Effect of ethyl acetate crude extract of Lasiodiplodia pseudotheobromae IBRL OS-64 against oral cavity bacteria with emphasis on Streptococcus mutans

Streptococcus mutans and Streptococcus agalactiae have been reported in human oral diseases including cariesand periodontitis. Thus, the present study was performed to investigate the effect of ethyl acetate crude extract ofLasiodiplodia pseudotheobromae IBRL OS-64, an endophytic fungus isolated from Ocimum sanctum leaf againstS. mutans and S. agalactiae. Disk diffusion and broth microdilution assays were used to determine the antimicrobialactivities with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values.Meanwhile, the microscopic analysis was used to study the structural degeneration of bacterial cells. The fungal crudeextracts demonstrated favorable antibacterial activity toward both test bacteria and produced an inhibition zone rangingfrom 16.0 to 21.2 mm. The MIC and MBC values of the fungal crude extract toward S. mutans and S. agalactiae weredetermined and the results showed that the MIC and MBC values were in the range of 125–500 µg/ml and 125–1,000µg/ml, respectively. The time-kill study suggested that the ethyl acetate crude extract possessed bactericidal effectwith concentration and time-dependent. Structural degeneration studies revealed the major abnormalities occurredto S. mutans cells after treated with the fungal crude extract where completed alterations of their morphology withthe formation of cavities and collapsed cells beyond repair occurred. Therefore, the current study suggested the ethylacetate crude extract of L. pseudotheobromae IBRL OS-64 could be an effective antibacterial agent to treat oral cavitybacteria.

Somatic embryogenesis in Carica papaya as affected by auxins and explants, and morphoanatomical-related aspects

ABSTRACT The aim of this study was to evaluate somatic embryogenesis in juvenile explants of the THB papaya cultivar. Apical shoots and cotyledonary leaves were inoculated in an induction medium composed of different concentrations of 2,4-D (6, 9, 12, 15 and 18 µM) or 4-CPA (19, 22, 25, 28 and 31 µM). The embryogenic calluses were transferred to a maturation medium for 30 days. Histological analysis were done during the induction and scanning electron microscopy after maturing. For both types of auxin, embryogenesis was achieved at higher frequencies with cotyledonary leaves incubated in induction medium than with apical shoots; except for callogenesis. The early-stage embryos (e.g., globular or heart-shape) predominated. Among the auxins, best results were observed in cotyledonary leaves induced with 4-CPA (25 µM). Histological analyses of the cotyledonary leaf-derived calluses confirmed that the somatic embryos (SEs) formed from parenchyma cells, predominantly differentiated via indirect and multicellular origin and infrequently via synchronized embryogenesis. The secondary embryogenesis was observed during induction and maturation phases in papaya THB cultivar. The combination of ABA (0.5 µM) and AC (15 g L-1) in maturation medium resulted in the highest somatic embryogenesis induction frequency (70 SEs callus-1) and the lowest percentage of early germination (4%).

Anatomical and histochemical analysis of Dysphania ambrosioides supported by light and electron microscopy

ABSTRACT Dysphania ambrosioides (L.) Mosyakin & Clemants (syn: Chenopodium ambrosoides L.), Amaranthaceae, popularly known as “mastruz”, is an herb widely used in Brazil as anthelmintic. To contribute to the knowledge about medicinal plants, a microscopic analysis was accomplished to describe the main anatomical characters of root, stem, petiole and leaf blade of D. ambrosioides and histochemical tests were performed on the leaf blade. Cross-sections were obtained, by hand, for microscopic analysis of root, stem, petiole and leaf blade; to the leaf blade were still made paradermal sections, scanning electron microscopy analysis, maceration and histochemical tests. The main characters useful in the identification of the plant were: anomalous secondary thickening in the root and stem; presence of idioblasts containing crystal sand in the root, stem, petiole and leaf blade; in these there are also idioblasts with druses; presence of non-glandular and glandular trichomes in the stem, petiole and leaf blade; stomata on the stem, petiole and leaf blade, identified in these as anomocytic and anisocytic; dorsiventral mesophyll and collateral vascular bundles. Maceration revealed that the vessel elements are helical type. Through the histochemical tests, it was evidenced the presence of lipophilic substances, essential oils, oleoresins, phenolic compounds, starch, lignin and calcium oxalate crystals. This work provides support to the quality control of the species.

