Molecular profiling and feasibility using a comprehensive hybrid capture panel on a consecutive series of non-small-cell lung cancer patients from a single centre.

BACKGROUND: Targeted next-generation sequencing (NGS) is recommended to screen actionable genomic alterations (GAs) in patients with non-small-cell lung cancer (NSCLC). We determined the feasibility to detect actionable GAs using TruSight™ Oncology 500 (TSO500) in 200 consecutive patients with NSCLC. MATERIALS AND METHODS: DNA and RNA were sequenced on an Illumina® NextSeq 550 instrument and processed using the TSO500 Docker pipeline. Clinical actionability was defined within the molecular tumour board following European Society for Medical Oncology (ESMO) guidelines for oncogene-addicted NSCLC. Overall survival (OS) was estimated as per the presence of druggable GAs and treatment with targeted therapy. RESULTS: Most patients were males (69.5%) and former or current smokers (86.5%). Median age was 64 years. The most common histological type and tumour stage were lung adenocarcinoma (81%) and stage IV (64%), respectively. Sequencing was feasible in most patients (93.5%) and actionable GAs were found in 26.5% of patients. A high concordance was observed between single-gene testing and TSO500 NGS panel. Patients harbouring druggable GAs and receiving targeted therapy achieved longer OS compared to patients without druggable GAs. Conversely, patients with druggable GAs not receiving targeted therapy had a trend toward shorter OS compared with driver-negative patients. CONCLUSIONS: Hybrid capture sequencing using TSO500 panel is feasible to analyse clinical samples from patients with NSCLC and is an efficient tool for screening actionable GAs.

Bevacizumab in recurrent ovarian cancer: could it be particularly effective in patients with clear cell carcinoma?

Purpose Treatment of recurrent ovarian carcinoma is a challenge, particularly for the clear cell (CCC) subtype. However, there is a preclinical rationale that these patients could achieve a benefit from antiangiogenic therapy. To assess this hypothesis, we used the growth modulation index (GMI), which represents an intrapatient comparison of two successive progression-free survival (PFS). Methods We conducted a retrospective real-world study performed on 34 patients with recurrent ovarian cancer, treated with bevacizumab-containing regimens from January 2009 to December 2017. The primary endpoint was GMI. An established cut-off > 1.33 was defined as a sign of drug activity. Results 73.5% of patients had high-grade serous ovarian carcinoma (HGSOC), and 17.7% had CCC; 70.6% of patients received carboplatin/gemcitabine/bevacizumab, and 29.4% received weekly paclitaxel/bevacizumab. According to histological subtype, the overall response rate and median PFS were 52% and 14 months for HGSOC and 83.3% and 20 months for CCC, respectively. The overall population median GMI was 0.99; it was 0.95 and 2.36 for HGSOC and CCC, respectively. CCC subtype was significantly correlated with GMI > 1.33 (odds ratio 41.67; 95% confidence interval 3.6–486.94; p = .03). Conclusion Adding bevacizumab to chemotherapy in recurrent CCC is associated with a remarkable benefit in this cohort. The efficacy of antiangiogenic drugs in CCC warrants further prospective evaluation (AU)

Bevacizumab in recurrent ovarian cancer: could it be particularly effective in patients with clear cell carcinoma?

PURPOSE: Treatment of recurrent ovarian carcinoma is a challenge, particularly for the clear cell (CCC) subtype. However, there is a preclinical rationale that these patients could achieve a benefit from antiangiogenic therapy. To assess this hypothesis, we used the growth modulation index (GMI), which represents an intrapatient comparison of two successive progression-free survival (PFS). METHODS: We conducted a retrospective real-world study performed on 34 patients with recurrent ovarian cancer, treated with bevacizumab-containing regimens from January 2009 to December 2017. The primary endpoint was GMI. An established cut-off > 1.33 was defined as a sign of drug activity. RESULTS: 73.5% of patients had high-grade serous ovarian carcinoma (HGSOC), and 17.7% had CCC; 70.6% of patients received carboplatin/gemcitabine/bevacizumab, and 29.4% received weekly paclitaxel/bevacizumab. According to histological subtype, the overall response rate and median PFS were 52% and 14 months for HGSOC and 83.3% and 20 months for CCC, respectively. The overall population median GMI was 0.99; it was 0.95 and 2.36 for HGSOC and CCC, respectively. CCC subtype was significantly correlated with GMI > 1.33 (odds ratio 41.67; 95% confidence interval 3.6-486.94; p = .03). CONCLUSION: Adding bevacizumab to chemotherapy in recurrent CCC is associated with a remarkable benefit in this cohort. The efficacy of antiangiogenic drugs in CCC warrants further prospective evaluation.

