Antifungal activity of extracts and purified saponins from the rhizomes of Chamaecostus cuspidatus against Candida and Trichophyton species.

AIM: To evaluate the antifungal activity of extracts of Chamaecostus cuspidatus against Candida and Trichophyton species. METHODS AND RESULTS: Crude ethanol extracts of leaves, stems and rhizomes were prepared and evaluated for antimicrobial activity. Only the rhizomes extract (RE) showed antifungal activity but had no inhibitory effect against bacteria (Staphylococcus aureus and Escherichia coli). The RE was then submitted to liquid-liquid partition with hexane (Hex), dichloromethane, chloroform, ethyl acetate and water. The Hex fraction (Hex Fr) from the RE was found to have the best antifungal effect. Three known saponins were isolated from the Hex Fr, of which two (dioscin and aferoside A) showed good antifungal activity. In addition, Hex Fr and the two bioactive compounds had no antibacterial effect, but exhibited fungicidal activity, caused significant changes in the morphology of the fungal cells and showed anti-Candida albicans biofilm activity. Finally, the bioactive plant products presented greater selectivity for fungal cells over normal human cells. CONCLUSIONS: The rhizomes of C. cuspidatus have bioactive saponins that function as effective antifungals against Candida and Trichophyton species, and have antibiofilm activity against C. albicans. SIGNIFICANCE AND IMPACT OF THE STUDY: Chamaecostus cuspidatus REs may have potential clinical application towards the management of superficial mycoses caused by Candida and Trichophyton species.

Ex-vivo evaluation of the three-column concept in canine thoracolumbar fractures / [Avaliação em ex-vivo do conceito dos três compartimentos em fraturas toracolombares da coluna]

Traumatic events such as a motor vehicle accident or falling from heights are very common in veterinary medicine and often lead to vertebral fracture-luxation with concomitant spinal cord injuries, mostly in the thoracolumbar spine. The purpose of this cadaveric biomechanical study was to determine the feasibility of the three-column concept in canine thoracolumbar segments with induced fractures. Eighteen Functional Spinal Units (FSU) of the thoracolumbar segments (T12-L2) were collected from 18 medium-sized adult dog cadavers and were subjected to flexion-extension and lateral bending tests so that range of motion (ROM) was recorded with a goniometer. Fractures were induced by compressive loads applied by a universal testing machine (EMIC®). After this, specimens were screened using computed tomography (CT) and the fractures were graded as affecting one, two or three columns, and divided into groups A, B, and C, respectively. Post-fracture range of motion (ROM) was compared with the previous results. Groups B and C (with fractures in two or three columns) had instability in the two axes evaluated (P<0.05). The outcomes of this study support the applicability of the three-column theory to thoracolumbar spines of dogs, as the FSUs that suffered fractures in two or more columns showed axial instability.(AU)

Eventos traumáticos, como acidentes automobilísticos ou quedas, são muito comuns na medicina veterinária e, frequentemente, levam a fraturas ou luxações vertebrais, associadas a lesões medulares concomitantes, mais frequentemente na coluna toracolombar. O propósito deste estudo biomecânico em cadáveres foi determinar a viabilidade da teoria dos três compartimentos em fraturas induzidas em segmentos toracolombares de cães. Dezoito unidades espinhais funcionais (UEF) de segmentos toracolombares (T12-L2) foram coletadas de 18 cadáveres de cães adultos de médio porte e submetidas a testes de flexão-extensão e curvamento lateral, de modo que a amplitude de movimento (ADM) foi registrada com um goniômetro. Fraturas foram induzidas por meio de cargas compressivas aplicadas por uma máquina universal de ensaios (EMIC®). As amostras foram submetidas à tomografia computadorizada (TC), e as fraturas foram classificadas como afetando um, dois ou três compartimentos e divididas nos grupos A, B e C, respectivamente. A ADM pós-fratura foi comparada com os resultados prévios. Os grupos B e C (com fraturas em dois ou três compartimentos) apresentaram instabilidade nos dois eixos avaliados (P<0,05). Os resultados deste estudo corroboram a aplicabilidade da teoria dos três compartimentos para segmentos de coluna toracolombar em cães, uma vez que as UEF que sofreram as fraturas em dois ou mais compartimentos apresentaram instabilidade axial.(AU)

Ex-vivo evaluation of the three-column concept in canine thoracolumbar fractures / [Avaliação em ex-vivo do conceito dos três compartimentos em fraturas toracolombares da coluna]

Traumatic events such as a motor vehicle accident or falling from heights are very common in veterinary medicine and often lead to vertebral fracture-luxation with concomitant spinal cord injuries, mostly in the thoracolumbar spine. The purpose of this cadaveric biomechanical study was to determine the feasibility of the three-column concept in canine thoracolumbar segments with induced fractures. Eighteen Functional Spinal Units (FSU) of the thoracolumbar segments (T12-L2) were collected from 18 medium-sized adult dog cadavers and were subjected to flexion-extension and lateral bending tests so that range of motion (ROM) was recorded with a goniometer. Fractures were induced by compressive loads applied by a universal testing machine (EMIC®). After this, specimens were screened using computed tomography (CT) and the fractures were graded as affecting one, two or three columns, and divided into groups A, B, and C, respectively. Post-fracture range of motion (ROM) was compared with the previous results. Groups B and C (with fractures in two or three columns) had instability in the two axes evaluated (P<0.05). The outcomes of this study support the applicability of the three-column theory to thoracolumbar spines of dogs, as the FSUs that suffered fractures in two or more columns showed axial instability.(AU)

