Application of Comprehensive Genomic Profiling-Based Next-Generation Sequencing Assay to Improve Cancer Care in a Developing Country.

PURPOSE: Identifying actionable oncogenic mutations have changed the therapeutic landscape in different types of tumors. This study investigated the utility of comprehensive genomic profiling (CGP), a hybrid capture-based next-generation sequencing (NGS) assay, in clinical practice in a developing country. METHODS: In this retrospective cohort study, CGP was performed on clinical samples from patients with different solid tumors recruited between December 2016 and November 2020, using hybrid capture-based genomic profiling, at the individual treating physicians' request in the clinical care for therapy decisions. Kaplan-Meier survival curves were estimated to characterize the time-to-event variables. RESULTS: Patients median age was 61 years (range: 14-87 years), and 64.7% were female. The most common histological diagnosis was lung primary tumors, with 90 patients corresponding to 52.9% of the samples (95% CI 45.4-60.4%). Actionable mutations with FDA-approved medications for specific alterations correspondent to tumoral histology were identified in 58 cases (46.4%), whereas other alterations were detected in 47 different samples (37.6%). The median overall survival was 15.5 months (95% CI 11.7 months-NR). Patients who were subjected to genomic evaluation at diagnosis reached a median overall survival of 18.3 months (95% CI 14.9 months-NR) compared to 14.1 months (95% CI 11.1 months-NR) in patients who obtained genomic evaluation after tumor progression and during standard treatment (P = .7). CONCLUSION: CGP of different types of tumors identifies clinically relevant genomic alterations that have benefited from targeted therapy and improve cancer care in a developing country to guide personalized treatment to beneficial outcomes of cancer patients.

Evaluation of screening tests for antimicrobial residues in milk from individual cows treated with a combination of penicillins G and streptomycin / Valoración de pruebas de campo para detección rápida de residuos antimicrobianos en leche individual de vacas tratadas con una combinación de penicilinas y estreptomicina / Valoração de testes de campo para detecção rápida de resíduos antimicrobianos em leite individual de vacas tratadas com uma mistura de penicilinas e estreptomicina

Abstract Background: in Colombia, the law (Resolution 1382, 2013) prohibits the sale of milk that contains any antimicrobial drug residues, although no specific official screening tests and detection limits have been specified. At present, milk with positive results to both the Delvotest® and Snap® assays is simply rejected. To avoid contaminating bulk tanks with milk from individually treated cows, producers would benefit from having on-farm screening tests to conduct their own quality controls. In addition, on-site testing would allow farmers to check if the withdrawal times of commercially-available generic products are in accordance with labeled recommendations. Material and Methods: In this study, two commonly used rapid detection tests (Delvotest® and SNAP® specific for beta-lactams) were used on milk from 39 subclinical mastitic Holstein cows that were prescribed with daily intramuscular injections of a commercial suspension containing 8.000.000 IU of penicilin G (75% procaine penicilin G, 25% potasium penicillin) and 8 g of streptomycin sulfate, for a total of 4 days. Cows were individually milked and samples collected every 12 hours the day before, and for 3 days after the recommended withdrawal time of three days post-treatment. To inactivate the potential action of natural inhibitors of microbial growth that may be present in milk (ie., lysozyme and lactoferrin), the results of the Delvotest® were compared before and after milk samples were subjected to heat treatment (82°C for 5 minutes). Results: When the Delvotest® was used as per manufacturer's instructions (i.e., without heating), 7 of 39 cows were positive for one more day past the recommended withdrawal period. However, the results of the Snap® specific for beta-lactams and Delvotest® post-heating showed that only 2 of those 7 cows were positive, suggesting that 5 animals gave false positive results. For the 312 milk samples analyzed, a high degree of concordance was observed (Kappa coefficient=0.74±0.1) between the Snap® and Delvotest® post-heating. Conclusions: Considering that the streptomycin in this product is known to be eliminated faster than penicillin-G, the results suggest that the efficacy of the Delvotest® (after heat treatment) is similar to that of the Snap® beta-lactams for the detection of penicillin residues. However, when the Delvotest® was not preceded by heat treatment to inactivate potential natural inhibitors, it yielded a high number of false-positive results. The results also showed that in 95% (37/39) of the cows treated with this commercial product, the labeled instructions of a 3 day withdrawal period were adequate for compliance within the law.

