Elexacaftor/tezacaftor/ivacaftor, a game-changer in cystic fibrosis: The Portuguese experience.

BACKGROUND: Phase 3 trials of elexacaftor/tezacaftor/ivacaftor (ELX/TEZ/IVA) combination treatment in people with cystic fibrosis (CF) with ≥1 F508del-CFTR allele showed profound short-term effects on lung function, weight, and pulmonary exacerbations (PEx). The authors conducted a 12-month study to add evidence on the real-world long-term effectiveness and safety of CFTR modulator therapy with ELX/TEZ/IVA in Portuguese CF adult population. METHODS: Ambispective, multicentre, observational, real-life study involving all the Portuguese CF Reference Centres. Adult patients on treatment with ELX/TEZ/IVA combination outside clinical trials were included. Demographics, efficacy, and safety variables on the first 12 months of treatment were compared with the pre-treatment year. RESULTS: 132 adult people with CF were included, of which 119 completed 12 months treatment (mean duration of treatment 21.5 months). Mean age was 31.7 ± 11.0 years, 53 % patients were homozygous for the F508del variant, baseline sweat chloride was 86.7 ± 25.9 mmol/L and pre-treatment percent-predicted FEV1 was 77.9 ± 19.7 %. At 1 year, mean absolute change from baseline in FEV1 was +0.46L (95 % CI: 0.37, 0.55; p < 0.001) and +13.9 percentage points (95 % CI: 11.5, 16.2; p < 0.001). PEx episodes decreased by 78 % (p < 0.001) and hospitalizations for PEx decreased by 91.4 % (p < 0.001). Body mass index (BMI) increased 1.2 kg/m2 (95 % CI: 0.9, 1.5; p < 0.001). Mean sweat chloride variation was -44.5 mmol/L (95 % CI: -49.8, -39.2; p < 0.001). No correlation was found between sweat chloride and lung function (r = -0.116, p = 0.335). There were no major safety concerns. Of note, headache was reported in 7.6 % and neuropsychiatric manifestations occurred in 12.6 % treated patients, being anxiety and depressive disorders the most common. CONCLUSIONS: ELX/TEZ/IVA treatment in Portuguese adults with CF was associated with significant improvement in lung function, a drop in PEx and PEx-related hospitalizations and increase in BMI at 12 months and was well tolerated. These results add knowledge to our understanding of clinical benefits and tolerability of ELX/TEZ/IVA. Careful evaluation of adverse effects of ELX/TEZ/IVA therapy and its determinants, mainly concerning mental health, are a research priority.

Double blind randomized clinical trial comparing minimally- invasive envelope flap and conventional envelope flap on impacted lower third molar surgery.

BACKGROUND: The latest trend in surgery is to look for minimally invasive procedures, with fewer complications and a shorter recovery time. This study aims to compare the minimally- invasive envelope flap, with smaller incision and fewer dissection and the conventional envelope flap, with a 20mm incision, on impacted mandibular third molar surgery, focusing on the hypothesis that there were no differences in postoperative outcomes. MATERIAL AND METHODS: A double-blind randomized clinical trial was designed to compare both incisions, focused on determining the approach with minor postoperative side-effects and minor impact on quality-of-life. A total of 60 patients were enrolled for the study if their presented impacted mandibular third molar and was 18-years-old or more. Both groups were evaluated from time elapsed on the surgery, maximum mouth opening, swelling and quality of life assessment. RESULTS: The flap choice influenced facial swelling (p=0,03), pain on the first three days (p=0,037), interference with oral hygiene (p=0,019) and discomfort on speech (p=0,07). Chewing, swallowing, trismus, pain after seven days, postoperative complications and other quality-of-life arrangements were no different between groups. CONCLUSIONS: The minimally- invasive envelope flap could lead to a less painful experience for the patient, with fewer impact on the oral hygiene and speech discomfort.

Matteucinol combined with temozolomide inhibits glioblastoma proliferation, invasion, and progression: an in vitro, in silico, and in vivo study.

