B and fluconazole alone or in association with IFN-gamma. The maximum inhibitory effect was observed with one MIC of amphotericin B and 100 or 1000 IU/mL of IFN-gamma. amphotericin B alone (at 1 x MIC), or either 1 x or 50 x MIC of fluconazole in normal or IFN-gamma activated macrophages, did not eradicate the ingested yeast. A potential underlying mechanism of the synergy of amphotericin B in IFN-gamma primed macrophages was investigated by measurement of nitrite level and by use of the NO synthase competitive inhibitor, NG-monomethyl L-arginine (NMMA). One MIC of amphotericin B was able to activate the synthesis of nitrogen reactive intermediates in IFN gamma-primed macrophages. NMMA treated infected macrophages responded less well to IFN-gamma priming, resulting in a moderate inhibition in subsequent amphotericin B exposure.

Amphotericin B↗

PubMed1977

Mechanism of adjuvant activity of dental plaque: in vitro activation of residual helper T-cell precursors in T-cell-deficient murine spleen cell cultures.

The immunoenhancing activity of a water-soluble extract of dental plaque (DP), which contains a mixture of microbial antigens, has been investigated. DP was tested for its capacity to augment the in vitro antibody-forming cell (AFC) response to sheep erythrocyte by adherent spleen cells from thymectomized, lethally irradiated, and bone marrow-transplanted (TxB) mice. Although DP was found to induce a small polyclonal AFC response, most of the increase in AFC induced by DP was antigen dependent. The latter enhancing effect is an indicator of the adjuvanticity of DP. This adjuvant activity of DP was T-cell-dependent, since removal of the residual prethymic and/or thymic-derived lymphocytes (T-cells) by anti-T-cell serum (anti-theta) and guinea pig complement abrogated the capacity of DP to augment the in vitro AFC response. This view was further supported by the synergistic restorative effect obtained by culturing anti-theta-treated adherent spleen cells with both DP and a population of unactivated T-cells that by themselves were unable to significantly enhance AFC responsiveness. Moreover, DP was found to be mitogenic for thymocytes. The cumulative results suggest that the adjuvant activity of DP is dependent on both the T- and B-cell-activating components present in DP.

Adjuvants, Immunologic↗

PubMed1993

Zidovudine resistance, syncytium-inducing phenotype, and HIV disease progression in a case-control study. The VA Cooperative Study Group.

A case-control study of patients with progressive (cases) or nonprogressive (controls) disease was designed to determine the association among disease progression, zidovudine sensitivity, and syncytium-inducing phenotype. Viral isolates were screened for sensitivity to zidovudine using a peripheral blood mononuclear cell-based assay and for syncytium-inducing (SI) phenotype in MT2 cell culture. Thirty-four patients, whose disease progressed to AIDS or whose CD4 cell numbers fell < 200 cells/mm3, were matched with 34 patients whose conditions had not progressed or whose CD4 cell numbers remained > 200 cells/mm3. Virus was successfully cultured from both the progressor and the nonprogressor in 17 of these 34 matched case-control pairs. In six of the 17 pairs, virus isolated from the progressor had an IC50 (50% inhibitory concentration) for zidovudine > 1 microM and at least threefold greater than the IC50 of virus isolated from the matched nonprogressor (p = 0.04). In 16 of these 17 pairs the virus isolated from the progressor had the SI phenotype, indicative of high cytopathogenicity, while the virus from the matched nonprogressor had a non-syncytium-inducing phenotype (p < 0.0001). Zidovudine therapy did not appear to select for the SI phenotype in this patient population. A statistically significant association between high-level zidovudine resistance and clinical disease progression was demonstrated. A statistically significant association between the SI phenotype and disease progression was demonstrated. The results suggest that disease progression while being treated with zidovudine therapy may be more closely associated with the SI phenotype than with zidovudine resistance.

Acquired Immunodeficiency Syndrome↗

PubMed2001

Transfection of eastern oyster ( Crassotrea virginica) embryos.