Behavioural characterisation of chronic unpredictable stress based on ethologically relevant paradigms in rats.

The chronic unpredictable stress (CUS) paradigm is extensively used in preclinical research. However, CUS exhibits translational inconsistencies, some of them resulting from the use of adult rodents, despite the evidence that vulnerability for many psychiatric disorders accumulates during early life. Here, we assessed the validity of the CUS model by including ethologically-relevant paradigms in juvenile rats. Thus, socially-isolated (SI) rats were submitted to CUS and compared with SI (experiment 1) and group-housed controls (experiment 1 and 2). We found that lower body-weight gain and hyperlocomotion, instead of sucrose consumption and preference, were the best parameters to monitor the progression of CUS, which also affected gene expression and neurotransmitter contents associated with that CUS-related phenotype. The behavioural characterisation after CUS placed locomotion and exploratory activity as the best stress predictors. By employing the exploratory factor analysis, we reduced each behavioural paradigm to few latent variables which clustered into two general domains that strongly predicted the CUS condition: (1) hyper-responsivity to novelty and mild threats, and (2) anxiety/depressive-like response. Altogether, the analyses of observable and latent variables indicate that early-life stress impairs the arousal-inhibition system leading to augmented and persistent responses towards novel, rewarding, and mildly-threatening stimuli, accompanied by lower body-weight gain.

Involvement of midbrain tectum neurokinin-mediated mechanisms in fear and anxiety

Electrical stimulation of midbrain tectum structures, particularly the dorsal periaqueductal gray (dPAG) and inferior colliculus (IC), produces defensive responses, such as freezing and escape behavior. Freezing also ensues after termination of dPAG stimulation (post-stimulation freezing). These defensive reaction responses are critically mediated by γ-aminobutyric acid and 5-hydroxytryptamine mechanisms in the midbrain tectum. Neurokinins (NKs) also play a role in the mediation of dPAG stimulation-evoked fear, but how NK receptors are involved in the global processing and expression of fear at the level of the midbrain tectum is yet unclear. The present study investigated the role of NK-1 receptors in unconditioned defensive behavior induced by electrical stimulation of the dPAG and IC of male Wistar rats. Spantide (100 pmol/0.2 μL), a selective NK-1 antagonist, injected into these midbrain structures had anti-aversive effects on defensive responses and distress ultrasonic vocalizations induced by stimulation of the dPAG but not of the IC. Moreover, intra-dPAG injections of spantide did not influence post-stimulation freezing or alter exploratory behavior in rats subjected to the elevated plus maze. These results suggest that NK-1 receptors are mainly involved in the mediation of defensive behavior organized in the dPAG. Dorsal periaqueductal gray-evoked post-stimulation freezing was not affected by intra-dPAG injections of spantide, suggesting that NK-1-mediated mechanisms are only involved in the output mechanisms of defensive behavior and not involved in the processing of ascending aversive information from the dPAG.

Involvement of midbrain tectum neurokinin-mediated mechanisms in fear and anxiety.

Electrical stimulation of midbrain tectum structures, particularly the dorsal periaqueductal gray (dPAG) and inferior colliculus (IC), produces defensive responses, such as freezing and escape behavior. Freezing also ensues after termination of dPAG stimulation (post-stimulation freezing). These defensive reaction responses are critically mediated by γ-aminobutyric acid and 5-hydroxytryptamine mechanisms in the midbrain tectum. Neurokinins (NKs) also play a role in the mediation of dPAG stimulation-evoked fear, but how NK receptors are involved in the global processing and expression of fear at the level of the midbrain tectum is yet unclear. The present study investigated the role of NK-1 receptors in unconditioned defensive behavior induced by electrical stimulation of the dPAG and IC of male Wistar rats. Spantide (100 pmol/0.2 µL), a selective NK-1 antagonist, injected into these midbrain structures had anti-aversive effects on defensive responses and distress ultrasonic vocalizations induced by stimulation of the dPAG but not of the IC. Moreover, intra-dPAG injections of spantide did not influence post-stimulation freezing or alter exploratory behavior in rats subjected to the elevated plus maze. These results suggest that NK-1 receptors are mainly involved in the mediation of defensive behavior organized in the dPAG. Dorsal periaqueductal gray-evoked post-stimulation freezing was not affected by intra-dPAG injections of spantide, suggesting that NK-1-mediated mechanisms are only involved in the output mechanisms of defensive behavior and not involved in the processing of ascending aversive information from the dPAG.

