Biases on the administered parenteral doses of an experimental drug during phase I clinical trials

Introduction: The pharmaceutical aspects of drug administration in clinical trials receive poor consideration compared with the important attention devoted to the analytical and mathematical aspects of biological sample exploitation. During PK calculations, many researchers merely use for dose the nominal amount declared, overlooking the noticeable biases that may result in the assessment of PK parameters. The aim of this work was to evaluate the biases related to doses injected of a biosimilar drug in 2 Phase I clinical trials. Patients (or Materials) and Methods: In trial A, 12 healthy volunteers received different doses of a biosimilar of interferon beta-1a by either subcutaneous (SC) or intravenous (IV) injection. The doses were prepared by partially emptying 0.5-mL syringes supplied by the manufacturer (drop count procedure). In trial B, 12 healthy volunteers received 3 different formulations of the drug by IV injection (biosimilar without albumin [HSA], biosimilar with HSA and original brand [Rebif®]) and 2 different formulations as multiple SC injections (biosimilar HSA-free and original brand). In both trials, the actual dose administered was calculated as: D = C·V - losses. The product titer C was assessed by ELISA. The volume administered IV was assessed by weighting. Losses were evaluated by in vitro experiments. Finally, the binding of 125I-interferon to HSA was evaluated by counting the free and HSA complexed molecule fractions separated by gel filtration. Results: Interferon was not significantly adsorbed onto the lines used for its IV administration. In trial A, the titer was very close to the one declared (96 ± 7%). In trial B, it differed significantly (156 ± 10% for biosimilar with/without HSA and 123 ± 5% for original formulation). In trial A, the dose actually administered showed a large variability. The real injected volume could be biased up to 75% compared with the theoretical volume (for the lower dose administered [ie, 0.03 mL]). This was mainly attributed to a partial re-aspiration of the drug solution before withdrawing the syringe needle. A strict procedure was therefore applied in trial B to avoid these inaccuracies. Finally, in trial B, 125I-Interferon beta-1a binding to HSA appeared time dependent and slow, reaching 50% after 16-hour incubation, which is close to steady state reported for the comparator Rebif®. Conclusion: These practical examples (especially biases on actual titer and volume injected) illustrate that actual dose assessment deserves attention to ensure accuracy for estimates of clearance and distribution volume in the scientific literature and for registration purposes, especially for bioequivalence studies.

Multisystem Langerhans' cell histiocytosis (Hand-Schüller-Christian disease) in an adult: a case report and review of the literature.

Langerhans' cell histiocytosis (LCH) is a rare and enigmatic clonal disorder that affects mainly children. It is characterized by single or multiple granulomatous mass lesions composed of cells with the Langerhans' cell phenotype. Clinical presentation and behavior are heterogeneous and can range from a solitary lytic bone lesion (i.e., eosinophilic granuloma) with a favorable course to a fatal disseminated leukaemia-like form, with a wide spectrum of intermediate clinical presentations between these two extremes. Although LCH typically involves the bone, lesions can be found in almost all organs. We are reporting the case of a multisystem LCH in a 47-year-old patient who presented with a panhypopituitarism and diabetes insipidus, and who, 5 years later, developed mandibular, mastoid and femoral lesions. The final diagnosis of LCH was made on mandibular biopsy.

A case of acute retinal pigment epithelitis: spectral domain optical coherence tomography time course and physiopathologic hypothesis.