Nanometrology and its perspectives in environmental research

OBJECTIVES: Rapid increase in engineered nanoparticles (ENPs) in many goods has raised significant concern about their environmental safety. Proper methodologies are therefore needed to conduct toxicity and exposure assessment of nanoparticles in the environment. This study reviews several analytical techniques for nanoparticles and summarizes their principles, advantages and disadvantages, reviews the state of the art, and offers the perspectives of nanometrology in relation to ENP studies. METHODS: Nanometrology is divided into five techniques with regard to the instrumental principle: microscopy, light scattering, spectroscopy, separation, and single particle inductively coupled plasma-mass spectrometry. RESULTS: Each analytical method has its own drawbacks, such as detection limit, ability to quantify or qualify ENPs, and matrix effects. More than two different analytical methods should be used to better characterize ENPs. CONCLUSIONS: In characterizing ENPs, the researchers should understand the nanometrology and its demerits, as well as its merits, to properly interpret their experimental results. Challenges lie in the nanometrology and pretreatment of ENPs from various matrices; in the extraction without dissolution or aggregation, and concentration of ENPs to satisfy the instrumental detection limit.

The identities and anti-herpes simplex virus activity of Clinacanthusnutans and Clinacanthus siamensis

Objective: To distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities. Methods: Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted withn-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay. Results: Cross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) μg/mL, (44.50±2.66) μg/mL, (64.93±7.00) μg/mL, respectively where as those of C. siamensis were (60.00±11.61) μg/mL, (55.69±4.41) μg/mL, (37.39±5.85) μg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanolC. nutans leaves extracts were (72.62±12.60) μg/mL, (65.19±21.45) μg/mL, (65.13±2.22) μg/mL, respectively where as those of C. siamensis were (46.52±4.08) μg/mL, (49.63±2.59) μg/mL, (72.64±6.52) μg/mL, respectively. Conclusions: The combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.

The identities and anti-herpes simplex virus activity of Clinacanthus nutans and Clinacanthus siamensis

<p><b>OBJECTIVE</b>To distinguish the difference among the Clinacanthus nutans (Burm. f.) Lindau (C. nutans) and Clinacanthus siamensis Bremek (C. siamensis) by assessing pharmacognosy characteristics, molecular aspect and also to evaluate their anti-herpes simplex virus (HSV) type 1 and type 2 activities.</p><p><b>METHODS</b>Macroscopic and microscopic evaluation were performed according to WHO Geneva guideline. Stomatal number, stomatal index and palisade ratio of leaves were evaluated. Genomic DNA was extracted by modified CTAB method and ITS region was amplified using PCR and then sequenced. Dry leaves were subsequently extracted with n-hexane, dichloromethane and methanol and antiviral activity was performed using plaque reduction assay and the cytotoxicity of the extracts on Vero cells was determined by MTT assay.</p><p><b>RESULTS</b>Cross section of midrib and stem showed similar major components. Leaf measurement index of stomatal number, stomatal index and palisade ratio of C. nutans were 168.32±29.49, 13.83±0.86 and 6.84±0.66, respectively, while C. siamensis were 161.60±18.04, 11.93±0.81 and 3.37±0.31, respectively. The PCR amplification of ITS region generated the PCR product approximately 700 bp in size. There were 34 polymorphisms within the ITS region which consisted of 11 Indels and 23 nucleotide substitutions. The IC50 values of C. nutans extracted with n-hexane, dichloromethane and methanol against HSV-1 were (32.05±3.63) µg/mL, (44.50±2.66) µg/mL, (64.93±7.00) µg/mL, respectively where as those of C. siamensis were (60.00±11.61) µg/mL, (55.69±4.41) µg/mL, (37.39±5.85) µg/mL, respectively. Anti HSV-2 activity of n-hexane, dichloromethane and methanol C. nutans leaves extracts were (72.62±12.60) µg/mL, (65.19±21.45) µg/mL, (65.13±2.22) µg/mL, respectively where as those of C. siamensis were (46.52±4.08) µg/mL, (49.63±2.59) µg/mL, (72.64±6.52) µg/mL, respectively.</p><p><b>CONCLUSIONS</b>The combination of macroscopic, microscopic and biomolecular method are able to authenticate these closely related plants and both of them have a potency to be an anti-HSV agent.</p>