¿Es necesaria la técnica combinada para la detección del ganglio centinela en el cáncer de mama? / No disponible

Objetivos: Determinar si la punción con colorante mejora los resultados de la biopsia del ganglio centinela. Material y métodos: Ciento cincuenta casos de carcinoma invasor de la mama fueron sometidos a biopsia del ganglio centinela mediante técnica combinada, para determinar si la punción con colorante es rentable. Se realizó inyección subareolar indérmica de una dosis de 0,4 mCi de radioisótopico el día previo a la cirugía. Se practicó gammagrafía en todos los casos. La inyección intraparenquimatosa de 4 cc. de colorante se realizó 20 minutos antes de la cirugía, y se siguió de masaje mamario. Resultados: La tasa de migración fue 92,3% para el radioisótopo y 75% para el colorante (p = 0,01). La media de ganglios resecados fue mayor para la técnica con colorante: 2,6 vs. 1,2 (p = 0,02). No se observaron diferencias en la tasa de falsos negativos (0,2 vs. 0,4) ni en el valor predictivo negativo. La precisión diagnóstica fue mayor para el radioisótopo (90,3% vs. 75% (p = 0,001). El mismo resultado se obtuvo para el porcentaje de éxito técnico (92,3 vs. 75% (0,001)). Conclusiones: La punción con colorante no añade información a la realizada con radioisótopo. A pesar de que la técnica se debe adaptar a las necesidades del centro y a las habilidades del cirujano, una vez superada la curva de validación la técnica radioisotópica por sí misma aporta suficiente información, con menor morbilidad y coste(AU)

Objetives: To determine whether blue dye enhances sentinel node biopsy detection. Material and methods: One hundred fifty hundred consecutive cases of breast cancer were submitted to sentinel node biopsy by combined technique in order to analyze if vital blue was cost-efective. Radioisotope dose was 0,4 mCi of Tc, subareolar intradermic inyection, the day before surgery. Gammagraphy was performed in every case. Colorant was methylene blue, 4 cc administered by intraparenchimatous inyection in upper-outer quadrant 20 minutes previous to surgery, followed by breast massagge. Results: Migration rate was 92.3% for radioisotope and 75% for colorant (p = 0.01). Mean number of nodes excised was higher for colorant: 2,6 vs. 1,2 (p = 0.02). False negative rate showed no difference (0.2 vs. 0.4) nor did negative predictive value. Accuracy to staging (True neg+ true pos./total) was higher for technecium (90.3% vs. 75% (p = 0.001) and so happenned with percentage of technical success (total- no migration): 92.3% vs. 75% (0.001). Conclusions: Colorant did no add any information concerning axillary status. More nodes had to be excised, and bigger incisions were made to achieve direct visualization. Accuracy to stage the lesion and percentage of technical success were higher with radioisotope after the learning curve is achieved and blue injection can be spared(AU)

Adolescent chemical dependency treatment in an HMO.

HMOs are under increasing pressure to expand benefits and services for treatment of adolescents who abuse alcohol and drugs. Little information exists, however, on these programs. This article describes a comprehensive adolescent chemical dependency treatment program within an HMO and presents data on use, costs, and results. Characteristics of clients were similar to adolescents seen in community treatment programs. Less than 1% of the HMO adolescent population sought treatment, and the mean number of outpatient treatment visits was 9.7. The median was between two and three visits, and the mode was one visit. Thirty-four percent of the adolescents required residential treatment, and 65% of the adolescents completed the recommended 28-day stay. The additional premium cost per member per month for adolescent chemical dependency treatment was approximately $0.28. A telephone follow-up survey of a random sample of treated adolescents found that most adolescents had reduced their use of alcohol and drugs and had made improvements in other areas of their lives. Few adolescents, however, met the program's goal of total abstinence.