Eventos traumáticos, como acidentes automobilísticos ou quedas, são muito comuns na medicina veterinária e, frequentemente, levam a fraturas ou luxações vertebrais, associadas a lesões medulares concomitantes, mais frequentemente na coluna toracolombar. O propósito deste estudo biomecânico em cadáveres foi determinar a viabilidade da teoria dos três compartimentos em fraturas induzidas em segmentos toracolombares de cães. Dezoito unidades espinhais funcionais (UEF) de segmentos toracolombares (T12-L2) foram coletadas de 18 cadáveres de cães adultos de médio porte e submetidas a testes de flexão-extensão e curvamento lateral, de modo que a amplitude de movimento (ADM) foi registrada com um goniômetro. Fraturas foram induzidas por meio de cargas compressivas aplicadas por uma máquina universal de ensaios (EMIC®). As amostras foram submetidas à tomografia computadorizada (TC), e as fraturas foram classificadas como afetando um, dois ou três compartimentos e divididas nos grupos A, B e C, respectivamente. A ADM pós-fratura foi comparada com os resultados prévios. Os grupos B e C (com fraturas em dois ou três compartimentos) apresentaram instabilidade nos dois eixos avaliados (P<0,05). Os resultados deste estudo corroboram a aplicabilidade da teoria dos três compartimentos para segmentos de coluna toracolombar em cães, uma vez que as UEF que sofreram as fraturas em dois ou mais compartimentos apresentaram instabilidade axial.(AU)

Successful use of autogenous bone graft for the treatment of a radius-ulna nonunion in an amputee dog / Uso de enxerto ósseo autógeno no tratamento de uma não união óssea em um radioulna de um canino amputado

Fracture nonunions represent important complications in orthopedic surgeries. Nonunion repairs or bone defects are surgically challenging. Our aim was to describe a nonunion case, which was repaired with rapid bone recovery. An 8-month-old male mixed breed dog that has been previously operated was presented to the Veterinary Medical Teaching Hospital of São Paulo State University, with a right radius-ulna nonunion and an amputated contralateral forelimb. A cancellous bone graft was collected from a partially amputated limb, in order to correct the nonunion, and used in association with a locking plate. After four weeks, the bone graft had been incorporated into the original bone. Clinical union with good weight bearing was achieved after eight weeks.(AU)

As não uniões ósseas representam uma importante complicação nas cirurgias ortopédicas. Não uniões ósseas ou defeitos ósseos são um desafio na cirurgia. O objetivo do presente trabalho foi descrever um caso de não união óssea, que foi reparada de forma rápida. Um cão SRD de oito meses, macho, que havia sido operado previamente, deu entrada no Hospital Veterinário da Unesp - Jaboticabal com uma não união óssea em radioulna e o membro contralateral amputado. Um enxerto ósseo autógeno foi coletado do coto do membro contralateral amputado e utilizado junto a uma placa óssea bloqueada para o tratamento. Após quatro semanas, o enxerto ósseo estava incorporado ao osso original. A união clínica e o retorno normal à função foram alcançados após oito semanas.(AU)

Successful use of autogenous bone graft for the treatment of a radius-ulna nonunion in an amputee dog / Uso de enxerto ósseo autógeno no tratamento de uma não união óssea em um radioulna de um canino amputado

Fracture nonunions represent important complications in orthopedic surgeries. Nonunion repairs or bone defects are surgically challenging. Our aim was to describe a nonunion case, which was repaired with rapid bone recovery. An 8-month-old male mixed breed dog that has been previously operated was presented to the Veterinary Medical Teaching Hospital of São Paulo State University, with a right radius-ulna nonunion and an amputated contralateral forelimb. A cancellous bone graft was collected from a partially amputated limb, in order to correct the nonunion, and used in association with a locking plate. After four weeks, the bone graft had been incorporated into the original bone. Clinical union with good weight bearing was achieved after eight weeks.

As não uniões ósseas representam uma importante complicação nas cirurgias ortopédicas. Não uniões ósseas ou defeitos ósseos são um desafio na cirurgia. O objetivo do presente trabalho foi descrever um caso de não união óssea, que foi reparada de forma rápida. Um cão SRD de oito meses, macho, que havia sido operado previamente, deu entrada no Hospital Veterinário da Unesp - Jaboticabal com uma não união óssea em radioulna e o membro contralateral amputado. Um enxerto ósseo autógeno foi coletado do coto do membro contralateral amputado e utilizado junto a uma placa óssea bloqueada para o tratamento. Após quatro semanas, o enxerto ósseo estava incorporado ao osso original. A união clínica e o retorno normal à função foram alcançados após oito semanas.

