Peritonite piogranulomatosa por Nocardia concava em cão - relato de caso / Piogranulomatous peritonitis by Nocardia concava in dog - Case Report

Nocardiose é causada por bactérias do gênero Nocardia do subgrupo Actinomycetos, que são Gram-positivas aeróbicas, filamentosas e podem apresentar ramificações. O diagnóstico baseia-se na presença de lesão inflamatória, com o microrganismo morfologicamente compatível, associada ao isolamento e à identificação microbiológica e molecular. Este trabalho tem por objetivo relatar um caso de nocardiose em canino, que desenvolveu inflamação piogranulomatosa peritoneal seis meses após ovariossalpingo-histerectomia. O animal apresentava hipertermia, distensão abdominal, taquipneia, polidipsia, hiporexia, mucosas hipocoradas e fezes pastosas. Os achados laboratoriais evidenciaram anemia leve e leucocitose por neutrofilia com desvio à esquerda e hipoalbuminemia. Uma massa na região mesogástrica e efusão peritoneal foram evidenciadas por meio da ultrassonografia abdominal. O líquido foi classificado como exsudato piogranulomatoso, e o animal submetido à laparotomia exploratória para lavagem abdominal e remoção da massa. Após procedimentos terapêuticos, ocorreu piora clínica e óbito. Peritonite piogranulomatosa foi a principal alteração anatomopatológica a qual foi associada à Nocardia spp. Molecularmente, a espécie isolada se aproxima da N. concava, por meio da análise filogenética. Essa espécie já foi descrita como causa de infecção em humanos na Ásia, no entanto não há registros na literatura na espécie canina, sendo este o primeiro relato.(AU)

Nocardiosis is caused by an aerobic, gram-positive, ramificated and filamentous bacteria of the Nocardia genus, subgroup Actinomycetos. The diagnosis is based on the presence of the inflammatory lesions with the morphologically compatible microorganism associated with microbiological and molecular isolation and identification. The objective of this work is to report a case of nocardiosis in a canine that developed peritoneal pyogranulomatous inflammation six months after ovariosalpingohisterectomy. The animal had hyperthermia, abdominal distention, tachypnea, polydipsia, hyporexia, hypocorous mucosae and pasty feces. The laboratory findings revealed mild anemia and leukocytosis due to neutrophilia with left deviation and hypoalbuminemia. A mass in the mesogastric region and peritoneal effusion were evidenced by abdominal ultrasonography. The fluid was classified as pyogranulomatous exudate and the animal underwent exploratory laparotomy for abdominal lavage and mass removal. Despite the therapeutic procedures and clinical alterations the dog died. Piogranulomatous peritonitis was the main anatomopathological alteration which was associated with Nocardia spp. Molecularly, the isolated species approaches the N. concava species through phylogenetic analysis. This specie was described as a cause of infection in humans in Asia; however, there are no records in literature on the canine species, being this the first report.(AU)

Microbiota bacteriana de ovos de jacaré-do-pantanal incubados naturalmente / Bacterial microbiota of the Pantanal caiman eggs incubated in nature

A cadeia produtiva do Caiman yacare tem-se destacado no Mato Grosso com a exportação de 143.386 peles em 2015, cujo sistema de manejo (ranching) implica a incubação artificial dos ovos. Nesse processo, a contaminação bacteriana de ovos influencia a taxa de eclosão. O conhecimento da microbiota de ovos incubados naturalmente orienta o manejo sanitário adequado no incubatório. No presente estudo, são apresentadas informações sobre essa microbiota e sua correlação com a de outros crocodilianos, apontando-se as espécies com potencial patogênico. Amostras de 20 ninhos de C. yacare foram coletadas e semeadas em ágar sangue e ágar Mac Conckey. A colônia condizente com Salmonella sp. foi confirmada pela técnica de reação em cadeia de polimerase. Das 22 espécies bacterianas isoladas, 59% pertencem à família Enterobacteriaceae e 41% a outros táxons bacterianos. A semelhança dos achados com as bactérias isoladas na microbiota oral e/ou intestinal/cloacal de crocodilianos foi de 77,27%. As bactérias mais e menos frequentes foram, respectivamente, Bacillus cereus, Flavobacterium multivorum, Citrobacter freundii, Escherichia hermannii, Hafnia alvei, Morganella, morganni, Salmonella sp., Serratia marcescens e Shigella sonnei. Das bactérias isoladas, 86,36% têm potencial patogênico para crocodilianos. A origem materna e a ambiental da microbiota de ovos incubados naturalmente são, respectivamente, de 77,27% e 27,27%.(AU)