Resumen En Colombia, la ley (Resolución 1382, 2013) prohíbe la venta de leche que contenga residuos de cualquier medicamento antimicrobiano, aunque no se especifican pruebas oficiales ni límites de detección que deban cumplir. Actualmente, la leche que emite resultados positivos a los kits del Delvotest® y Snap® simplemente es rechazada. Para evitar contaminar tanques de acopio con leche de vacas tratadas, los productores se beneficiarían de tener pruebas in situ que les permitan hacer sus propios controles. Además, ello permitiría comprobar si los tiempos de descarte de productos comerciales cumplen con las recomendaciones de los insertos. En este estudio se evaluaron dos pruebas de detección rápida (Delvotest® y SNAP® específico parar beta-lactámicos) en leche de 39 vacas con mastitis subclínica que fueron tratadas con inyecciones diarias intramusculares de una suspensión comercial de 8.000.000 UI de penicilina G (75% penicilina procaínica G, 25% penicilina potásica) y 8 g de sulfato de estreptomicina, durante 4 días. Las vacas se ordeñaron manualmente y recolectaron muestras de leche, cada 12 horas, por 1 día antes y 3 después del tiempo de retiro recomendable de 3 días post-tratamiento. Para inactivar la acción de inhibidores naturales del crecimiento bacteriano que pueden estar presentes en leche (ej, lisozima y lactoferrina), los resultados del Delvotest® se compararon antes y después de que las muestras de leche fuesen sometidas a calentamiento (82°C por 5 minutos). Cuando el Delvotest® se usó de acuerdo a las instrucciones de la compañía, es decir, sin calentamiento, 7 de 39 vacas dieron positivas por ≥ 1 días pasado el tiempo de retiro recomendado. Sin embargo, los resultados del Snap® y Delvotest® post-calentamiento mostraron que sólo 2 de las 7 vacas eran positivas, sugiriendo que 5 animales estaban dando falsos positivos. En las 312 muestras de leche analizadas se obtuvo un alto grado de concordancia (coeficiente Kappa = 0.74±0.1) entre el Snap® y el Delvotest® post-calentamiento. En conclusión, los resultados sugieren que la eficacia del Delvotest® (post-calentamiento) es similar a la del Snap® específico para beta-lactámicos en lo que respecta a detección de residuos de penicilina. Sin embargo, cuando el Delvotest® no iba precedido de calentamiento para inactivar inhibidores naturales, se produjeron demasiados falsos positivos. Los resultados también mostraron que en el 95% (37/39) de las vacas tratadas con este producto, la recomendación de descarte por 3 días cumplía con la legislación vigente de no contener residuos.

Resumo Na Colômbia, a lei (Resolução 1382 de 2013) proíbe a venda do leite que tenha resíduos de qualquer medicamento antimicrobiano, embora não se especifiquem testes oficiais nem limites de detecção que devam se cumprir. Atualmente, o leite que emite resultados positivos aos Kits de Delvotest® e Snap® simplesmente é rejeitado. Para evitar contaminar os tanques de armazenamento com leite de vacas tratadas, os produtores beneficiar-se-iam de ter os testes in situ que lhes permita fazer seus próprios controles. Além, isto permitiria comprovar se os tempos de retiro do leite ao utilizar produtos comerciais cumprem com as recomendações da vide bula. Neste estudo avaliaram-se dois testes de detecção rápida (Delvotest® e SNAP® especifico parar beta-lactâmicos) no leite de 39 vacas com mastite subclínica que foram tratadas com injeções diárias intramusculares de uma suspensão comercial de 8.000.000 UI de penicilina G (75% penicilina procaína G, 25% penicilina potássica) e 8g de sulfato de estreptomicina durante quatro dias. Extraiu-se o leite das vacas com ordenha manual e se fez uma amostragem de leite (cada 12 horas) um dia antes e três dias depois do tempo de retiro do leite, que tinha como recomendável, na vide bula, três dias após final do tratamento. Para inativar a ação de inibidores naturais do crescimento bacteriano, que possam estar presentes no leite (ex. lisozima e lactoferrina), os resultados do Delvotest® compararam-se antes e depois de que as amostras de leite fossem sometidas a aquecimento (82°C durante 5 minutos). Quando o Delvotest® usou-se de acordo com as instruções da companhia, quer dizer, sem aquecimento, sete das 39 vacas deram positivas por ≥ 1 dia passado o tempo de retiro recomendado. Embora, os resultados do Snap® e Delvotest® após aquecimento do leite mostraram que só dois das sete vacas eram positivas, sugerindo que cinco animais estavam apresentando falsos positivos. Nas 312 amostras de leite analisadas obteve-se um alto grau de concordância (coeficiente Kappa = 0.74±0.1) entre o Snap® e o Delvotest® após aquecer o leite. Em conclusão, os resultados sugerem que a eficácia do Delvotest® (após o aquecimento do leite) é similar á do Snap ou especifico para beta-lactâmicos no que respeita a detecção de resíduos de penicilina. Embora, quando o Delvotest® não ia precedido do aquecimento do leite para inativar inibidores naturais, produziram-se muitos falsos positivos. Os resultados também demonstraram que o 95% (37/39) das vacas tratadas com este produto, estavam de acordo com a recomendação de descarte por três dias e cumpria com a legislação vigente de não conter resíduos.