Glioblastoma is the most prevalent and malignant brain tumor identified in adults. Surgical resection followed by radiotherapy and chemotherapy, mainly with temozolomide (TMZ), is the chosen treatment for this type of tumor. However, the average survival of patients is around 15 months. Novel approaches to glioblastoma treatment are greatly needed. Here, we aimed to investigate the anti-glioblastoma effect of the combination of matteucinol (Mat) (dihydroxyflavanone derived from Miconia chamissois Naudin) with the chemotherapeutic TMZ in vitro using tumor (U-251MG) and normal astrocyte (NHA) cell lines and in vivo using the chick embryo chorioallantoic membrane (CAM) assay. The combination was cytotoxic and selective for tumor cells (28 µg/mL Mat and 9.71 µg/mL TMZ). Additionally, the combination did not alter cell adhesion but caused morphological changes characteristic of apoptosis in vitro. Notably, the combination was also able to reduce tumor growth in the chick embryo model (CAM assay). The docking results showed that Mat was the best ligand to the cell death membrane receptor TNFR1 and to TNFR1/TMZ complex, suggesting that these two molecules may be working together increasing their potential. In conclusion, Mat-TMZ can be a good candidate for pharmacokinetic studies in view of clinical use for the treatment of glioblastoma.

Matteucinol combined with temozolomide inhibits glioblastoma proliferation, invasion, and progression: an in vitro, in silico, and in vivo study

Glioblastoma is the most prevalent and malignant brain tumor identified in adults. Surgical resection followed by radiotherapy and chemotherapy, mainly with temozolomide (TMZ), is the chosen treatment for this type of tumor. However, the average survival of patients is around 15 months. Novel approaches to glioblastoma treatment are greatly needed. Here, we aimed to investigate the anti-glioblastoma effect of the combination of matteucinol (Mat) (dihydroxyflavanone derived from Miconia chamissois Naudin) with the chemotherapeutic TMZ in vitro using tumor (U-251MG) and normal astrocyte (NHA) cell lines and in vivo using the chick embryo chorioallantoic membrane (CAM) assay. The combination was cytotoxic and selective for tumor cells (28 μg/mL Mat and 9.71 μg/mL TMZ). Additionally, the combination did not alter cell adhesion but caused morphological changes characteristic of apoptosis in vitro. Notably, the combination was also able to reduce tumor growth in the chick embryo model (CAM assay). The docking results showed that Mat was the best ligand to the cell death membrane receptor TNFR1 and to TNFR1/TMZ complex, suggesting that these two molecules may be working together increasing their potential. In conclusion, Mat-TMZ can be a good candidate for pharmacokinetic studies in view of clinical use for the treatment of glioblastoma.

Comparação de métodos para avaliação de beta-hidroxibutirato em ovelhas / Comparison of methods for evaluation of beta-hydroxybutyrate in ewes

Atualmente o uso de sensores portáteis para mensuração de corpos cetônicos está padronizado e difundido na rotina clínica, contudo estudos em ovinos são escassos. Assim, a presente pesquisa objetivou avaliar a acurácia dos sensores portáteis de uso humano e de uso veterinário para a determinação de beta-hidroxibutirato (BHB) em ovelhas no final da gestação e no pós-parto recente. Foram utilizadas 37 amostras de sangue provenientes de nove ovelhas mestiças Corriedale. A determinação bioquímica de BHB no soro, considerada como o padrão-ouro, foi realizada utilizando-se metodologia enzimática colorimétrica. A média obtida na bioquímica sérica foi de 0,497mmol/L; no sensor de uso humano, a média foi igual a 0,537mmol/L, enquanto no sensor de uso veterinário foi de 0,751mmol/L. Foi verificada alta correlação entre o dosímetro de uso humano e o padrão-ouro (r=0,93, P<0,001). A média do aparelho de uso veterinário diferiu das demais (51%; P<0,05), superestimando os resultados em ovelhas. As medições obtidas no aparelho veterinário também apresentaram menor precisão e veracidade. Concluiu-se que o sensor portátil de uso humano é mais acurado e mais preciso no diagnóstico precoce de toxemia da gestação em ovelhas.