The Pantanal caiman productive chain has grown in Mato Grosso with the exportation of 143.383 leather pieces in 2015, whose management system (ranching) implies the artificial incubation of eggs. In this process, the bacterial contamination of the eggs influences the hatching rate. Knowledge of the naturally incubated microbiota of eggs guides the appropriate sanitary management in the incubator room. Here we present information about this microbiota and correlate it with that of other crocodilians, indicating the species with pathogenic potential. Samples were collected in 20 nests at Pantanal, and sown in blood and Mac Conckey agar. Salmonella sp. was confirmed through polymerase chain reaction technique. From the twenty-two different species of bacteria isolated, 59% are from the Enterobacteriaceae Family and 41% from other bacterial taxonomies. The similarity of findings to isolated bacteria in the crocodilians oral and/or intestinal/cloacal microbiota was of 77,27%. The most and least frequent bacteria were, respectively, Bacillus cereus, Flavobacterium multivorum and Citrobacter freundii and Escherichia hermannii, Hafnia alvei, Morganella, morganni, Salmonella sp., Serratia marcescens and Shigella sonnei. Among the isolated bacteria, 86,36% are pathogenic for crocodilians. The maternal and environmental origin of the microbiota of eggs naturally incubated is, respectively, of 77,27% and 27,27%.(AU)

Cladosporium cladosporioides isolado de gato com carcinoma de células escamosas / Cladosporium cladosporioides isolated from a cat with squamous cell carcinoma

Este relato descreve o caso do Cladosporium cladosporioides isolado de uma lesão periocular de um felino atendido no Hospital Veterinário da Universidade Federal de Mato Grosso, em Cuiabá. A principal queixa do proprietário era uma lesão periocular, com piora no decorrer do tempo, havia aproximadamente quatro meses. Foi descrita a tentativa de tratamento da lesão com anti-inflamatórios e antibióticos, sem sucesso. Na anamnese foi relatado que o animal tinha acesso à rua e a hábitos de caça e que não havia outros animais da casa com lesão semelhante. O animal foi submetido à biópsia e citologia para um diagnóstico mais preciso do caso. Um fragmento foi encaminhado para o Laboratório de Patologia Veterinária e outro para o de Microbiologia Veterinária. Nas análises histopatológicas, houve compatibilidade com carcinoma de células escamosas e, nas lâminas de citologia, foi evidenciado um processo inflamatório agudo. Nas características macroscópicas e microscópicas da colônia, houve compatibilidade com Cladosporium sp. Posteriormente, o DNA foi extraído e sequenciado, confirmando a espécie Cladosporium cladosporioides. O objetivo deste relato foi descrever o isolamento dessa espécie em um felino com carcinoma de células escamosas.(AU)

This report describes a case of Cladosporium cladosporioides isolated from a cat with a periocular lesion at the Veterinary Hospital, Cuiaba- Brazil. Owner described his animal as having a periocular lesion treated unsuccessfully with anti-inflamatories and antibiotics. During anamnesis, it was reported that the animal has access to the street, hunting habits and none of the other animals of the house had a similar injury. The animal underwent biopsy and cytology for more accurate diagnosis of the case. A fragment was referred to the Veterinary Pathology Laboratory and another for Veterinary Microbiology. In the histopathological analysis of biopsy, it was compatible with squamous cell carcinoma and the cytology slides showed an acute inflammatory process. Microbiogical analysis isolated fungus with Cladosporium sp. Subsequently, DNA was extracted and sequenced confirming Cladosporium cladosporioides species. This paper reports the isolation of this species in a feline with squamous cell carcinoma.(AU)

Novel macrolide-resistance genes, mef(C) and mph(G), carried by plasmids from Vibrio and Photobacterium isolated from sediment and seawater of a coastal aquaculture site.