Partial lateral corpectomy associated with pediculectomy to treat extradural chondroma in a dog: technique description / Corpectomia lateral parcial associada à pediculectomia no tratamento de condroma extradural em um cão: descrição da técnica

This study aimed to describe in detail the surgical technique of partial lateral corpectomy (PLC) associated with pediculectomy to treat an in vitro extradural chondroma. A 12-year old female Cocker Spaniel was seen in the hospital with proprioceptive ataxia of hind limbs associated with extradural compression between the 1st and 2nd lumbar vertebrae. The owner opted for euthanasia since the patient's condition changed due to simultaneous illnesses that culminated in a poor prognosis. The patient's body was formally ceded to perform experimental spinal decompression. The technique was effective to completely remove the epidural mass with minimal manipulation of the spinal cord...

Objetivou-se descrever detalhadamente a técnica cirúrgica de corpectomia lateral parcial associada à pediculectomia no tratamento de um condroma extradural in vitro. Uma cadela de 12 anos de idade, raça Cocker Spaniel, foi atendida com ataxia proprioceptiva dos membros pélvicos associada à compressão extradural entre a primeira e a segunda vértebras lombares. Devido a alterações secundárias decorrentes de afecções concomitantes que culminavam em um prognóstico desfavorável, o proprietário optou pela eutanásia do paciente, e o cadáver foi formalmente cedido para realização de descompressão medular experimental. A técnica utilizada foi eficaz na remoção completa da massa extradural, com mínima manipulação da medula espinhal...

Partial lateral corpectomy associated with pediculectomy to treat extradural chondroma in a dog: technique description / Corpectomia lateral parcial associada à pediculectomia no tratamento de condroma extradural em um cão: descrição da técnica

This study aimed to describe in detail the surgical technique of partial lateral corpectomy (PLC) associated with pediculectomy to treat an in vitro extradural chondroma. A 12-year old female Cocker Spaniel was seen in the hospital with proprioceptive ataxia of hind limbs associated with extradural compression between the 1st and 2nd lumbar vertebrae. The owner opted for euthanasia since the patient's condition changed due to simultaneous illnesses that culminated in a poor prognosis. The patient's body was formally ceded to perform experimental spinal decompression. The technique was effective to completely remove the epidural mass with minimal manipulation of the spinal cord.(AU)

Objetivou-se descrever detalhadamente a técnica cirúrgica de corpectomia lateral parcial associada à pediculectomia no tratamento de um condroma extradural in vitro. Uma cadela de 12 anos de idade, raça Cocker Spaniel, foi atendida com ataxia proprioceptiva dos membros pélvicos associada à compressão extradural entre a primeira e a segunda vértebras lombares. Devido a alterações secundárias decorrentes de afecções concomitantes que culminavam em um prognóstico desfavorável, o proprietário optou pela eutanásia do paciente, e o cadáver foi formalmente cedido para realização de descompressão medular experimental. A técnica utilizada foi eficaz na remoção completa da massa extradural, com mínima manipulação da medula espinhal.(AU)

Total vertebrectomy for stabilisation of chronic spinal lumbar luxation in a paraplegic dog without nociception.

An adult male crossbred dog was referred with a history of a road traffic accident that took place 1 month earlier. Neurological examination revealed paraplegia with absent nociception in the pelvic limbs. On epaxial palpation, significant curvature of the anatomical axis of the spine between the third and fourth lumbar vertebrae was observed, with the presence of a bone end almost piercing the dog's skin. Survey radiographs of the lumbar spine revealed severe dislocation between L3 and L4 vertebrae. During surgery, the spinal cord was not visible between the dislocated segments. Because of difficulties in reducing the lumbar luxation during surgery, vertebrectomy and vertebral shortening were performed. After alignment between vertebrae L3 and L5, eight cortical orthopaedic screws and bone cement were used for fixation. After 30 days, the dog started to use a wheelchair and was considered by its owner to have a good quality of life with no evidence of pain. To the authors' knowledge, this is the first case of severe luxation treated by total vertebrectomy and spine shortening in a dog. This surgery can be considered as an option in the management of severe spine luxation when the spinal cord is physically transected.

Ação da hidroesteira na cicatrização óssea de cães submetidos à técnica de avanço da tuberosidade tibial modificado / Hydromat action on bone healing in dogs submitted to technical tibial tuberosity advancement modified

O objetivo desse estudo foi avaliar os efeitos da hidroterapia na cicatrização óssea de cães submetidos à técnica de Avanço da Tuberosidade Tibial modificada (TTAm), por meio de avaliações radiográficas. Foram utilizados 10 cães de porte médio ou grande (Canis familiaris- Linnaeus, 1758), machos ou fêmeas, com diagnóstico de Ruptura de Ligamento Cruzado Cranial (RLCCr). Após o diagnóstico, os animais foram submetidos ao TTAm. Os cães foram separados em dois grupos experimentais contendo cinco animais em cada grupo: Grupo C (controle): animais não tratados com fisioterapia (hidroesteira); Grupo T (tratado): animais tratados com fisioterapia (hidroesteira). Para a avaliação da cicatrização óssea foi realizada a análise radiográfica no pós-operatório imediato, com 30, 60 e 90 dias pós-operatório verificando posicionamento de implante, presença de reação periosteal, imagens sugestivas de rejeição, infecção, densidade óssea (DO) e índice de mineralização (IM). Pôde-se visibilizar o padrão radiográfico de cicatrização óssea em ambos os grupos. Ao avaliar o IM não foi verificada diferença significativa entre animais tratados ou não com fisioterapia (P>0,05). Os cães tratados com fisioterapia (hidroterapia) por 15 dias, comparados com animais não tratados, tiveram uso funcional do membro precocemente e a amplitude de movimento maior. Concluiu-se que a utilização da hidroterapia como método auxiliar no tratamento dos cães submetidos à TTA modificada não promove diferença significativa na cicatrização óssea em comparação aos cães que não foram tratados com fisioterapia