Currently the use of portable sensors for measuring ketone bodies is standardized and diffused in the clinical routine, however, studies in sheep are scarce. Therefore, the present study aimed to evaluate the accuracy of the human portable sensor and the veterinary portable sensor for the determination of beta-hydroxybutyrate (BHB) in sheep at the end of gestation and postpartum. We used 37 samples of blood from nine crossbred Corriedale sheep. Biochemical determination of serum BHB, considered gold standard, was performed using colorimetric enzymatic methodology. The mean serum biochemistry was 0.497mmol/L, in the human sensor the mean was 0.537mmol/L, while in the veterinary sensor it was 0.751mmol/L. A high correlation was verified between the dosimeter for human use and the gold standard (r= 0.93, P< 0.001). The mean of the veterinary apparatus differed from the others, being 51% (P< 0,05), higher than the standard, that is, it was less accurate and had lower veracity, overestimating the results in sheep. It was concluded that the portable sensor for human use is more accurate and accurate in the early diagnosis of toxemia of pregnancy in sheep.

Therapeutic nanosystems for oncology nanomedicine.

Cancer is a complex disease which includes many pathologies featuring abnormal growth of healthy cells, invasion and metastization. Cellular uncontrolled growth increases the amount of abnormal cells, these cells reach the blood stream and other healthy tissues giving rise to secondary tumors. Even though great progresses have been made in understanding cancer aetiology and in the development of new anticancer drugs, cancer still remains one of the leading causes of death worldwide. Most common cancers include breast cancer, lung cancer, colorectal cancer and brain cancer. Conventional chemotherapeutics have proven to be inefficient in cancer treatment due to lack of specificity and poor drug accumulation in tumors. In addition, they cause severe side effects. Available treatments must be prolonged on time to achieve some therapeutic effect; however, this often leads to the development of multidrug resistance by tumor cells. Nanotechnology platforms are, therefore, being exploited as potential alternatives. Nanosystems have been reported to target and deliver the drug in situ to selectively kill cancer cells, decreasing toxicity on healthy organs and tissues as well as side effects. Furthermore, some nanosystems have been reported to overcome tumor resistance, at least to some extent. Over the years several nanosystems have been proposed to diagnose and treat cancers, such as dendrimers, polymeric micelles, superparamagnetic iron oxide cores, gold nanoparticles, liposomes and other lipid nanoparticles. Due to their small size and biocompatibility, they can reach the target site without being detected by the immune system and suffer cellular uptake or deliver the drug in the tumor vicinity.

Listeria monocytogenes in HIV-infected patients in a Hospital of Nova Iguaçu, Rio de Janeiro, Brazil

A survey was carried out in a hospital of Nova Iguaçu, Rio de Janeiro, Brazil, in the period from July 1999 to March 2002, to determine the infection rate of Listeria monocytogenes in HIV+ patients with diarrhea symptoms; 134 samples were processed by microbiological methods. The results demonstrated 12.68% of positive samples. However, no statistical differences were observed for age or sex in the studied group, suggesting that this microorganism should be regarded in the differential diagnosis of infectious processes in HIV+ patients in the area

The involvement of the chemokine receptor CXCR2 in neutrophil recruitment in LPS-induced inflammation and in Mycobacterium avium infection.

Knockout mice for CXC receptor 2 (CXCR2) chemokine receptor were used to study the recruitment of neutrophils during acute and chronic inflammatory responses. When treated with lipopolysaccharide (LPS), either intraperitoneally or intratracheally, these animals had a significant reduction in the neutrophil recruitment in the first 24-48 h as compared with control mice. During 15 days of intraperitoneal infection by Mycobacterium avium, the knockout mice showed significantly reduced numbers of neutrophils in the peritoneal cavity as compared with the control mice. In contrast, the recruitment of neutrophils to the lungs during an aerogenic M. avium infection was not affected by the CXCR2 mutation throughout the 60 days of the study. Finally, we could not find any impact of the mutation on the mycobacterial growth of the infected animals. These findings indicate that CXCR2 may be essentially involved in acute inflammatory responses where an early and rapid recruitment of neutrophils is observed.

Effects of recombinant granulocyte-colony stimulating factor administration during Mycobacterium avium infection in mice.