PURPOSE: To report the time course of retinal morphologic changes in a patient with acute retinal pigment epithelitis (ARPE) using spectral domain optical coherence tomography (SD-OCT). METHODS: A 30-year old man was referred for blurred vision of his right eye after five days that appeared suddenly 15 days after recovery from a flu-like syndrome. SD-OCT was performed immediately, followed by fluorescein and infracyanine angiography at eight days and then at three weeks. RESULTS: At presentation, a bubble of sub-macular deposit was observed on the right macula with central golden micronodules in a honeycomb pattern. SD-OCT showed an "anterior dislocation" of all the retinal layers up to the inner/outer segment (IS/OS) line and irregular deposits at the OS level together with thickening of the retinal pigment epithelial (RPE) layer. As visual acuity increased, eight days later, the OCT showed reduction of the sub-retinal deposits and an abnormal hyperflectivity of the sub-retinal and RPE layers was observed. The patient showed a positive serology for picornavirus. DISCUSSION: The acute SD-OCT sections of this patient with ARPE were compared with histological sections of a 35 day old Royal College of Surgeons rat. Similar findings could be observed, with preservation of the IS/OS line and accumulation of debris at the OS level, suggesting that ARPE symptoms could result from a transient phagocytic dysfunction of the RPE at the fovea, inducing reversible accumulation of undigested OS. Picornaviruses comprising enterovirus and coxsachievirus described as being associated with acute chorioretinitis. In this case, it was responsible for ARPE. CONCLUSION: We hypothesize that ARPE syndrome results from a transient dysfunction of RPE, which can occur as a post viral reaction.

CARACTERIZAÇÃO PÓS-COLHEITA DE SEIS CULTIVARES DE CARAMBOLA (Averrhoa carambola L.)

Avaliaram-se as qualidades física e química dos frutos de seis cultivares de carambola (Averrhoa carambola L.), propagadas pela Estação Experimental de Citricultura de Bebedouro, São Paulo, objetivando a obtenção de informações sobre a qualidade dos frutos produzidos nesta região. Foram utilizados frutos das cultivares Tean-ma, Fwang Tung, Arkin, Malásia, Golden Star e Nota-10. Os frutos foram analisados quanto à massa fresca, comprimento, largura, volume, porcentagem de suco, polpa e semente, número de sementes, coloração (L, Cromaticidade e ângulo Hue). Na polpa, determinou-se o pH, assim como o teor de umidade, sólidos solúveis totais (SST), acidez total titulável (ATT), vitamina C, açúcares solúveis totais e redutores, cinzas, proteína, pectina total e solúvel, e minerais (Ca, P, K, Mg, S, Cu, Fe, Mn e Zn). Os resultados foram avaliados estatisticamente através de um delineamento inteiramente casualizado e revelaram que os melhores frutos são os originários das cultivares Arkin, Tean-ma e Nota-10, pois são os maiores, os mais pesados e apresentam os maiores teores de sólidos solúveis totais (SST) e açúcares solúveis (AST), assim como boa relação SST/ATT. Os frutos da 'Arkin' mostraram coloração mais amarela, enquanto os da 'Golden Star' foram considerados de coloração mais clara, ou com maior translucidez. A textura foi menor nos frutos da 'Arkin', e se relacionou com a maior solubilização de suas pectinas

Krikothyreotomie mit dem "Quicktrach"-Koniotomiebesteck [Cricothyreotomy using the Quicktrach coniotomy instrument set].

Percutaneous cricothyroidotomy may be a lifesaving procedure for airway obstruction, which cannot be relieved by endotracheal intubation and can be performed with specially designed instruments. A new device, the "Quicktrach", was evaluated by an anatomical preparation, flow and resistance measurements, and puncture of the cricothyroid membrane in 55 corpses. The size of the parts of the instrument (needle, plastic cannula, depth gauge) in relation to the size of the larynx is adequate, thus there is little likelihood of perforation of the posterior wall of the larynx. Resistance of the plastic cannula is sufficiently low to allow for adequate ventilation. The duration of time until the cannula is positioned properly in the trachea is significantly shorter, when an incision prior to the puncture is done (83 +/- 88 seconds without incision versus 35 +/- 41 seconds with incision; mean +/- SD). The "Quicktrach" is easy to apply even by inexperienced persons. The incidence of damage to the larynx (lesions including fractures of the thyroid, cricoid and 1. tracheal cartilage in 18%; soft tissue injury in 9%) is relatively high, however considering the live saving character of the procedure these numbers appear to be acceptable. Technical problems which occur with the use of the device are discussed and suggestions for improvement are made.