The aim of this study was to evaluate the of hydromat action in bone healing in dogs undergoing modified TTA (TTAm) by means of radiographic evaluation. We used 10 dogs of medium or large (Canis familiaris, Linnaeus, 1758), male or female, diagnosed with Cranial Cruciate Ligament Rupture (CCLR). After the diagnosis, the animals underwent to TTAm. The dogs were divided into two experimental groups with five animals in each group: Group C (control): animals not treated with physiotherapy (hydrotherapy) and Group T (treated): animals treated with physiotherapy (hydrotherapy). For the evaluation of bone healing was evaluated radiographically in the immediate postoperative period, with 30, 60 and 90 days post-operatively by checking the positioning of implant, presence of periosteal reaction, images suggestive of rejection, infection, bone density ( DO) and the rate of mineralization (IM). We could visualize the radiographic pattern of bone healing in both groups. In assessing the IM was not significant difference between treated or not with physiotherapy (P> 0.05). Dogs treated with physiotherapy (hydrotherapy) for 15 days, compared with untreated animals had functional use of the limb and early range of motion greater. It was concluded that the use of hydrotherapy as an auxiliary method in the treatment of dogs undergoing modified TTA does not promote significant difference in bone healing compared with dogs not treated with physiotherapy

HYDROMAT ACTION ON BONE HEALING OF DOGS SUBMITTED TO MODIFIED TIBIAL TUBEROSITY ADVANCEMENT TECHNIQUE / Ação da hidroesteira na cicatrização óssea de cães submetidos à técnica de avanço da tuberosidade tibial modificada

O objetivo desse estudo foi avaliar os efeitos da hidroterapia na cicatrização óssea de cães submetidos à técnica de Avanço da Tuberosidade Tibial modificada (TTAm), por meio de avaliações radiográficas. Foram utilizados 10 cães de porte médio ou grande (Canis familiaris- Linnaeus, 1758), machos ou fêmeas, com diagnóstico de Ruptura de Ligamento Cruzado Cranial (RLCCr). Após o diagnóstico, os animais foram submetidos ao TTAm. Os cães foram separados em dois grupos experimentais contendo cinco animais em cada grupo: Grupo C (controle): animais não tratados com fisioterapia (hidroesteira); Grupo T (tratado): animais tratados com fisioterapia (hidroesteira). Para a avaliação da cicatrização óssea foi realizada a análise radiográfica no pós-operatório imediato, com 30, 60 e 90 dias pós-operatório verificando posicionamento de implante, presença de reação periosteal, imagens sugestivas de rejeição, infecção, densidade óssea (DO) e índice de mineralizaçã

Ação da hidroesteira na cicatrização óssea de cães submetidos à técnica de avanço da tuberosidade tibial modificado / Hydromat action on bone healing in dogs submitted to technical tibial tuberosity advancement modified

O objetivo desse estudo foi avaliar os efeitos da hidroterapia na cicatrização óssea de cães submetidos à técnica de Avanço da Tuberosidade Tibial modificada (TTAm), por meio de avaliações radiográficas. Foram utilizados 10 cães de porte médio ou grande (Canis familiaris- Linnaeus, 1758), machos ou fêmeas, com diagnóstico de Ruptura de Ligamento Cruzado Cranial (RLCCr). Após o diagnóstico, os animais foram submetidos ao TTAm. Os cães foram separados em dois grupos experimentais contendo cinco animais em cada grupo: Grupo C (controle): animais não tratados com fisioterapia (hidroesteira); Grupo T (tratado): animais tratados com fisioterapia (hidroesteira). Para a avaliação da cicatrização óssea foi realizada a análise radiográfica no pós-operatório imediato, com 30, 60 e 90 dias pós-operatório verificando posicionamento de implante, presença de reação periosteal, imagens sugestivas de rejeição, infecção, densidade óssea (DO) e índice de mineralização (IM). Pôde-se visibilizar o padrão radiográfico de cicatrização óssea em ambos os grupos. Ao avaliar o IM não foi verificada diferença significativa entre animais tratados ou não com fisioterapia (P>0,05). Os cães tratados com fisioterapia (hidroterapia) por 15 dias, comparados com animais não tratados, tiveram uso funcional do membro precocemente e a amplitude de movimento maior. Concluiu-se que a utilização da hidroterapia como método auxiliar no tratamento dos cães submetidos à TTA modificada não promove diferença significativa na cicatrização óssea em comparação aos cães que não foram tratados com fisioterapia(AU)