UNLABELLED: The aim of this study was to determine whether mef(C) and mph(G), originally found on the transferable multi-drug plasmid pAQU1 from Photobacterium damselae subsp. damselae isolated from seawater of a fish farm, are responsible for conferring macrolide resistance. Since these genes are localized head-to-tail on pAQU1 and only four nucleotides exist between them, the single- and combination-effect of these genes was examined. When mph(G) alone was introduced to Escherichia coli, the minimum inhibitory concentrations (MICs) against erythromycin, clarithromycin and azithromycin increased, whereas introduction of mef(C) alone did not influence macrolide susceptibility. Introduction of both mef(C) and mph(G) dramatically increased the MICs to the same three macrolides, i.e. >512 µg ml(-1) , >512 µg ml(-1) and 128 µg ml(-1) respectively. These results suggest that the macrolide phosphotransferase encoded by mph(G) is essential for macrolide resistance, while the efflux pump encoded by mef(C) is required for high-level macrolide resistance. The tandem-pair arrangements of the mef(C) and mph(G) genes were conserved on plasmids ranging in size from 240 to 350 kb of the 22 erythromycin-resistant strains belonging to Vibrio and Photobacterium obtained from the fish farm. Sixteen of 22 plasmids ranged in size from 300 to 350 kb. This is the first report of novel macrolide resistance genes originating from a marine bacterium. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, mef(C) and mph(G) were found to be novel macrolide-resistance genes, and this is the first report of macrolide-resistance genes originating from a marine bacterium. These genes may be responsible for previously reported cases of the emergence of erythromycin-resistant bacteria in aquaculture sites by an unknown mechanism. The introduction of the tandem arrangement of the mef(C) and mph(G) genes in Escherichia coli increased the MICs to erythromycin, clarithromycin and azithromycin, suggesting a novel mechanism conferring high-level macrolide resistance via combined expression of the efflux pump and macrolide phosphotransferase.

Reappraisal of the taxonomy of Streptococcus suis serotypes 20, 22 and 26: Streptococcus parasuis sp. nov.

In order to clarify the taxonomic position of serotypes 20, 22 and 26 of Streptococcus suis, biochemical and molecular genetic studies were performed on isolates (SUT-7, SUT-286(T), SUT-319, SUT-328 and SUT-380) reacted with specific antisera of serotypes 20, 22 or 26 from the saliva of healthy pigs as well as reference strains of serotypes 20, 22 and 26. Comparative recN gene sequencing showed high genetic relatedness among our isolates, but marked differences from the type strain S. suis NCTC 10234(T), i.e. 74.8-75.7 % sequence similarity. The genomic relatedness between the isolates and other strains of species of the genus Streptococcus, including S. suis, was calculated using the average nucleotide identity values of whole genome sequences, which indicated that serotypes 20, 22 and 26 should be removed taxonomically from S. suis and treated as a novel genomic species. Comparative sequence analysis revealed 99.0-100 % sequence similarities for the 16S rRNA genes between the reference strains of serotypes 20, 22 and 26, and our isolates. Isolate STU-286(T) had relatively high 16S rRNA gene sequence similarity with S. suis NCTC 10234(T) (98.8 %). SUT-286(T) could be distinguished from S. suis and other closely related species of the genus Streptococcus using biochemical tests. Due to its phylogenetic and phenotypic similarities to S. suis we propose naming the novel species Streptococcus parasuis sp. nov., with SUT-286(T) ( = JCM 30273(T) = DSM 29126(T)) as the type strain.

Analysis of the factors affecting the formation of the microbiome associated with chronic osteomyelitis of the jaw.

Chronic osteomyelitis of the jaw (COMJ) is one of the most intractable diseases among head and neck infections. Antimicrobial agents are routinely administered for COMJ without sufficient bacterial information, resulting in frequent treatment failures. To improve our knowledge of the bacterial aetiology of COMJ and to assist in the development of effective treatments, we performed a comprehensive analysis of the microbiome. Sixteen patients with four clinical types of COMJ (four with suppurative osteomyelitis, three with osteoradionecrosis of the jaw, four with primary chronic osteomyelitis, and five with bisphosphonate-related osteonecrosis of the jaw) were enrolled in this study. Bone samples were subjected to bacterial community comparisons by 16S rRNA gene pyrosequencing. As a result, we clarified that COMJ was caused by a far greater range of bacterial species (12 phyla and 163 genera) than previously reported. Moreover, the bacterial structures in COMJ changed dramatically with disease stage and the condition of the affected bone. Multiple correlation analyses revealed that sequestration and bone exposure could affect the community structure. On the basis of these factors, we reclassified COMJ into three clinical stages: I, inflamed or sclerotic bone without exposure; II, sequestrum without exposure; and III, exposed sequestrum. In stage II, the bacterial diversity was significantly lower, and the anaerobe genera Fusobacterium, Tannerella (formerly Bacteroides) and Porphyromonas were more abundant, than observed during other stages. Because these bacteria habitually reside in any clinical stage, they were considered to constitute the core microbiome of COMJ. Targeting these bacteria should lead to the development of effective preventive measures and cures.