There is a need for research in disease resistance and microbial elimination in the eastern oyster Crassosostrea virginica. Gene transfer may lead to advances in this area, and a means of selecting transfected larvae would be useful. We transfected 3-hour-postfertilization embryos with the bacterial gene aminoglycoside phosphotransferase II (neo(r)), which confers resistance to neomycin and related antibiotics such as G418. The antibiotic G418 was examined as a potential selective agent. A neutral red assay was used to determine survival after 48 hours of exposure to various concentrations of G418 (0-4 mg/ml). We examined the effects of electroporation and chemically mediated transfection of 3-hour-postfertilization embryos on survival to straight-hinge larvae. DNA alone was found to have no effect on survival (P >.05). For electroporation we found that increased voltage and pulse duration decreased survival (P <.05). Chemically mediated transfection did not significantly affect survival (P =.5172). Transgenic larvae were identified after electroporation and chemically mediated transfection. These larvae were reared for 24 hours and exposed to G418 at 0.3 mg/ml for 48 hours. Significant differences in survival between transfected and nontransfected larvae were detected for electroporation (P =.0147) and chemically mediated transfection (P =.037). Gene transfer was also confirmed with polymerase chain reaction and observation of expression of green fluorescent protein. This study documents the first successful insertion and expression of foreign DNA in eastern oyster larvae.

Journal Article↗

PubMed2000

Antibiotic susceptibility of Escherichia coli dnaK and dnaJ mutants.

The role of two chaperone proteins, DnaK and the cooperating factor DnaJ, in Escherichia coli antibiotic susceptibility to three antibiotics (a beta-lactam, chloramphenicol, tetracycline) has been studied. It was found that null dnaJ and dnaKdnaJ mutants are impaired in the functions leading to antibiotic susceptibility. The secretion of beta-lactamase to the periplasmic space is diminished in both mutants, and the additive effect of the two mutations was observed. The activity of chloramphenicol acetyltransferase is also impaired in an additive manner in both mutant strains. Tetracycline uptake is changed only in the double deletion mutant. These defects were observed only during incubation at high temperature (42 degrees C). Efficient complementation of some of these defects by the wild-type alleles introduced on low-copy number plasmid was achieved. Minimal inhibitory concentrations and the titer of the wild-type strains, delta dnaJ and delta dnaKdnaJ mutants treated with ampicillin, chloramphenicol, and tetracycline were also determined. Higher susceptibility of both mutants to chloramphenicol and tetracycline, as compared to their wild-type parent, was observed only after 1 h preincubation of cultures at 42 degrees C. On the contrary, both mutants were less susceptible to ampicillin than their parent strain.

Ampicillin↗

PubMed1982

[What therapeutic possibilities exist in acute antibiotic-resistant and chronic infections].

Since the rate of mortality in severe sepsis could not be changed in the last years even not by sophisticated antibiotics, and since the mutants of bacteria resistant to antibiotics are permanently increasing, other possibilities must be taken into considerations to prevent or to treat infections. The improvement of the patient's own immune resistance by active or passive immunizations seems to be a cooperative or alternative way to overcome severe and chronic infections. In animal experiments the efficacy of gammaglobulin treatment of a severe infection was tested. The positive result i.e. improvement of the mortality from 75% to 45%, stimulated to a controlled clinical study, in which severe infected patients were additionally treated with gammaglobulin and compared with those without gammaglobulin treatment. Patients with gammaglobulin treatment showed a much better outcome of their infection than control patients. The rate of infections as well as time of hospital stay and other clinical and laboratory parameters were markedly improved in gammaglobulin-treated patients. Also the principle of active immunization was tested in animal experiments. Guinea pigs were orally vaccinated with heat inactivated pathogenic bacteria. In a following challenge infection vaccinated animals survived in a high percentage whereas control animals died. This success in vaccinating animals did lead to a controlled clinical study with patients suffering from chronic bone infection. Patients were treated orally with heat inactivated bacteria over a time of 8 weeks. Not only laboratory data indicating an improved immune response were changed by the treatment but also the clinical findings. These results indicate clearly that infections can be treated not only by attacks against bacteria but also by the improvement of the patient's own bacterial resistance.

Animals↗

PubMed1999

Cell adhesion and the immune system: a case study using earthworms.