Etnobotánica, micrografía analítica de hojas y tallos y fitoquimica de Cuphea aequipetala Cav. (Lythraceae): una contribución a la farmacopea herbolaria de los Estados Unidos Mexicanos (FHEUM) / Ethnobotany, analytical micrograph of leaves and stems and phytochemistry of Cuphea aequipetala Cav. (Lythraceae): a contribution to the herbal pharmacopoeia of the United Mexican States (FHEUM)

Cuphea aquipetala Cav. (Hierba del cancer) is used in Mexican traditional medicine. In the present work, its most frequent medicinal applications were determined through interviews, revision of herbarium specimens, and bibliography. The quoted applications were: to treat pounds and wounds. For these purposes, the drug (leaf and stem) is prepared and administered as infusion, or as poultice. In order to provide data for inclusion of this species in the Herbal Pharmacopoeia of the United Mexican States (FHEUM) we describe its leaf architecture and diagnostic anatomical characters of drug,. The anatomical study revealed that the three types of trichomes observed can aid to differentiate this species from other Cuphea species. Anatomical variation was observed mainly in the midrib of the leaf. Leaf architecture is first described for this species, and provides the basis for future comparisons. In addition, the acetone-water extract of the aerial parts previously reported as cytotoxic activity against DU-145 (human prostate carcinoma) and HCT-15 (human colon carcinoma), showed the presence of polyphenolic compounds of the type of hydrolysable and condensed tannins, mannitol was isolated and identified.

Cuphea aquipetala Cav., Lythraceae (Hierba del cáncer) es utilizada en la medicina tradicional mexicana. En el presente trabajo se determinó mediante entrevistas, consultas de herbario y revisión bibliográfica sus aplicaciones médicas más frecuentes, a saber: para tratar golpes y heridas, para lo cual la droga (hoja y tallo) es preparada en infusión, o administrada en emplastos. Con la finalidad de aportar datos que ayuden a la inclusión de esta especie en la Farmacopea Herbolaria de los Estados Unidos Mexicanos (FHEUM), se describen la arquitectura foliar y los caracteres anatómicos de la droga. El estudio anatómico reveló que los tres tipos de tricomas reconocidos en la especie estudiada apoyan a diferenciarla de otras especies de Cuphea sp. Se observó variación anatómica principalmente en la nervadura central de la hoja. La arquitectura foliar se describe por primera vez para la especie y sienta las bases para futuras comparaciones. Por otra parte, el extracto acetona-agua de las partes aéreas que previamente ha sido reportado con actividad citotóxica contra células DU-145 (carcinoma de próstata humano) y HCT-15 (carcinoma de colon humano) mostró la presencia de compuestos polifenólicos del tipo de los taninos hidrolizables, taninos condensados, y se aisló manitol.