Treating ulcerative colitis by Adacolumn therapeutic leucocytapheresis: clinical efficacy and safety based on surveillance of 656 patients in 53 centres in Japan.

BACKGROUND/AIM: The Adacolumn selectively depletes granulocytes and monocytes/macrophages, which are thought to be part of the immunopathogenesis of ulcerative colitis. This work aims at evaluating the safety and clinical efficacy of the Adacolumn in patients with ulcerative colitis in large population-based data sets. METHODS: The Adacolumn post marketing surveillance in Japan was undertaken on 697 patients in 53 medical institutions over 7 years from 29 October 1999 to 28 October 2006. Clinical efficacy and safety data were provided by patients' physicians in the participating institutes. RESULTS: Safety was evaluated in all the 697 patients and efficacy in 656 patients. At entry, 92% of the patients were on salicylates, 74% on prednisolone and only 9% on immunomodulators. Approximately 40% of patients had severe ulcerative colitis and over 70% had ulcerative colitis that was refractory to conventional medications. There was no serious adverse events; mild to moderate adverse events were seen in 7.7% of the patients. The overall response (remission or significantly improved) was 77.3%; the remission rate based on clinical activity index was 71.1%, while 17.1% remained unchanged and 5.6% worsened. Patients were subgrouped into severe, moderate and mild ulcerative colitis based on clinical activity index (n=578), the response rates were 63.2%, 65.7% and 80.4%, respectively (P<0.001). Endoscopic assessment of efficacy showed very significant mucosal healing, Matts' endoscopic index improved from 3.2+/-0.04 to 2.1+/-0.7 (n=219, P<0.001); reduction in prednisolone dose (P<0.0001); leucocyte count (n=358, P<0.0001) and C-reactive protein (n=314, P<0.0001). Patients who received > or =6 Adacolumn sessions (n=319) did better than patients who received < or =5 sessions (n=188, P=0.004) and multivariate logistic regression analysis revealed that baseline granulocyte count was the strongest predictor of clinical response to Adacolumn (P=0.0191, odds ratio 1.151). CONCLUSION: This post marketing surveillance provides the largest ever efficacy and safety data on the Adacolumn therapeutic leucocytapheresis in patients with ulcerative colitis. As a non-pharmacologic treatment for patients with active ulcerative colitis most of whom were refractory to conventional drug therapy, the observed efficacy was very significant. Baseline granulocyte count was convincingly an independent predictor of clinical response.

Significance of additional high-dose cytarabine in combination with cyclophosphamide plus total body irradiation regimen for allogeneic stem cell transplantation.

The combination of cyclophosphamide (CY) and total body irradiation (TBI) has been used as a standard conditioning regimen for allogeneic transplantation. Several studies showed an advantage of adding high-dose cytarabine (HDCA) to this regimen. To clarify the significance of additional HDCA, we conducted a retrospective multicenter study and compared the clinical results of these two regimens. From June 1985 to March 2003, 219 patients with hematological malignancies underwent allogeneic transplantation after conditioning with CY+TBI 12Gy (n=73) or CA+CY+TBI 12Gy (n=146). Engraftment, overall survival, transplant-related mortality (TRM), relapse rate and incidence of graft-versus-host disease (GVHD) were compared according to risks and donors. Addition of HDCA had no impact on the relapse rate in all subgroups, and it was associated with lower TRM among standard-risk patients after related transplantation, and with higher TRM and worse survival among standard-risk patients after unrelated transplantation. The incidence of acute GVHD was not significantly different between the two regimens, and HDCA resulted in a higher incidence of chronic GVHD among standard-risk patients after related transplantation. In summary, addition of HDCA is not beneficial for high-risk patients, and is not recommended for standard-risk patients receiving unrelated transplantation.