Minimal-change disease as a paraneoplastic syndrome in a patient with ovarian carcinoma.

Minimal-change disease (MCD) is an exceptional paraneoplastic presentation. We are describing the case of an ovarian paraneoplastic nephrotic syndrome. The kidney biopsy was consistent with MCD. Steroids and immunosuppressive therapy were given with no change in the nephrotic-range proteinuria. A complete resolution of the nephrotic syndrome was soon observed with improvement of her clinical condition after five cycles of chemotherapy with paclitaxel and carboplatin and tumor-debulking surgery. Ovarian carcinoma paraneoplastic nephrotic syndrome secondary to MCD is an extremely rare event, which is important to recognize since it is responsive to the standard chemotherapy.

Polymorphic drug metabolism in anaesthesia.

A substantial part of the interindividual variability in response to drugs and xenobiotics is related to genetically-determined impairment in drug metabolism. Several drug-metabolising enzymes are polymorphic in humans and often polymorphisms are strongly related to altered drug biodisposition and to the risk of developing adverse effects. Drugs used in general anaesthesia undergo polymorphic metabolism. Among these, halothane is metabolized by cytochrome P450 (CYP) 2E1 and, to a lesser extent, by CYP3A4 and CYP2A6. CYP2E1 also plays a key role in the metabolism of isoflurane, sevoflurane, enflurane and desflurane. CYP2B6, CYP3A4 and CYP2C9 play a relevant role in the metabolism of ketamine. The enzymes involved in the metabolism of thiopental and etomidate remains to be elucidated. Propofol is metabolized mainly by glucuronidation by uridine diphosphate-glucuronosyltransferases (UGTs) and by hydroxylation by CYP2B6 and CYP2C enzymes. The enzymes SULT1A1 and NQO1 participate in later steps in propofol metabolism. All the above-mentioned anaesthetic-metabolising enzymes are polymorphic in man. The present review analyzes the importance of enzymes in the metabolism of anaesthetics and common polymorphisms related to the biotransformation of general anaesthetics and it raises hypotheses on genetic and non-genetics factors related to altered response to anaesthetics that require further investigation. Based on functional relevance and allele frequencies, we identify the most promising targets for the clinical use of pharmacogenomic techniques in anaesthesia to prevent altered pharmacokinetics or adverse drug effects.

Evaluation of propofol as an anesthetic in swine tracheal transplant surgery / Evaluación del propofol como anestésico para cerdos sometidosa trasplante de tráquea

Because the swine have been used as an ideal animal model for different medical investigations, it has been useful to the advance in vital organs transplant field. The trachea transplant is a surgical procedure which requires special conditions in anesthetic depth and muscular relax, for a long period, and in addition, an excellent intra and post-operatory analgesic. The aim of this study was to use a combination of xylacine and ketamine, as premedication and evaluate propofol as a general anesthetic in trachea transplant donor or recipient pigs. All the methodology was under the approval of the Committee of Ethics for the Experimentation with Animals of the University of Antioquia. Ten donors and 10 recipients female Yorkshire pigs having a body weight of about 30 kg were used. Trachea extraction from a donor and its transplantation to a recipient in the same surgical procedure was performed. The average body weight (PP) was 30 ± 2.92 kg for both the groups, the average value were as follows: time of recumbency (TR) 8.25 ± 2:85 min; latency period (PL) 6.05 ± 1.73 min, (for both groups); surgical time (TQ) for donors and recipients was 80 ± 0.02, and 247 ± 0.02 min, respectively; heartbeat rate (FC) 90.34 ± 8.14 bpm, O2 saturation (SO2) 95.47 ± 1.79 %; exhaled PCO2 31.13 ± 1.89 mmHg; temperature (T) for both groups was 37.51 ± 0.74oC. The mean arterial pressure average (PAM) for both group was 65.47 ± 5.94 mmHg; the average time of esternal recumbecency (TRE) for donor female pigs was 16.50 ± 4.09 min, and the average time to stand up (TP) for swine recipients was 30.70 + 3.27 min. These results indicate that Propofol can be considered as a safe anesthetic for use in continuous perfusion. Since it has not an analgesic effect it is strongly recommended to combine it with opioids during anesthetic-surgical procedures; it can be also used with neuromuscular preanesthetics or inhaled anesthetics.