Microscopic Observation of Decomposition-inhibition Effect in GSE (Grapefruit Seed Extract) in Rat Liver / 체질인류학회지

Regarding to preserve the cadaver, formaldehyde has been used as a major preservative. However, the usage of formaldehyde has been considered by its harmful effects such as the disturbing ordor, toxicities and limitations to use. Therefore we studied the effect of decomposition-inhibition which is a natural product, grapefruit seed extract (GSE). Concerning the preservative activity, we sacrificed 8 week old male SD rat and collected liver. Using liver tissues, we treated GSE as a time dependant manner under 37degrees, 80+/-5% humidity conditions. To confirm GSE effects, we applied light and electron microscopic analysis. In results, we observed GSE attenuated the morphological changes and putrefaction of liver tissues more than 3 days. Herein, we introduced the potential substitute of formaldehyde to preserve the cadaver as well as animal tissues.

Padronizaçäo de métodos e quantificaçäo de matérias estranhas e fungos filamentosos. II. Geléias de frutas / Standardization of method and quantification of extraneous materials and mycelial of molds. II. Fuit jams

As geleias de frutas, assim como outros produtos alimentícios de origem vegetal, podem apresentar, como contaminantes, matérias estranhas e hifas de fungos fragmentadas, quando é utilizada matéria-prima inadequada ou não são adotadas boas práticas de fabricação. O objetivo do presente trabalho foi padronizar procedimentos analíticos para quantificação de filamentos micelianos de fungos totais e de Geotrichum, e de matérias estranhas em geleias de frutas e avaliar tais contaminantes em 117 amostras colhidas em supermercados da cidade de São Paulo. Foram adotados os métodos da Association of Official Analytical Chemists International (AOAC). Como resultado da padronização, foi estabelecida a diluição 1+6 para as geleias de abacaxi, ameixa, figo e uva na contagem Howard de filamentos micelianos. Quanto aos resultados das análises, 56,4 por cento das amostras continham matérias estranhas (principalmente fragmentos de insetos); 73,0 por cento filamentos micelianos pela contagem Howard (máximo de 83 por cento de campos positivos) e 23,0 por cento estavam positivas para hifas de Geotrichum. As geleias de abacaxi e de ameixa apresentaram menor número de amostras com os contaminantes estudados. Considerando-se a legislação de alimentos em vigor, os produtos estudados apresentaram alto índice de contaminação por filamentos micelianos e matérias estranhas. (AU)

Fruit jams as well as other nourishing products of vegetal origin may present as contaminantsextraneous materials and mold mycelial, when inadequate raw material is used or good manufacturepractices are not adopted. The objective of the present study was to standardize analytical procedures forquantification of mold mycelials and Geotrichum mold, and extraneous materials in fruits jams and toevaluate such contaminants in 117 samples of the products obtained from supermarkets in the city of SãoPaulo, Brazil. The methods of the Association of Official Analytical Chemists International (AOAC) wereadopted. As result of the standardization, a 1+6 dilution was established for the pineapple, plum, fig andgrape jams for the Howard mold counting. According to the results, 56.4% of the samples presentedextraneous materials (mainly insects fragments); 73.0% contained mycelial fragments for the Howardmold counting and 23.0% were positive for mycelial fragments of Geotrichum mold. The plum and pineapplejams presented the lowest number of samples with the studied contaminants. Considering the current foodlegislation, the evaluated products presented high index of contamination for mycelial fragments andextraneous materials. (AU)

Padronizaçäo de método e quantificaçäo de matérias estranhas e filamentos micelianos. I. Doces de frutas em pasta / Standardization of method and quantification of the extraneous materials and mycelial of mods. I. Fruit pastes in sweets

Com objetivos de padronizar procedimentos analíticos e avaliar a contaminaçäo por matérias estranhas e filamentos micelianos de fungos em doces de frutas em pastas industrializados (bananada, goiabada e marmelada), foram colhidas 57 amostras dos doces citados em supermercados da cidade de Säo Paulo. Os métodos utilizados para determinaçäo dos parâmetros estudados mostraram-se adequados. Quanto aos resultados das análises, 54,4 por cento das amostras apresentaram matérias estranhas (principalmente fragmentos de insetos); 80,7 por cento continham filamentos micelianos pela contagem Howard e 31,6 por cento estavam positivas para hifas de Geotrichum. Considerando a legislaçäo de alimentos em vigor, 83,0 por cento das amostras de doces em pasta estavam positivas em pelo menos uma das três determinaçöes, podendo concluir que os produtos analisados apresentaram alto índice de contaminaçäo, tendo como principal fator a presença de filamentos micelianos