Solid tumors after hematopoietic stem cell transplantation in Japan: incidence, risk factors and prognosis.

To evaluate the incidence, risk factors and prognosis for solid tumors after hematopoietic stem cell transplantation (HSCT) in Japan, 809 patients who had received HSCT between 1981 and 2000 were retrospectively analyzed. In all, 19 newly diagnosed secondary cancers were observed. The risk for cancer development was 2.8 times as high as that for expected cases. The cumulative incidence ratios at 5 and 10 years were 1.9 and 4.2%, respectively. The risk was significantly elevated for buccal cavity cancer (standard incidental ratio (SIR), 44.42: 95% confidence interval (CI) 17.86-91.51), esophageal cancer (SIR, 22.36: 95% CI 6.09-57.25), and cervical cancer (SIR, 8.58: 95% CI 1.04-31.01). Of 15 patients who developed solid cancers following allografting, 12 had chronic graft-versus-host disease (GVHD), and all 10 patients with squamous cell carcinoma of the buccal cavity or esophagus had chronic GVHD. On multivariate analysis, extensive chronic GVHD and age over 45 years at the time of transplantation were associated with a higher risk for solid cancers. In all, 17 patients received therapy for secondary cancers, nine of whom are still alive and the 5-year probability of survival from the diagnosis is 42.8%. Our data suggest that early detection of secondary cancers is very important in prolonging overall survival.

Cytomegalovirus antigenemia and outcome of patients treated with pre-emptive ganciclovir: retrospective analysis of 241 consecutive patients undergoing allogeneic hematopoietic stem cell transplantation.

CMV disease remains a major infectious complication after allogeneic hematopoietic stem cell transplantation (HSCT). To investigate the relationship between CMV antigenemia, treatment with ganciclovir (GCV), and outcome, we retrospectively analyzed 241 consecutive patients at risk for CMV infection who underwent allogeneic HSCT. Antigenemia-guided pre-emptive strategy with GCV was used for all patients. CMV antigenemia developed in 169 patients (70.1%), and CMV disease in 18 patients (7.5%). Multivariate analysis showed that acute GVHD (grades II-IV) was the only risk factor for developing antigenemia, and acute GVHD and advanced age for CMV disease. GCV use, as well as acute GVHD and advanced age, significantly increased the risk for bacterial and fungal infection after engraftment. Those who developed CMV antigenemia had a poorer outcome than those who did not (log-rank, P=0.0269), although the development of CMV disease worsened the outcome with only borderline significance (log-rank, P=0.0526). In conclusion, detection of antigenemia proved to be a poor prognostic factor for HSCT patients, which may be attributed to a combination of factors, including CMV disease itself, the effect of treatment, and a host status that allows for reactivation of CMV. Optimal pre-emptive strategy needs to be determined.

Immunologic rejection of the central graft after limbal allograft transplantation combined with penetrating keratoplasty.

PURPOSE: To study the incidence and prognosis of immunologic rejection of the central graft after limbal allograft transplantation (keratolimbal allograft transplantation [KLAT]) combined with penetrating keratoplasty (PKP). METHODS: Endothelial rejection in central penetrating graft after simultaneous KLAT and PKP using the same donor cornea was retrospectively studied. Incidence, reversibility, prognosis, and changes in limbal grafts were examined. RESULTS: Forty-five eyes underwent simultaneous PKP and KLAT. Endothelial rejection of the central graft was found in 16 eyes (35.6%). At last examination, 10 grafts (62.5%) restored clarity after immunosuppressive therapy. During rejection episodes, four eyes showed engorgement of vessels in limbal grafts, which existed before the episodes. Only one eye developed neovascularization with mild edema of the limbal grafts; however, no other limbal grafts showed abnormalities on biomicroscopy. No epithelial changes were noted, and 14 grafts (87.5%) were covered by corneal epithelium after the rejection. CONCLUSION: Approximately one third of eyes had endothelial rejection in the central graft after simultaneous KLAT and PKP. Abnormalities suggestive of rejection in the limbal grafts were seldom observed in these eyes, suggesting that immunologic response was different in central and limbal grafts.