Granulocyte colony-stimulating factor (G-CSF) administration in vivo has been shown to improve the defence mechanisms against infection by different microbes. Here we evaluated a possible protective role of this molecule in a mouse model of mycobacterial infection. The administration of recombinant G-CSF promoted an extensive blood neutrophilia but failed to improve the course of Mycobacterium avium infection in C57Bl/6 or beige mice. G-CSF administration also failed to improve the efficacy of a triple chemotherapeutic regimen (clarithromycin + ethambutol + rifabutin). G-CSF treatment did not protect interleukin-10 gene disrupted mice infected with M. avium. Spleen cells from infected mice treated with G-CSF had a decreased priming for antigen-specific production of interferon gamma compared to control infected mice. Our data do not substantiate previous reports on the protective activity of G-CSF in antimycobacterial immunity using mouse models.

Modulation of neutrophil influx with cell adhesion molecule specific antibodies during nonspecific and immune mediated inflammatory reactions.

Neutrophils are essential for the host defence against infection. However, neutrophils may also mediate damage namely during immune mediated pathologies. We therefore tested whether targeting of different cell adhesion molecules with specific monoclonal antibodies might reduce immune mediated neutrophil recruitment but spare the nonspecific accumulation of neutrophils that is essential for the resistance against acute infections. Neutrophil recruitment was induced by either intraperitoneal injection of casein as a nonspecific phlogistic agent or by i.p. injection of antigen in Mycobacterium bovis Bacille Calmette-Guérin (BCG) immune mice. Similar degrees of inhibition of neutrophil accumulation were observed in both models of inflammation with antibodies directed at CD11a, ICAM-1 and CD11b with the latter showing the most marked effects. Individual targeting of selectins was without effect in immune mediated responses whereas targeting of L or E selectin inhibited nonspecific recruitment of neutrophils. This was apparently not owing to a dosage effect nor to a kinetic difference. The inhibitory effect of anti-CD11b antibodies was most likely as a result of activation of circulating neutrophils rather than the blocking of receptor-ligand interactions. We were therefore unable to selectively abrogate immune mediated neutrophil recruitment with the use of the antibodies selected in this study.

Resistance of virulent Mycobacterium avium to gamma interferon-mediated antimicrobial activity suggests additional signals for induction of mycobacteriostasis.

The cytokine gamma interferon (IFN-gamma) plays a major role in the control of Mycobacterium avium infections. We assessed whether the progressive growth of virulent strains of M. avium was associated with alterations in the production of this cytokine as evaluated by reverse transcription-PCR and detection of immunoreactive cytokine in the serum and in spleen homogenates. We found that IFN-gamma was induced during infection by a virulent strain of M. avium to similar or even higher extents than the levels found during infections by a less virulent strain whose growth was controlled. IFN-gamma produced during infection by both mycobacterial strains was partly derived from T cells and led to activation of macrophages, namely, those that were infected. Concomitant with the development of the infection with the virulent strain of M. avium there was an extensive depletion of lymphocytes in the spleen. Thymectomy alone promoted the proliferation of the virulent, but not of the less virulent, strain of M. avium. Our data indicate that virulent strains of M. avium resist the antimicrobial mechanisms of IFN-gamma-activated macrophages and raise the possibility that a second, T-cell-dependent signal is required for the effective control of mycobacterial replication inside macrophages.

Evaluation of esophageal motility in laryngectomized patients.

Laryngectomy for treatment of laryngeal-pharyngeal carcinomas may impair the sensation in the larynx and epiglottis, with consequent impairment of esophageal motility. Our aim in the present study was to investigate the esophageal motility of laryngectomized patients. Esophageal manometry was performed on 17 patients submitted to laryngectomy 2 to 71 months (median 29 months) before the examination. Eleven were rehabilitated with esophageal voice and six could not speak. Ten swallows of a 5 ml bolus of water were recorded at the lower esophageal sphincter and at 5, 10 and 15 cm above it. The lower esophageal sphincter pressure was measured by the rapid pull-through method and the upper esophageal sphincter pressure by the station pull-through method. The results were compared with those obtained for a control group of 40 healthy volunteers. The amplitude of contractions was lower and the number of nonperistaltic contractions was higher in laryngectomized patients than in volunteers (P < 0.05). The duration of lower esophageal sphincter relaxation (7.4 +/- 1.5 s) was shorter in laryngectomized patients than in volunteers (8.8 +/- 1.6 s, P < 0.05). The upper esophageal sphincter pressure was lower (34.9 +/- 29.1 mm Hg) in laryngectomized patients than in volunteers (61.2 +/- 20.8 mm Hg, P < 0.05). There was no difference between groups in contraction duration or velocity, in the numbers of multipeaked or failed contractions, lower esophageal sphincter pressure or in the number of swallows followed by complete lower esophageal sphincter relaxation. In conclusion, laryngectomy causes esophageal motility impairment characterized by low contraction amplitude, nonperistaltic contraction and shorter lower esophageal sphincter relaxation duration.