The aim of this study was to evaluate the of hydromat action in bone healing in dogs undergoing modified TTA (TTAm) by means of radiographic evaluation. We used 10 dogs of medium or large (Canis familiaris, Linnaeus, 1758), male or female, diagnosed with Cranial Cruciate Ligament Rupture (CCLR). After the diagnosis, the animals underwent to TTAm. The dogs were divided into two experimental groups with five animals in each group: Group C (control): animals not treated with physiotherapy (hydrotherapy) and Group T (treated): animals treated with physiotherapy (hydrotherapy). For the evaluation of bone healing was evaluated radiographically in the immediate postoperative period, with 30, 60 and 90 days post-operatively by checking the positioning of implant, presence of periosteal reaction, images suggestive of rejection, infection, bone density ( DO) and the rate of mineralization (IM). We could visualize the radiographic pattern of bone healing in both groups. In assessing the IM was not significant difference between treated or not with physiotherapy (P> 0.05). Dogs treated with physiotherapy (hydrotherapy) for 15 days, compared with untreated animals had functional use of the limb and early range of motion greater. It was concluded that the use of hydrotherapy as an auxiliary method in the treatment of dogs undergoing modified TTA does not promote significant difference in bone healing compared with dogs not treated with physiotherapy(AU)

HYDROMAT ACTION ON BONE HEALING OF DOGS SUBMITTED TO MODIFIED TIBIAL TUBEROSITY ADVANCEMENT TECHNIQUE / Ação da hidroesteira na cicatrização óssea de cães submetidos à técnica de avanço da tuberosidade tibial modificada.

O objetivo desse estudo foi avaliar os efeitos da hidroterapia na cicatrização óssea de cães submetidos à técnica de Avanço da Tuberosidade Tibial modificada (TTAm), por meio de avaliações radiográficas. Foram utilizados 10 cães de porte médio ou grande (Canis familiaris- Linnaeus, 1758), machos ou fêmeas, com diagnóstico de Ruptura de Ligamento Cruzado Cranial (RLCCr). Após o diagnóstico, os animais foram submetidos ao TTAm. Os cães foram separados em dois grupos experimentais contendo cinco animais em cada grupo: Grupo C (controle): animais não tratados com fisioterapia (hidroesteira); Grupo T (tratado): animais tratados com fisioterapia (hidroesteira). Para a avaliação da cicatrização óssea foi realizada a análise radiográfica no pós-operatório imediato, com 30, 60 e 90 dias pós-operatório verificando posicionamento de implante, presença de reação periosteal, imagens sugestivas de rejeição, infecção, densidade óssea (DO) e índice de mineralizaçã

In vitro activity of hypnophilin from Lentinus strigosus: a potential prototype for Chagas disease and leishmaniasis chemotherapy.

Hypnophilin and panepoxydone, terpenoids isolated from Lentinus strigosus, have significant inhibitory activity on Trypanosoma cruzi trypanothione reductase (TR). Although they have similar TR inhibitory activity at 10 µg/mL (40.3 µM and 47.6 µM for hypnophilin and panepoxydone, respectively; ~100%), hypnophilin has a slightly greater inhibitory activity (~71%) on T. cruzi amastigote (AMA) growth in vitro as well as on in vitro phytohemagglutinin (PHA)-induced peripheral blood mononuclear (PBMC) proliferation (~70%) compared to panepoxydone (69% AMA inhibition and 91% PBMC inhibition). Hypnophilin and panepoxydone at 1.25 µg/mL had 67% inhibitory activity onLeishmania (Leishmania) amazonensis amastigote-like (AMA-like) growth in vitro. The panepoxydone activity was accompanied by a significant inhibitory effect on PHA-induced PBMC proliferation, suggesting a cytotoxic action. Moreover, incubation of human PBMC with panepoxydone reduced the percentage of CD16(+) and CD14(+) cells and down-regulated CD19(+), CD4(+) and CD8(+) cells, while hypnophilin did not alter any of the phenotypes analyzed. These data indicate that hypnophilin may be considered to be a prototype for the design of drugs for the chemotherapy of diseases caused by Trypanosomatidae.

In vitro activity of hypnophilin from Lentinus strigosus: a potential prototype for Chagas disease and leishmaniasis chemotherapy

Hypnophilin and panepoxydone, terpenoids isolated from Lentinus strigosus, have significant inhibitory activity onTrypanosoma cruzi trypanothione reductase (TR). Although they have similar TR inhibitory activity at 10 μg/mL (40.3 μM and 47.6 μM for hypnophilin and panepoxydone, respectively; ~100 percent), hypnophilin has a slightly greater inhibitory activity (~71 percent) on T. cruzi amastigote (AMA) growth in vitro as well as on in vitro phytohemagglutinin (PHA)-induced peripheral blood mononuclear (PBMC) proliferation (~70 percent) compared to panepoxydone (69 percent AMA inhibition and 91 percent PBMC inhibition). Hypnophilin and panepoxydone at 1.25 μg/mL had 67 percent inhibitory activity onLeishmania (Leishmania) amazonensis amastigote-like (AMA-like) growth in vitro. The panepoxydone activity was accompanied by a significant inhibitory effect on PHA-induced PBMC proliferation, suggesting a cytotoxic action. Moreover, incubation of human PBMC with panepoxydone reduced the percentage of CD16+ and CD14+ cells and down-regulated CD19+, CD4+ and CD8+ cells, while hypnophilin did not alter any of the phenotypes analyzed. These data indicate that hypnophilin may be considered to be a prototype for the design of drugs for the chemotherapy of diseases caused by Trypanosomatidae.