New milbemycins from Streptomyces hygroscopicus subsp. aureolacrimosus: fermantation, isolation and strucutre elucidation.

Twelve new milbemycins have been isolated and characterized from some strains derived from Streptomyces hygroscopicus subsp. aureolacrimosus SANK 60286 and SANK 60526. The metabolites 1 approximately 4 and 9 approximately 11 were produced by strain RM28D-688 SANK 60797 as minor products. The metabolites 5 approximaetly 8 were obtained from a broth of strain 57-338 SANK 61796. Strain MK-1391 SANK 62896 was used for the production of metabolite 12. The new metabolites, eight alpha-class and four beta-class compounds, have new structural features. For example, milbemycins alpha26 and alpha27, have the 26-hydroxy moiety, and other derivatives (milbemycins alpha20 approximately 23) have different side chains at the C-26 position from those of milbemycins alpha11 and alpha14. In addition, 5-hydroxylmilbemycin beta7 (beta12), involved in the major biosynthetic pathway of 25-methyl and 25-ethyl milbemycins, was discovered.

[Retrospective study of acute myelogenous leukemia in 83 elderly patients: clinical and biological characteristics].

In order to characterize clinical and biological characteristics of elderly patients with acute myelogenous leukemia (AML), we retrospectively analysed 83 elderly patients aged 60 years or more and, as a control, 114 younger patients aged 15 to 59 years who were admitted to our hospital between August 1984 and January 1998. There was a significantly higher incidence of preceding myelodysplastic syndromes in the elderly patients. They also had a significantly higher incidence of unfavorable cytogenetic abnormalities (loss or partial deletion of chromosome 5 or 7) and a significantly lower incidence of favorable cytogenetic abnormalities, such as t(15:17), t(8:21), or inv(16). With regard to FAB subtypes in de novo AML, the incidence of M3 subtype was significantly lower in the elderly group. Myeloperoxidase positivity of AML cells in the elderly group was lower than that in the younger group. Laboratory data at presentation disclosed a lower peripheral leukemic cell count, a higher fibrinogen level, a lower serum protein level, and a higher serum creatinine level in the elderly group. They also had poorer performance status and more frequent concomitant diseases at presentation, including liver diseases, heart diseases, or documented infections. It was concluded that elderly AML patients 60 years or older had a higher incidence of poor prognostic factors compared to younger patients.

[Retrospective study of acute myelogenous leukemia in elderly patients: treatment and outcome of 83 consecutive patients].

We retrospectively analyzed treatments and outcomes for 83 acute myelogenous leukemia (AML) patients aged 60 years or more (median age 71) admitted to our hospital between August 1984 and January 1998. Complete remission was achieved in 36% of 78 patients who received anti-leukemic therapy, and median overall survival was 227 days. In addition to abnormal karyotypes involving chromosome 5 or 7, administration of less than 120 mg/m2/course of daunorubicin (DNR) during the initial treatment phase was an unfavorable prognostic factor for both CR and survival. Only 41% of all patients received 120 mg/m2/course of DNR or more, and had a significantly higher CR rate (56%) and longer survival, with a median of 389 days. It was suggested that intensive chemotherapy was effective for selected elderly AML patients who were relatively younger and had good performance status, although the number of such patients was limited in our study.

Bioconversion of milbemycin-related compounds: isolation and utilization of non-producer, strain RNBC-5-51.

A non-producing strain, the so-called strain RNBC-5-51 SANK 60198, was isolated during a screening program of strain improvement in the milbemycin production. Strain RNBC-5-51 indicated almost the same characteristics as those in the parent strain, that is, the abundant spore formation on agar media and the good growth in liquid media. But it does not produce any kind of milbemycins. In addition, strain RNBC-5-51 accumulated precursor-like compounds of milbemycin-polyketide, the production of which were inhibited by the addition of cerulenin. In the bioconversion experiments, strain RNBC-5-51 converted milbemycin beta6 and A4 to milbemycin alpha14, and milbemycin beta7 and A3 to milbemycin alpha11, respectively. This strain also converted milbemycin D and avermectin B1a, to 26-(3-methyl-2-butenoyloxy)milbemycin D and 26-(3-methyl-2-butenoyloxy)avermectin B1a, respectively. These results suggest that milbemycin alpha11 is biosynthesized through the same route as milbemycin alpha14, and the mutated step in strain RNBC-5-51 might be in the polyketide synthetic pathway of milbemycins. Strain RNBC-5-51 loses the ability for de novo synthesis of milbemycins, but it retains the ability to bioconvert the milbemycin skeleton. This strain might be useful for C-26 modification of milbemycin-related compounds.