Antinociceptive effect of intrathecal neostigmine evaluated in rats by two different pain models

The analgessic efficacy of cholinergic agonists and anticholinesterase agents has been widely recognized. The analgesic effect obtained by activating cholinergic mechanisms, however, seems to depend on the experimental pain model utilized for its evaluation. The antinociceptive effect of intraspinal neostigmine was examined in rats submitted concurrently to the tail flick and formalin tests. Neostigmine (8.25 and 16.5 nmol) produced a dose-dependent antinociceptive effect in the tail flick test (a model of phasic pain) and reduced the first phase (phasic pain) of the animal response to formalin also in a dose-dependent manner. The second phase (tonic pain) of the response to formalin, however, was slightly reduced after a longer period of time only by the higher dose of the anticholinesterase. The effect of neostigmine was not significantly different when the drug was injected into rats submitted exclusively to the tail flick test. The second phase of the animal response to formalin was slightly reduced by neostigmine (8.25 nmol) and strongly inhibited by the higher dose of the anticholinesterase when injection was made after the first phase. We conclude that phasic and tonic pain can both be controlled by high doses of neostigmine. In addition, we show that inhibition by a lower dose of neostigmine of the formalin-induced phasic pain did not prevent the subsequent occurrence of tonic pain produced by the irritant.

Antinociceptive effect of intrathecal neostigmine evaluated in rats by two different pain models.

The analgesic efficacy of cholinergic agonists and anticholinesterase agents has been widely recognized. The analgesic effect obtained by activating cholinergic mechanisms, however, seems to depend on the experimental pain model utilized for its evaluation. The antinociceptive effect of intraspinal neostigmine was examined in rats submitted concurrently to the tail flick and formalin tests. Neostigmine (8.25 and 16.5 nmol) produced a dose-dependent antinociceptive effect in the tail flick test (a model of phasic pain) and reduced the first phase (phasic pain) of the animal response to formalin also in a dose-dependent manner. The second phase (tonic pain) of the response to formalin, however, was slightly reduced after a longer period of time only by the higher dose of the anticholinesterase. The effect of neostigmine was not significantly different when the drug was injected into rats submitted exclusively to the tail flick test. The second phase of the animal response to formalin was slightly reduced by neostigmine (8.25 nmol) and strongly inhibited by the higher dose of the anticholinesterase when injection was made after the first phase. We conclude that phasic and tonic pain can both be controlled by high doses of neostigmine. In addition, we show that inhibition by a lower dose of neostigmine of the formalin-induced phasic pain did not prevent the subsequent occurrence of tonic pain produced by the irritant.

Segregation index: an orsametric for measuring discrimination against minorities.

Orsametrics, the science of data and measurement appropriate to decision-making models, was proposed by H. M. Wagner in 1971. This paper proposes an orsametric measuring the distance between the current distribution of minorities in the organization and the worst distribution that minorities could have, given a specified proportion of minorities present in the organization. Measurement of this distance via the absolute value norm leads to a constrained model in which a convex, absolute-value functional must be maximized. This is accomplished by means of an algorithm designed to take advantage of the special structural properties of the model. An example is provided to demonstrate how the segregation index can be used to track discrimination levels within an organization through time. (Orsametrics: public sector applications-quantification of discriminatory hiring promotion practices; mathematical modelling-nonlinear, maximizing a convex function subject to linear constraints.