Los cerdos se han utilizado como modelo animal ideal para diversas investigaciones médicas; han sido útiles para el avance en el trasplante de órganos. El trasplante de tráquea es un procedimiento quirúrgico que requiere condiciones especiales en profundidad anestésica y relajación muscular por un período largo, y además, una analgesia intra y del postoperatoria excelente. Nuestra investigación utiliza una combinación de xylacine y ketamina, como premedicación y evaluar el propofol como anestésico general en cerdos donantes y receptores en quienes el trasplante de la tráquea sería hecho. Toda la metodología contó con la aprobación del Comité de Ética para la Experimentación con los Animales de la Universidad de Antioquia. Utilizamos 10 donantes y 10 cerdos raza Yorkshire hembras con un peso corporal de cerca de 30 kilogramos. Se realizó la extracción de la tráquea de un donante y el trasplante a un receptor en el mismo procedimiento quirúrgico. El peso corporal (PP) fue de 30 ± 2.92 kg para todo el grupo, el tiempo de recumbencia (TR) para ambos grupos fue de 8.25 ± 2.85 min, el período de latencia (PL) para ambos grupos fue de 6.05 ± 1.73 min, el promedio de tiempo quirúrgico (TQ) para los donantes fue de 80 min ± 0.02, el TQ de los receptores fue de  247m ± 0.02. La presión arterial media (PAM) para todo el grupo fue de 65.47 ± 5.94 mmHg, el promedio de frecuencia cardiaca (FC) para ambos grupos fue de 90.34 ± 8.14 ppm, el promedio de saturación de oxigeno (SO2) fue de 95.47 ± el 1.79% y el CO2 espirado fue de 31.13 ± 1.89 mmHg y el promedio de la temperatura (t) para ambos grupos fue de  37.51 ± 0.74oC. La tiempo de recumbencia esternal (TRE) para las cerdas donantes fue de 16.50 ± 4.09 min y el tiempo para pararse (TP) para los receptores fue de 30.70 ± 3.27 min. El propofol se puede considerar como anestésico seguro para el uso en la perfusión continua durante la anestesia. Puesto que no tiene un efecto analgésico se recomienda combinarlo con opioides.

Estandarización del método de la bolsa de aire para su utilización en la evaluación in vivo de sustancias con actividad leishmanicida / Standarization of the air pouch method to be used in the evaluation in vivo of leishmanicide substances

La evaluación de sustancias con actividad Leishmanicida in vivo, actualmente se realiza en modelos animales (ratones o hámster), previo aislamiento y cultivo de los parásitos de Leishmania ssp. y suposterior inoculación y producción de infección en el animal. En este estudio se adaptó la técnica de ®Bolsa de aire¼ descrita por Edwars y colaboradores en 1981, inoculando promastigotes de Leishmania Viannia panamensis (cepa MHOM/CO/87/UA140) vía subcutánea en el espacio interescapular en hámster dorados (Mesocricetus auratus) machos, con el fin de estandarizar un método adecuado para la evaluación in vivo de la actividad leishmanicida de sustancias de origen natural, sintético o hemisintético. En los días 30, 60 y 90 post-inoculación, a cada reactivo biológico se le realizó frotis y aspirado para cultivo en medio NNN para determinar la presencia de infección; en el día 90 a los animales se les aplico la eutanasia, y se tomaron biopsias del sitio de inoculación. Las muestras se procesaron y colorearon con los métodos de Hematoxilina - Eosina y Giemsa. Los aspectos éticos relacionados con el manejo y cuidado de los hámster contaron con la aprobación del ”Comité deÉtica de Animales de Experimentación” de la Universidad de Antioquia y del Ministerio de Salud. La “bolsa de aire” se formo en todos los especímenes, encontrándose permeable, revestida por varias hileras de células simulando el revestimiento articular y a su alrededor se encontraron macrófagos, linfocitos y polimorfonucleares. Sin embargo, no se logro el establecimiento de la infección en los animales en estudio, debido posiblemente a varios factores entre los cuales se puede mencionar la patogenicidad de la cepa del parásito utilizado, la vía y sitio de inoculación y la respuesta inmunologica del Hámster.