Bioconversion of milbemycin-related compounds: biosynthetic pathway of milbemycins.

Streptomyces hygroscopicus subsp. aureolacrimosus SANK 60286 and SANK 60576 produce many kinds of milbemycins. Among them, milbemycin alpha11, alpha14, A3, and A4 have the most effective acaricidal activity. In this study, we investigated the terminal biosynthetic pathway to milbemycin alpha14 and A4 which accumulated as the final products in these strains. Using cerulenin, a specific inhibitor of fatty acid and polyketide biosynthesis, we conducted bioconversion experiments with cultures of several mutants, including milbemycin A4- and alpha14-producing strains. The bioconversions of milbemycin beta6 to milbemycin A4 and milbemycin A4 to milbemycin alpha14 could be identified. For the biosynthesis of milbemycin A4 from milbemycin beta6 in the milbemycin A4-high producing strain, there appeared to be two separate pathways exhibiting different sequences of furan ring formation and C-5 keto reduction steps.

Vibrio vulnificus septicemia in a patient with severe aplastic anemia.

A 53-year-old man with severe aplastic anemia developed sporadic Vibrio vulnificus septicemia 1 day after eating raw fish and shellfish. Although V. vulnificus infection is potentially fatal, he was saved by immediate and sensitive antibiotic administration. Patients with chronic hematologic disease are susceptible to infection by this organism and are prone to developing septicemia when they eat raw seafood. It is necessary for a patient with this infection to be given effective antibiotics as quickly as possible.

[Short-term intensive treatment for acute myelogenous leukemia (excluding M3 subtype) in adults].

Thirty-two adults (median age 48 years) with acute myelogenous leukemia (excluding M3) have been treated with short-term intensive therapy (M90 therapy). After induction therapy with daunorubicin, cytosine arabinoside (araC), 6-mercaptopurine, prednisolone, mitoxantrone (MIT) and etoposide (VP16), three regimens of post-induction chemotherapy were conducted as short an intercycle time as possible. The first regimen was with MIT and VP16, the second with behenoyl-araC and aclarubicin and the third with VP16, araC, vincristine and vinblastine. No further therapy was given. Complete remission was achieved in 24 (75%) of 32 patients and 24% of all patients were projected to remain free of disease at 5 years. The median duration of the entire therapy was 120 days with a range of 95 to 157 days. Post-induction regimens resulted in severe myelosuppression and their toxicity included treatment-related death in one patient. The treatment results of this short-term therapy were comparable to a former treatment protocol, M84 therapy with a median duration of the entire treatment therapy of 515 days. To confirm the advantages of such short-term therapy, prospective randomized comparisons with conventional post-induction therapy may be required.

Proliferative effects of several hematopoietic growth factors on acute myelogenous leukemia cells and correlation with treatment outcome.

The response of human acute myelogenous leukemia (AML) cells to four different hematopoietic growth factors (granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-1 beta (IL-1beta), interleukin-3 (IL-3), and stem cell factor (SCF)) and the relationship of the proliferative response of the AML cells to treatment outcome were studied. Proliferative responses were analyzed in 79 patients with de novo AML and 19 patients with AML arising from myelodysplastic syndrome (MDS). In de novo AML, a positive proliferative response (stimulation index >2) was seen in 65 to 75% of cases. AML cells arising from MDS had a much higher incidence of proliferative response to each growth factor (79 to 90%) and a much higher level of 3H-TdR incorporation. The relationship to treatment outcome was evaluated in 79 patients with de novo AML. The patients whose leukemic cells had a positive proliferative response to any growth factor, especially IL-3 and SCF, had a poorer outcome, ie a lower complete remission (CR) rate, shorter CR duration, and shorter survival. The outcome was particularly poor in patients whose leukemic cells had proliferative responses to all four or any of the growth factors, compared to patients whose leukemic cells had no response. This increased response may be a marker of poor prognosis in patients with AML.