In the earthworm's immune system, cell adhesion, which occurs by putative receptors on leukocytes, is essential after recognition of self vs. non-self. Confrontation with foreign antigens is a normal event in the environment, replete with microbial pathogens that pose a threat to survival. To better understand what happens when an effector cell first recognizes a foreign target followed by its adhesion to it, isolated leukocytes, in sufficient quantities to be subjected to various analyses, have been extremely beneficial. In vitro approaches when accompanied by biochemical, immunological, and molecular technologies, have opened up new vistas concerning the immune response of earthworms and other invertebrates. The most recent discovery includes the preliminary identification of cell differentiation (CD) markers that play vital roles in recognitive and adhesive events. Certain leukocyte effectors show characteristics of natural killer (NK) cells that may act differently depending upon their source, whether autogeneic, allogeneic, xenogeneic, or expressed under normal or varying environmental conditions including exposure to xenobiotics. At the level of earthworm evolution, there is apparently a dissociation of phagocytosis from the process of killing by NK-like effectors. There are at least three future challenges. First, it is essential to determine the precise nature of the CD markers with respect to their molecular structure. Second, once their molecular and biochemical characteristics have been defined, the role of these markers in cellular and humoral mechanisms must be clarified in order to define effector cell products and resulting immune responses. Third, there is a need to differentiate between the several lytic factors that have been found in earthworms with respect to molecular structure, and biochemical and functional characterization.

Animals↗

PubMed1992

Use of a novel technique of cutaneous lavage for diagnosis of Lyme disease associated with erythema migrans.

OBJECTIVE: Determining the microbial cause of cellulitis is often difficult. In this study, a novel two-needle lavage technique was used to culture Borrelia burgdorferi from the skin of suspected erythema migrans lesions. DESIGN: The yield of lavage cultures for B burgdorferi was compared with that of a 2-mm skin biopsy sample. SETTING: A Lyme disease diagnostic center located in an area in which Lyme disease is epidemic. PATIENTS: Forty-five patients with suspected erythema migrans who had not been treated with antimicrobial agents. INTERVENTION: Cutaneous lavage of the advancing edge of a suspected primary erythema migrans lesion was done for all 45 participants, 33 of whom also had a skin biopsy of the same lesion at an identical (14) or an adjacent (19) site. MAIN OUTCOME MEASURES: Growth of B burgdorferi in in vitro culture. RESULTS: Lavage fluid cultures grew B burgdorferi in 13 (29%) of the 45 cases (95% confidence interval [CI], 16% to 44%). Among the 33 cases in which both lavage and skin biopsy cultures were done, the yield of lavage culture was less than that of biopsy culture (P less than .09, 12/33 vs 20/33). If contaminated cultures are excluded, this difference is significant (P less than .05, 12/30 vs 20/27). CONCLUSION: Cutaneous lavage is a new diagnostic technique for recovery of B burgdorferi from erythema migrans lesions that has potential applicability to other types of cutaneous infections.

Biopsy↗

PubMed2005

Microbiological and physicochemical factors affecting Aspergillus section Flavi incidence in Cavendish banana (Musa cavendishii) chips production in Southern Philippines.

Microbiological and physicochemical factors affecting the incidence of Aspergillus section Flavi in dried Cavendish banana (Musa cavendishii) chips production in Southern Philippines were examined. The average counts of Aspergillus section Flavi (AFC) in fresh and dried Cavendish bananas from 10 production batches of the Philippine Agro-Industrial Development Cooperative in Davao del Norte, Southern Philippines were 1.2 x 10(2) and 1.6 x 10(2) cfu/g, respectively. Isolates from both samples were identified to be Aspergillus flavus based on spore type and conidial structure of isolates. An increasing trend in the AFC of Cavendish bananas was observed during dried banana chips processing. Variability in the AFC between production batches was attributed to differences in aerobic and fungal populations and physicochemical characteristics of the fruits, peel damage of the raw materials, concentration of AFC in the air and food-contact surfaces of the production area, and temperature and relative humidity (RH) conditions of the environment during production and storage. Physicochemical characteristics of Cavendish bananas from the receipt of raw materials up to the first day of drying were within the reported range of values allowing growth and toxin production by aflatoxigenic fungi. Air-borne AFC varied depending on the section of the production area examined. The close proximity of the waste disposal area from the production operation to the preparation, drying and storage areas suggests that cross-contamination, probably air-borne or insect-borne was a likely occurrence. The hands of workers were also identified as AFC sources. Results of this study highlight the need for the development of strategies to control aflatoxigenic fungi and aflatoxin contamination in Philippine dried Cavendish bananas.

Air Microbiology↗