In order to standardize analytical procedures and to evaluate the contamination for extraneousmaterials and mycelial of molds in industrialized fruits pastes in sweets (banana sweet paste, guava sweetpaste and quince sweet paste), 57 samples of such sweet products were gathered in supermarkets fromSão Paulo city, Brazil. The methods used for determination of the cited parameters revealed adequate. Inrelation to the results of the analyses, 54.4% of the samples presented extraneous materials (mainly insectsfragments); 80,7% contained mycelial fragments for the Howard mold counting and 31.6% were positivefor mycelial fragments of Geotrichum mold. Considering the food legislation in force, 83.0% of the fruitpastes in sweets were positive in at least one of the three determination, we can conclude that the analyzedproducts presented high index of contamination, having as main factor the presence of mycelial fragments

Diagnostic value of dipstick urinalysis as a screening test for urinary tract infection

BACKGROUND: Urinary tract infection is one of the most common diseases in primary practice. Although quantitative urine culture is the standard for diagnosis, a rapid screening test is needed for effective management. But numerous studies in the past were controversial. We evaluated the utility of dipstick urine analysis as a screening test for urinary tract infection. METHODS: A retrospective analysis of 311 patients who were admitted between May 1997, and April 1998 were chosen as subjects. Patients had a complete urine analysis, such as microscopic analysis, chemical analysis by dipstick and urine culture was performed as part of an evaluation for urinary tract infection or suspicious febrile illness. We compared dipstick analysis with microscopic examination and urine culture. The results of the dipstick urinalysis were compared with the results of the urine culture, sensitivity, specificity, positive predictive value and negative predictive value. RESULTS: A total of 311 cases were included in the study among which 122 cases were true positive in urine culture. For dipstick urine analysis 92 of 122 cases were leukocyte esterase positive and 39 of 122 were nitrite positive. On microscopic examination 44 of 122 cases were leukocyte positive and 40 of 122 were microorganism positive. CONCLUSION: The sensitivity of the dipstick urinalysis seems higher than the microscopic examination in urinary tract infection. We conclude that the dipstick urinalysis is a useful screening test for urinary tract infection for those above 2 years old.

Comparação entre métodos de extração de sujidades em farinha de rosca / Comparison between methods of light filth extraction in surphus-bread flour

Foram analisadas, através do exame microscópico, amostras de farinha de rosca, a fim de testar dois métodos de digestão e extração de sujidades: o método que utiliza a pancreatina e o que utiliza a solução de Tween-Versene. Concluiu-se que o método que utiliza a pancreatina é o mais sensível. A pesquisa de sujidades em 131 amostras do comércio, utilizando o método de digestão com pancreatina, levou à conclusão de que 69,4% das amostras estavam contaminadas com fragmentos de insetos e 19,80/0, com pêlos de roedores, demonstrando que as condições de produção desse tipo de alimento são bastante precárias (AU).

Verificação das condições higiênicas de biscoitos por microscopia / Verification of hyginec conditions of biscuits by microscopy

Foram analisadas pelo método da digestão com pancreatina, 109 amostras de biscoitos adquiridas no comércio da cidade de São Paulo, sendo 67 do tipo "Cream-Cracker" e 52 do tipo "Maizena". Chegou-se à conclusão que 100% dos biscoitos "Cream-Cracker" estavam em condições higiênicas insatisfatórias, contaminados com fragmentos de insetos, sendo 3,5% impróprios para o consumo por conterem pelos de roedores e que 96,15% dos biscoitos "Maizena" estavam em condições higiênicas insatisfatórias, sendo 5,77% impróprios para o consumo pelos mesmos motivos que os do tipo "Cream-Cracker" (AU).