Identification and characterization of bioemulsifier-producing yeasts isolated from effluents of a dairy industry.

New bioemulsifier-producing yeasts were isolated from the biological wastewater treatment plant of a dairy industry. Of the 31 bioemulsifier-producing strains, 12 showed emulsifying activity after 2months of incubation, with E(24) values ranging from 7% to 78%. However, only Trichosporon loubieri CLV20, Geotrichum sp. CLOA40, and T. montevideense CLOA70 exhibited high emulsion-stabilizing capacity, with E(24) values of 78%, 67%, and 66%, respectively. These isolates were shown to induce a strong emulsion stabilizing activity rather than the reduction of the interfacial tension. These strains exhibited similar growth rates in the exponential growth phase, with a clear acceleration after 24h and stabilization of the activity after 144h. Emulsification and stability properties of the bioemulsifiers were compared to those of commercial surfactants after the addition of NaCl and exposure to temperature of 100 degrees C. The compounds produced by the isolates appeared to be lipid-polysaccharide complexes. Gas chromatograph analysis of the lipidic fraction of the bioemulsifiers from CLV20, CLOA40, and CLOA70 shows the prevalence of (9Z,12Z)-octadeca-9,12-dienoic acid, in concentrations of 42.8%, 25.9%, and 49.8%, respectively. The carbohydrate composition, as determined by GC-MS of their alditol acetate derivatives, showed a predominance of mannose, galactose, xylose and arabinose.

Antifungal activities of compounds isolated from Piper abutiloides Kunth.

Piperaceae is a family of tropical plants known to have antifungal, antibacterial, tumour-inhibitory, antiviral, antioxidant, molluscicidal and leishmanicidal activities. In this work, extracts and fractions from aerial parts of Piper abutiloides (Piperaceae), a traditional medicinal plant, were evaluated against the fungal species Candida albicans, C. parapsilosis, C. krusei, C. glabrata, C. tropicalis, Cryptococcus neoformans and Sporothrix schenckii. The results have shown that the antifungal activity of this plant can be concentrated in the hexanic fraction after partitioning its hydroalcoholic extract between hexane and 90% aqueous methanol. The chromatographic fractionation of the bioactive part was monitored with a bioautographic assay using C. glabrata, and allowed the isolation of three antifungal compounds: pseudodillapiol, eupomatenoid-6 and conocarpan. These compounds presented different potencies against the fungi tested, with the strongest effect being observed for eupomatenoid-6 against C. glabrata, which presented a minimal inhibitory concentration value of 0.3 microg spot(-1). Conocarpan showed antifungal activity without apparent cytotoxic effect on normal human lymphocytes, as assessed by the proliferation assay with human peripheral blood mononuclear cells stimulated with phytohaemaglutinin. This work reveals for the first time the occurrence of these compounds in P. abutiloides and justifies further studies to clarify their mechanisms of action.

Efeito do miriadenolídeo isolado de Alomia myriadenia (Asteraceae) sobre o tumor de Erlich ascítico no camundongo / Effect of myriadenolide isolated from Alomia myriadenia (Asteraceae) on Ehrlich tumor in its ascitic form

Estudou-se atividade antineoplásica de um produto natural isolado de Alomia myriadenia (miriadenolídeo) no modelo do tumor de Ehrlich em camundongos. Dezoito fêmeas de camundongo Swiss foram inoculadas com 2x10(7) células viáveis de tumor de Ehrlich via intraperitoneal (0,3ml) e posteriormente distribuídas aleatoriamente em três grupos que receberam: grupo I (controle) - 0,3ml de solução de Hanks; grupo II - 31µg/kg de miriadenolídeo; e grupo III - 139µg/kg de miriadenolídeo. No oitavo dia de experimento, foram realizados exames hematológicos e perfil protéico sérico eletroforético. Coletou-se todo o líquido ascítico para avaliação do volume, aparência, pH, contagem de células viáveis e inviáveis, realização de esfregaços para contagem de células claras e escuras, leucócitos e avaliação das regiões organizadoras de nucléolos argentafins (AgNORs). Foram realizados exames macro e microscópicos do baço, fígado e rins e aspirado o conteúdo da medula óssea dos fêmures direito e esquerdo de cada animal para avaliação da relação mielóide:eritróide. Não houve diferença significativa no volume, pH, contagem de células viáveis e inviáveis entre os três grupos estudados, observando-se valores de 17,6 x 10(4) células tumorais viáveis no grupo III, 27,7 x 10(4) no grupo II e 21,1 x 10(4) no grupo I. As AgNORs apresentaram-se pequenas, com distribuição difusa e incontáveis no grupo I, e em menor quantidade no grupo III. Os animais do grupo III apresentaram a menor concentração protéica total sérica (4,7g/dl) (P<0,05) quando comparados com os do grupo II (5,3g/dl) e do grupo I (5,1g/dl). Os valores de albumina foram semelhantes nos três grupos (2,6g/dl), e as globulinas totais foram maiores (P<0,05) no grupo II (2,71g/dl) quando comparadas com os valores médios do grupo III (2,11g/dl) e semelhantes ao grupo I (2,43g/dl). Não houve diferença entre alfa e beta globulinas entre os três grupos estudados, porém as gamaglobulinas foram maiores...