Percutaneous biopsy through the foramen ovale for parasellar lesions: surgical anatomy, method, and indications.

Knowledge of the pathological diagnosis before deciding the best strategy for treating parasellar lesions is of prime importance, due to the relative high morbidity and side-effects of open direct approaches to this region, known to be rich in important vasculo-nervous structures. When imaging is not evocative enough to ascertain an accurate pathological diagnosis, a percutaneous biopsy through the transjugal-transoval route (of Hartel) may be performed to guide the therapeutic decision.The chapter is based on the authors' experience in 50 patients who underwent the procedure over the ten past years. There was no mortality and only little (mostly transient) morbidity. Pathological diagnosis accuracy of the method revealed good, with a sensitivity of 0.83 and a specificity of 1.In the chapter the authors first recall the surgical anatomy background from personal laboratory dissections. They then describe the technical procedure, as well as the tissue harvesting method. Finally they define indications together with the decision-making process.Due to the constraint trajectory of the biopsy needle inserted through the Foramen Ovale, accessible lesions are only those located in the Meckel trigeminal Cave, the posterior sector of the cavernous sinus compartment, and the upper part of the petroclival region.The authors advise to perform this percutaneous biopsy method when imaging does not provide sufficient evidence of the pathological nature of the lesion, for therapeutic decision. Goal is to avoid unnecessary open surgery or radiosurgery, also inappropriate chemo-/radio-therapy.

The use of telomerase activity for the detection of prostatic cancer cells after prostatic massage.

PURPOSE: Prostate cancer is the most commonly diagnosed cancer in the United States. The diagnosis or followup of prostate cancer in men older than 50 years is based on digital rectal examination, measurement of the free-to-total prostatic specific antigen ratio and transrectal ultrasound assisted needle biopsy of the prostate. We developed and evaluated a noninvasive method for diagnosing prostate cancer based on the measurement of telomerase activity after prostatic massage in fresh voided urine or after urethral washing. MATERIALS AND METHODS: We obtained 36 specimens of cells after prostatic massage in the fresh voided urine of 16 patients who subsequently underwent radical prostatectomy and after urethral washing in 20 who underwent prostate needle biopsies. Ethylenediaminetetraacetic acid was immediately added to the collected urine or washing to a final concentration of 20 mM. After protein extraction by CHAPS buffer each specimen was tested for telomerase activity in a 2-step modified telomeric repeat amplification protocol assay. The 2 prostate cancer cell lines PC-3 and LNCaP with high telomerase activity were used as a positive control. RESULTS: Telomerase activity was detected in 14 of 24 samples with known prostate cancer (sensitivity 58%). In contrast, no telomerase activity was found in the 12 cases without histological evidence of prostate tumor (specificity 100%). Eight of 9 poorly differentiated cancers expressed telomerase activity (89%), while only 6 of 15 well and moderately differentiated cancers showed telomerase activity (40%). CONCLUSIONS: Our data illustrate that telomerase activity may be detected in voided urine or washing after prostatic massage in patients with prostate cancer. Sensitivity was higher for poorly differentiated tumors. This approach is not currently available for detecting prostate cancer in clinical practice. However, these results are promising and further studies are ongoing.