Characterization of a lectin-like protein isolated from Lachesis muta snake venom.

A lectin-like protein was isolated from L. muta venom by gel filtration on BIO Gel P-100 followed by column Chromatography on DEAE-sephades A-50. The protein eluted at 0.4 M Nacl in 0.01 Tris pH 7.3 and exhibited agglutinin activity toward 0+ human erythrocytes. The protein is a dimer with Mr 28 kDa. Amino acid analysis revealed high content of tryptophan and acid recidues and low content of cysteine and methionine residues. No neutral carbohydrates and sialic acid were detected. Circular dichroic spectrum shows 78% of B structure and 1% of alpha structure. In vitro experiments with erythrocytes from rat, rabbit and dog revealed strong agglutination while red blood cells from mice, sheep and goat were not agglutinated. In vivo experiments using anesthetized rats, a sharp and prolonged fall in the blood pressure was observed at protein dose of 1.5 mg/kg. Double dose of protein caused the death of the animal.

Characterization of a lectin-like protein isolated from Lachesis muta snake venon

A lectin-like protein was isolated from L. muta venom by gel filtration on BIO Gel P-100 followed by column Chromatography on DEAE-Sephadex A-50. The protein eluted at 0.4 M NaCl in 0.01 Tris pH 7.3 and exhibited agglutinin activity toward 0**+ human erytrocytes. The protein is a dimer with Mr 28 kDa. Amino acid analysis revealed high content or tryptophan and acid recidues and low content of cysteine and methionine residues. No neural carbohydrates an sialic acid were detected. Circular dichroic spectrum shows 78% of B structure and 1% of alfa structure. In vitro experiments with ery throcytes from rat, rabbit and dog revealed strong agglutination while red blood cells from mice, sheep and goat were not agglutinated. In vivo experiments using non-anesthetized rats, a sharp and prolonged fall in the blood pressure was observed at protein dose of 1.5 mg/kg. Double dose of protein caused the death of the animal

Effect of chronic administration of Cecropia obtusifolia (Moraceae) on mean arterial pressure in rats

Se le administró un extracto acuoso liofilizado de la morácea Cecropia abtusifolia en el agua de beber a un grupo de ratas (hembras y machos) de la variedad de hipertensas espontáneas. En los machos no se encontraron diferencias significativas en la presión arterial media y en la frecuencia cardiaca al cabo de cuatro semanas de ingerir el extracto. En el caso de las hembras sólo se detectaton diferencias significativas en la presión arterial media, al término de la cuarta semana, pero no en la frecuencia cardíaca

Antihypertensive effect of Cecropia obtusifolia (Moraceae) leaf extract on rats.

A lyophilized aqueous leaf extract of Cecropia obtusifolia proved to be antihypertensive when intravenously administrated to conscious spontaneous hypertensive rats. Forty-five minutes after injection, the maximum fall in arterial pressure (-23.5% relative to preinjection values) was seen and recovery was not complete by the end of the 180 min observation period. The extract was also given to pre-hypertensive SHR and normotensive rats. The fall in blood pressure was more conspicuous in the two SHr groups and was not accompanied by changes in cardiac frequency in any group. This would appear to rule out either a direct or indirect involvement of the heart in regard to the observed hypotension.

Antihypertensive effect of Cecropia obtusifolia (Moraceae) leaf extract on rats

El extracto crudo liofilizado de Cecropia obtusifolia administrado intravenosamente a ratas espontáneamente hipertensas (SHR), no anestesiadas (50 mg/Kg de peso), mostró poseer propiedades antihipertensivas. La presión arterial disminuyó a un mínimo nivel (76.4%) del valor control (161.5 mmHg), 45 minutos después de la infección. El extracto se administró por infusión a tres grupos de animales: hipertensos, prehipertensos (SHR) y a normotensos (Sprague Dawley). La disminución de la presión arterial fue más evidente en el grupo de ratas hipertensas siendo estadísticamente significativa en relación a los otros dos grupos (p, 0.05) de acuerdo con el ANOVA y análisis por el método de Tukey. Esta disminución de la presión arterial media (PAM) no fue acompañada por variaciones de la frecuencia cardiaca en ningún grupo, lo cual indica que no hay participación del nodo sinusal directa o indirectamente en la hipotensión observada

Familial proximal renal tubular acidosis. A distinct clinical entity.