Antitumoral activity of a natural product of Alomia myriadenia (myriadenolide) in Ehrlich tumor in mice was studied. Eighteen Swiss female mice were intra-peritoneal inoculated 2x10(7) viable cells of Ehrlich Tumor (0.3ml) and randomly distributed in three groups receiving via intra-peritoneal on the 3rd and 5th day post-inoculation the following treatments: group I (control) - 0.3ml Hanks solution; group II: 31µg/kg myriadenolide; and group III: 139µg/kg myriadenolide. On the eighth day of the experiment blood profile and protein serum electrophoresis were performed. All ascitic liquid was collected to evaluate the volume and pH; to observe the aspect; to count viable and no viable cells, dark and clear cells, leukocytes and nucleolar organizer regions (NORs). Macro and microscopic exams were performed and bone marrow was aspirated from right and left femurs of each animal to evaluate myeloid:erythroid ratio. It was not observed difference in volume, pH, counts viable and no viable cells in the groups, although group III showed smaller number of viable tumoral cells (17.6 x 10(4)) when compared to the group II (27.7 x 10(4)) and group I (21.1 x 10(4)). The investigation of NORs to evaluate the proliferative capacity of tumoral cells after myriadenolide treatment showed that cells were smaller, uncountable and with diffuse distribution in group I. They were in lower quantity in group III. These results suggest that myriadenolide in dose 139µg/kg (group III) delay the tumoral growing and, probably, cell proliferation. The animals of group III showed lower value of total protein (4.7g/dl) (P<0.05) when compared to animals from group II (5.3g/dl) and group I (5.1g/dl). The values of albumin were similar in all groups (2.6g/dl) and total globulin was higher (P<0.05) in group II (2.71g/dl) when compared to mean values of group III (2.11g/dl) and similar to group I (2.43g/dl). The decrease of total protein in group III occurred...

Efeito do miriadenolídeo isolado de Alomia myriadenia (Asteraceae) sobre o tumor de Erlich ascítico no camundongo / Effect of myriadenolide isolated from Alomia myriadenia (Asteraceae) on Ehrlich tumor in its ascitic form

Estudou-se atividade antineoplásica de um produto natural isolado de Alomia myriadenia (miriadenolídeo) no modelo do tumor de Ehrlich em camundongos. Dezoito fêmeas de camundongo Swiss foram inoculadas com 2x107 células viáveis de tumor de Ehrlich via intraperitoneal (0,3ml) e posteriormente distribuídas aleatoriamente em três grupos que receberam: grupo I (controle) - 0,3ml de solução de Hanks; grupo II - 31µg/kg de miriadenolídeo; e grupo III - 139µg/kg de miriadenolídeo. No oitavo dia de experimento, foram realizados exames hematológicos e perfil protéico sérico eletroforético. Coletou-se todo o líquido ascítico para avaliação do volume, aparência, pH, contagem de células viáveis e inviáveis, realização de esfregaços para contagem de células claras e escuras, leucócitos e avaliação das regiões organizadoras de nucléolos argentafins (AgNORs). Foram realizados exames macro e microscópicos do baço, fígado e rins e aspirado o conteúdo da medula óssea dos fêmures direito e esquerdo de cada animal para avaliação da relação mielóide:eritróide. Não houve diferença significativa no volume, pH, contagem de células viáveis e inviáveis entre os três grupos estudados, observando-se valores de 17,6 x 104 células tumorais viáveis no grupo III, 27,7 x 104 no grupo II e 21,1 x 104 no grupo I. As AgNORs apresentaram-se pequenas, com distribuição difusa e incontáveis no grupo I, e em menor quantidade no grupo III. Os animais do grupo III apresentaram a menor concentração protéica total sérica (4,7g/dl) (P<0,05) quando comparados com os do grupo II (5,3g/dl) e do grupo I (5,1g/dl). Os valores de albumina foram semelhantes nos três grupos (2,6g/dl), e as globulinas totais foram maiores (P<0,05) no grupo II (2,71g/dl) quando comparadas com os valores médios do grupo III (2,11g/dl) e semelhantes ao grupo I (2,43g/dl).(AU)