Resistência genética à podridão amarga em maçãs, determinada pela taxa de desenvolvimento da doença em frutos com e sem ferimentos

A maçã é um dos mais importantes produtos agrícolas de Santa Catarina e a segunda mais importante fruteira de clima temperado do Brasil. No entanto, a produção brasileira está alicerçada em cultivares importadas suscetíveis a diversas doenças. A podridão amarga causada pelo fungo Glomerella cingulata (Stoneman) Spaulding & Schrenk, (forma imperfeita Colletotrichum gloeosporioides (Penz.) Sacc.) é uma das mais importantes doenças de verão, podendo causar perdas muito elevadas. No presente trabalho, a inoculação artificial de C. gloesporioides em frutos com e sem ferimentos objetivou verificar a diferença de evolução da podridão amarga e identificar possíveis fontes de resistência nas seleções e novas cultivares de macieira desenvolvidas pela Epagri. Verificou-se ampla variação na reação de resistência entre as cultivares e seleções estudadas. O estabelecimento e o desenvolvimento da podridão amarga mostrou-se muito mais rápido através de ferimentos. As seleções M-6/00 e M-13/00 manifestaram resistência superior à das atuais cultivares Gala, Fuji e Golden Delicious. Essas seleções também apresentaram resistência superior à cv. Melrose, indicada como resistente em outros estudos.

Efectes de la poda mecànica del dosser foliar i les arrels sobre la productivitat i qualitat de fruits en pomera (Malus domestica Borkh.)

Els anys 2008 i 2009 es va dur a terme un assaig en una plantació de pomera cv. Golden Smoothee® per a avaluar l’efecte de la poda mecànica del dosser foliar i la poda mecànica d’arrels sobre el rendiment de l’arbre, el pes promig del fruit i la qualitat del fruit. Els resultats confirmen que la poda mecànica del dosser foliar no redueix el rendiment de fruits de qualitat comercial i tampoc afecta la qualitat dels fruits; la poda d’arrels quan es duu a terme durant l’hivern tampoc suposa cap reducció de rendiment ni de qualitat dels fruits de pomera.

Padrão de ocorrência de doenças em mamão armazenado sob atmosfera controlada

O objetivo deste trabalho foi avaliar a dinâmica de evolução de doenças pós-colheita do mamão armazenado sob atmosfera controlada, contendo 3,0% de O2 e 6,0% de CO2 ou 3,0% de O2 e 3,0% de CO2, em relação aos armazenados sob atmosfera ambiente, na simulação de condições de exportação refrigerada (10ºC) via marítima, e de comercialização no mercado europeu dos mamões 'Sunrise Solo' e 'Golden'. Constatou-se que a atmosfera controlada contendo 3% de O2 e 6% de CO2 promoveu maior perda de frutos, descartados devido à ocorrência das principais doenças pós-colheita: Antracnose, Podridão Peduncular, Mancha Chocolate e, principalmente, Pinta Preta, em ambas as cultivares. Acredita-se que a elevação da concentração de CO2 tenha provocado injúria fisiológica e que isto tenha favorecido a infecção dos tecidos e sua colonização.

Empirical models for predicting the production of wild mushrooms in Scots pine (Pinus sylvestris L.) forests in the Central Pyrenees

Mushroom picking has become a widespread autumn recreational activity in the Central Pyrenees and other regions of Spain. Predictive models that relate mushroom production or fungal species richness with forest stand and site characteristics are not available. This study used mushroom production data from 24 Scots pine plots over 3 years to develop a predictive model that could facilitate forest management decisions when comparing silvicultural options in terms of mushroom production. Mixed modelling was used to model the dependence of mushroom production on stand and site factors. The results showed that productions were greatest when stand basal area was approximately 20 m2 ha-1. Increasing elevation and northern aspect increased total mushroom production as well as the production of edible and marketed mushrooms. Increasing slope decreased productions. Marketed Lactarius spp., the most important group collected in the region, showed similar relationships. The annual variation in mushroom production correlated with autumn rainfall. Mushroom species richness was highest when the total production was highest.