We have studied a family in which nine members present hyperchloremic acidosis with normal plasma creatinine and good ability to acidify urine. Renal functions, other than bicarbonate wasting, are normal, which identifies the condition as a pure form of proximal renal tubular acidosis. The acidosis persists into adult life and appears to be inherited as an autosomal dominant trait. All affected members are asymptomatic and the only peculiar finding is a decrease in stature. No hypercalciuria was detected, and no evidence of rickets or osteomalacia was found by x-ray studies. We consider these findings characteristic of a familial trait different from that in previously reported cases of renal tubular acidosis.

Role of the baroreceptor reflex in the early phases after removing the renal artery constriction in conscious renal hypertensive rats.

Hemodynamic studies in unanesthetized rats with chronic one-kidney-Goldblatt hypertension showed a 25% increase in cardiac output and a 42% increase in peripheral resistance. Removal of renal artery constriction under either anesthesia and minor surgical trauma produced an immediate 20% drop in arterial pressure. At the end of the 6 observation period the pressure dropped 30% but still remained at a moderate hypertensive level. The hemodynamic measurement at that time suggested that the pressure drop was the result of a decrease in cardiac output. However, the data obtained 1 hour after removal of the constriction suggested that a vasodilating mechanism may also contribute to pressure normalization in the early phase of reversal of renal hypertension. In the sham-operated hypertensive rats the pressure remained unchanged, while the cardiac output dropped due to compensation by a proportional increase in peripheral resistance. In contrast, in the unclipped animals the same drop in cardiac output produced an equivalent fall in pressure because no change in peripheral resistance occurred. This was not due to an insufficiency of the baroreceptor reflex since bilateral splanchnicectomy performed at that time produced a striking hypotensive response, indicating an overactivity of the sympathetic system possibly due to the baroreceptor still reset to operate at a hypertensive level.

Role of the baroreceptor reflex in the early phases after removing the renal artery constriction in conscious renal hypertensive rats.

Hemodynamic studies in unanesthetized rats with chronic one-kidney-Goldblatt hypertension showed a 25

increase in cardiac output and a 42

increase in peripheral resistance. Removal of renal artery constriction under either anesthesia and minor surgical trauma produced an immediate 20

drop in arterial pressure. At the end of the 6 observation period the pressure dropped 30

but still remained at a moderate hypertensive level. The hemodynamic measurement at that time suggested that the pressure drop was the result of a decrease in cardiac output. However, the data obtained 1 hour after removal of the constriction suggested that a vasodilating mechanism may also contribute to pressure normalization in the early phase of reversal of renal hypertension. In the sham-operated hypertensive rats the pressure remained unchanged, while the cardiac output dropped due to compensation by a proportional increase in peripheral resistance. In contrast, in the unclipped animals the same drop in cardiac output produced an equivalent fall in pressure because no change in peripheral resistance occurred. This was not due to an insufficiency of the baroreceptor reflex since bilateral splanchnicectomy performed at that time produced a striking hypotensive response, indicating an overactivity of the sympathetic system possibly due to the baroreceptor still reset to operate at a hypertensive level.

Role of the baroreceptor reflex in the early phases after removing the renal artery constriction in conscious renal hypertensive rats

Hemodynamic studies in unanesthetized rats with chronic one-kidney-Goldblatt hypertension showed a 25

increase in cardiac output and a 42

increase in peripheral resistance. Removal of renal artery constriction under either anesthesia and minor surgical trauma produced an immediate 20

drop in arterial pressure. At the end of the 6 observation period the pressure dropped 30

but still remained at a moderate hypertensive level. The hemodynamic measurement at that time suggested that the pressure drop was the result of a decrease in cardiac output. However, the data obtained 1 hour after removal of the constriction suggested that a vasodilating mechanism may also contribute to pressure normalization in the early phase of reversal of renal hypertension. In the sham-operated hypertensive rats the pressure remained unchanged, while the cardiac output dropped due to compensation by a proportional increase in peripheral resistance. In contrast, in the unclipped animals the same drop in cardiac output produced an equivalent fall in pressure because no change in peripheral resistance occurred. This was not due to an insufficiency of the baroreceptor reflex since bilateral splanchnicectomy performed at that time produced a striking hypotensive response, indicating an overactivity of the sympathetic system possibly due to the baroreceptor still reset to operate at a hypertensive level.