Antitumoral activity of a natural product of Alomia myriadenia (myriadenolide) in Ehrlich tumor in mice was studied. Eighteen Swiss female mice were intra-peritoneal inoculated 2x107 viable cells of Ehrlich Tumor (0.3ml) and randomly distributed in three groups receiving via intra-peritoneal on the 3rd and 5th day post-inoculation the following treatments: group I (control) 0.3ml Hanks solution; group II: 31µg/kg myriadenolide; and group III: 139µg/kg myriadenolide. On the eighth day of the experiment blood profile and protein serum electrophoresis were performed. All ascitic liquid was collected to evaluate the volume and pH; to observe the aspect; to count viable and no viable cells, dark and clear cells, leukocytes and nucleolar organizer regions (NORs). Macro and microscopic exams were performed and bone marrow was aspirated from right and left femurs of each animal to evaluate myeloid:erythroid ratio. It was not observed difference in volume, pH, counts viable and no viable cells in the groups, although group III showed smaller number of viable tumoral cells (17.6 x 104) when compared to the group II (27.7 x 104) and group I (21.1 x 104). The investigation of NORs to evaluate the proliferative capacity of tumoral cells after myriadenolide treatment showed that cells were smaller, uncountable and with diffuse distribution in group I. They were in lower quantity in group III. These results suggest that myriadenolide in dose 139µg/kg (group III) delay the tumoral growing and, probably, cell proliferation. The animals of group III showed lower value of total protein (4.7g/dl) (P<0.05) when compared to animals from group II (5.3g/dl) and group I (5.1g/dl). The values of albumin were similar in all groups (2.6g/dl) and total globulin was higher (P<0.05) in group II (2.71g/dl) when compared to mean values of group III (2.11g/dl) and similar to group I (2.43g/dl). The decrease of total protein in group III occurred by globulin reduction.(AU)

Clinical and laboratory evaluation of compliance in acute lymphoblastic leukaemia.

AIM: To evaluate compliance in children with acute lymphoblastic leukaemia (ALL). METHODS: Compliance was assessed through specific interviews, annotations from medical charts, and erythrocytic determination of 6-mercaptopurine metabolites. RESULTS: A total of 39 patients who had concluded maintenance phase of chemotherapy were included in the study. Mothers were responsible for delivering 6-MP in 87% of cases. Thirty five interviewees said that medical prescription was well understood and that the main reason for non-compliance was forgetfulness. Non-compliance was detected through interviews (33.3% of the cases), reports from medical charts (30.7%), and drug determination (16.6%); 53.8% of children were found to be non-compliant. Non-compliance was significantly associated with chronic undernourishment. Although not statistically significant, there was a trend for the group of non-compliant children to be associated with low per capita family income. No significant associations of non-compliance with age at diagnosis, gender, parents' schooling level, number of family members, power consumption, and medians of absolute leucocyte or neutrophil blood counts were detected. A short follow up period precluded valid analysis on outcome. In the non-compliant group (n = 21), seven children relapsed, contrasting with three relapses in the compliant group (n = 18). CONCLUSIONS: Results suggest that non-compliance is one of the mechanisms which underlies the adverse influence of socioeconomic factors on the outcome of children with ALL. Additional studies are necessary to confirm this hypothesis. Comprehensive approaches to the problem of non-compliance are urgently needed.

An improved HPLC method for the quantitation of 6-mercaptopurine and its metabolites in red blood cells

A procedure is described for the rapid determination of the intra-erythrocyte concentration of 6-mercaptopurine (6-MP) and its metabolites, 6-thioguanine nucleotides (6-TGN) and 6-methylmercaptopurine (6-MMP). Erythrocytes (8 x 10(8) cells) in 350 æl Hanks solution containing 7.5 mg dithiothreitol were treated with 50 æl 70 percent perchloric acid. The precipitate was removed by centrifugation (13,000 g) and the supernatant hydrolyzed at 100§C for 45 min. After cooling, 100 æl was analyzed directly by HPLC using a Radialpack Resolve C18 column eluted with methanol-water (7.5:92.5, v/v) containing 100 mM triethylamine. 6-TG, 6-MP and the hydrolysis product of 6-MMP, 4-amino-5-(methylthio)carbonyl imidazole, were monitored at 342, 322 and 303 nm using a Shimadzu SPD-M10A diode array UV detector. The analytes eluted at 5.3, 6.0 and 10.2 min, respectively. The calibration curves were linear (rý > 0.998), and the analytical recoveries were 73.2 percent for 6-TG, 119.1 percent for 6-MP and 97.4 percent for 6-MMP. The intra- and inter-assay variations were highest for 6-MP (9.6 and 14.3 percent, respectively). The lowest detectable concentrations were 3, 3 and 25 pmol/8 x 10(8) erythrocytes for 6-TG, 6-MP and 6-MMP, respectively. The quantification limits (coefficients of variation <15 percent) were 8, 10 and 70 pmol/8 x 10(8) erythrocytes for 6-TG, 6-MP and 6-MMP, respectively. The method was applied to the analysis of 183 samples from 36 children under chemotherapy for acute lymphoblastic leukemia. The concentrations of the metabolites in the red cells of the patients ranged from 0 to 1934 pmol/8 x 10(8) erythrocytes for 6-TGN, and from 0 to 105.8 and 0 to 45.9 nmol/8 x 10(8) erythrocytes for 6-MP and 6-MMP, respectively. The procedure gave results that were in agreement with those obtained with other methods designed to detect cases of non-compliance with treatment, including patient interviews and medical evaluation, among others, demonstrating its applicability to monitoring the treatment of leukemic children.