Validação do método das notas para quantificação da incidência da mancha fisiológica do mamão através do uso de imagens digitais

Neste experimento, utilizou-se o software ImageJ versão 1.32j para quantificar a incidência da Mancha Fisiológica do Mamão (MFM) no intuito de aferir o método das notas, visto que o mesmo é um método subjetivo. O experimento consistiu de um delineamento inteiramente casualizado, em esquema fatorial 3x6, com três cultivares (Golden, Diva e Tainung 01) e seis notas (notas de 0 a 5, sendo 0 para a ausência de incidência da MFM e 5 para a ocorrência máxima de incidência da MFM), sendo os resultados interpretados estatisticamente por meio de análises de variância e de regressão. Foram amostrados 10 frutos para cada cultivar, dentro de cada nota, totalizando assim 180 frutos analisados, sendo os resultados avaliados através do programa Genes. Os resultados mostram que o método das notas pode ser utilizado com segurança, tendo o mesmo se mostrado bastante preciso para quantificar o nível de incidência da MFM.

Avaliação de diferentes tipos de atmosferas modificadas na vida útil de carambolas minimamente processadas

Este trabalho teve por objetivo a avaliação da vida útil potencial de carambolas cv. Golden Star, minimamente processadas, armazenadas em diferentes tipos de embalagens plásticas, para a concepção da atmosfera modificada. Os frutos colhidos fisiologicamente maturos apresentaram coloração verde-amarelada, sólidos solúveis (SS) médios de 6,8 ºBrix e massa média de 185 g. Antes da aplicação dos tratamentos, os frutos foram selecionados, higienizados em solução de hipoclorito de sódio (NaOCl) a 100 mg. L-1, resfriados por 12 horas a 15 ± 0,5 °C, seccionados transversalmente, sendo então novamente higienizados em solução de NaOCl a 10 mg.L-1, por 3 minutos. Em seguida, os pedaços em forma de estrelas foram acondicionados em bandejas rígidas de poliestireno, com capacidade para 250 g, e revestidas com os seguintes materiais: T1: filme plástico perfurado de polietileno de baixa densidade (PEBD), de 0,006 mm; T2: filme plástico poliolefínico com antiembassamento da Dupont® (AGF), de 0,015 mm; T3: filme plástico poliolefínico da Dupont® (HF), de 0,015 mm; T4: filme plástico de PEBD, de 60 µm; T5: filme plástico de PEBD, de 80 µm; T6: filme plástico de polipropileno (PP), de 22 µm, e T7: bandeja rígida de polietileno tereftalato (PET), com capacidade para 500 mL, com tampa do mesmo material. As embalagens foram então armazenadas em câmara frigorífica a 12 ± 0,5 °C e 90 ± 3% de U.R, por 18 dias. Observou-se que não houve diferença significativa quanto aos sólidos solúveis (SS) e acidez titulável (AT). O maior número de microrganismos e valor de pH foram observados nos frutos embalados nos filmes plásticos de PEBD de 0,006 mm, AGF e HF de 0,015 mm da Dupont®. Entretanto, os frutos acondicionados nas embalagens PET apresentaram o maior teor de ácido ascórbico. Da mesma forma, somente nas embalagens PET é que se conseguiu modificação atmosférica eficiente do ponto de vista da manutenção dos atributos de qualidade durante o AR. Assim, esse tratamento proporcionou adequado controle microbiológico e manutenção das características de qualidade por 18 dias para as carambolas minimamente processadas.

Aspectos morfológicos de folhas na diferenciação de variedades de carambola

Em certas espécies vegetais, a distinção entre variedades pode ser realizada com base em aspectos morfológicos das folhas, o que permite a diferenciação mesmo quando não apresentam flores e/ou frutos. O presente trabalho foi realizado objetivando a distinção entre variedades de caramboleira (Averrhoa carambola L.) através de aspectos foliares. Foram avaliadas quatro variedades de caramboleira: Arkin, B-10, Golden Star e Hart, com quatro repetições de 10 folhas cada. Pelos resultados obtidos no presente trabalho, conclui-se que há diferenças morfológicas foliares entre as variedades de caramboleira, permitindo a sua distinção, especialmente em condições de viveiro.