An innovative carrier for the formulation of amorphous solid dispersion by hot-melt extrusion with no further downstream processes: a case study with indomethacin.

The aim of this work was to study the possibility to use SepitrapTM as a carrier for the formulation of amorphous solid dispersions by HME (hot melt extrusion) processing aiming solubility enhancement of poorly water-soluble drugs. SepitrapTM is a microencapsulated powder solubilizer designed to simplify the manufacture of drugs in oral solid forms, not yet tested for this purpose. The performance of SepitrapTM was evaluated in HME processing for amorphous solid dispersions of poorly-water soluble drugs with indomethacin as a model drug. The study was conducted using a twin-screw extruder, two compositions of SepitrapTM and different loads of indomethacin, demonstrating that SepitrapTM could represent a new range of carriers for amorphous solid dispersions for HME processing, reducing necessary downstream steps such as grinding.

Comparison study of physicochemical and biopharmaceutics properties of hydrophobic drugs ground by two dry milling processes.

This study focuses on the dry milling of biopharmaceutical classification system (BCS) class II molecules. These molecules have a limited bioavailability because of their low aqueous solubility, poor water wettability and low dissolution rate. In order to improve these properties, indomethacin (IND) and niflumic acid (NIF) were milled using two different types of equipment: Pulverisette 0® and CryoMill®. Milled samples were characterized and compared to commercial molecules. IND shows a modified solid state, like surface crystallinity reduction and an increase in water vapor adsorption from to 2- up to 5-fold due to milling processes. The obtained solubility data resulted in an improvement in solubility up to 1.2-fold and an increase in initial dissolution kinetics: 2% of dissolved drug for original crystals against 25% for milled samples. For NIF no crystallinity reduction, no change of surface properties and no solubility improvement after milling were noticed. In addition, milled particles seemed more agglomerated resulting in no changes in dissolution rate compared to the original drug. IND solubility and dissolution enhancement can be attributed to the modification of surface area, drug crystallinity reduction, and water sorption increase due to specific behavior related to the drug crystal disorder induced by milling process.

Candidemia in a Portuguese tertiary care hospital: Analysis of a 2-year period.

BACKGROUND: Candidemia is a nosocomial infection of increasing importance, associated with high morbidity and mortality. The aim of this study is to describe the species distribution, risk factors, management and outcomes of patients with candidemia. METHODS: We conducted a retrospective study at Centro Hospitalar Universitário de São João, Portugal, between January 2016 and December 2017. RESULTS: A total of 117 candidemia episodes (n=114 patients) were included. Median age was 65 years, with an increased prevalence of older ages. Candida albicans (51.3%) was the most prevalent species, followed by C. glabrata (22.2%), C. parapsilosis (15.4%), C. tropicalis (4.3%) and C. lusitaniae (2.6%). Forty-two patients (35.9%) did not receive antifungal drugs after diagnosis of candidemia. Echinocandins were used as first-line drug therapy in half of the treated patients (50.7%). The median EQUAL Candida Score was 6/17 (IQR 6-9) for patients without central venous catheter (CVC) and 11/20 (IQR 6-14) for patients with CVC. The 30 days-mortality was 31,6% and was not significantly associated with the timing of antifungal therapy and the EQUAL Candida Score. CONCLUSION: The distribution of Candida species has changed in recent years, with an increase in the proportion of C. albicans and C. glabrata. Rapid diagnostic tests, empiric antifungal therapy and source control are essential to improve the prognosis of patients with candidemia. More multicentric prospective studies are needed to evaluate the association of mortality with the timing of antifungal therapy or the EQUAL Candida Score.

Characterization of spermatogonial cells and niche in the scorpion mud turtle (Kinosternon scorpioides).

Undifferentiated spermatogonia (Aund) or spermatogonial stem cells (SSCs) are committed to the establishment and maintenance of spermatogenesis and fertility throughout a male's life and are located in a highly specialized microenvironment called niche that regulates their fate. Although several studies have been developed on SSCs in mammalian testis, little is known about other vertebrate classes. The present study is the first to perform a more detailed investigation on the spermatogonial cells and their niche in a reptilian species. Thus, we characterized Aund/SSCs and evaluated the existence of SSCs niche in the Kinosternon scorpioides, a freshwater turtle found from Mexico to northern and central South America. Our results showed that, in this species, Aund/SSCs exhibited a nuclear morphological pattern similar to those described for other mammalian species already investigated. However, in comparison to other spermatogonial cell types, Aund/SSCs presented the largest nuclear volume in this turtle. Similar to some mammalian and fish species investigated, both GFRA1 and CSF1 receptors were expressed in Aund/SSCs in K. scorpioides. Also, as K. scorpioides Aund/SSCs were preferentially located near blood vessels, it can be suggested that this niche characteristic is a well conserved feature during evolution. Besides being valuable for comparative reproductive biology, our findings represent an important step towards the understanding of SSCs biology and the development of valuable systems/tools for SSCs culture and cryopreservation in turtles. Moreover, we expect that the above-mentioned results will be useful for reproductive biotechnologies as well as for governmental programs aiming at reptilian species conservation.

Duration of spermatogenesis and identification of spermatogonial stem cell markers in a Neotropical catfish, Jundiá (Rhamdia quelen).

Spermatogenesis is a process driven by stem cell, where germ cell cycle is under the control of a specific genotype species. Considering that Jundiá (Rhamdia quelen) is a Neotropical catfish with great economical importance and useful experimental model, little information is available on basic aspects of its reproductive biology, especially on spermatogenesis. As a result, this study aimed to characterize the male germ cells, estimate the duration of spermatogenesis and evaluate the expression of selected stem cell genes in Jundiá testis. Similar to other fish species, our results showed a remarkable decrease of germ cell nuclear volume during Jundiá spermatogenesis, particularly from type A undifferentiated to late type B spermatogonia and from diplotene to late spermatids. Using a S-phase marker, bromodeoxyuridine (BrdU), the combined duration of meiotic and spermiogenic phases in this species was estimated in approximately 7 days. This is considered very short when compared to mammals, where spermatogenesis last from 30 to 74 days. Selected stem cell genes were partially sequenced and characterized in Jundiá testis. Expression analysis showed higher plzf and pou5f3 mRNA levels in the cell fractions enriched by type A undifferentiated spermatogonia. These results were further confirmed by in situ hybridization that showed strong signal of plzf and pou5f3 mRNA in type A undifferentiated spermatogonia. Altogether, these information will expand our knowledge of the reproductive biology of this species, contributing to improve its production and management, and also for biotechnological applications, such as germ cell transplantation.

Metal complexes for multimodal imaging of misfolded protein-related diseases.

Aggregation of misfolded proteins and progressive polymerization of otherwise soluble proteins is a common hallmark of a wide range of highly debilitating and increasingly prevalent diseases, including amyotrophic lateral sclerosis, cerebral amyloid angiopathy, type II diabetes and Parkinson's, Huntington's and Alzheimer's diseases. There is a growing interest in creating imaging agents to detect such aggregates in various imaging modalities, including PET, SPECT and MRI. We present here an overview of recent efforts from the perspective of early diagnosis of amyloid diseases, with a major focus on Aß detection and metal complexes bearing PiB units.

Successful xenogeneic germ cell transplantation from Jundia catfish (Rhamdia quelen) into adult Nile tilapia (Oreochromis niloticus) testes.

Fish germ cell transplantation presents several important potential applications for aquaculture, including the preservation of germplasm from endangered fish species with high genetic and commercial values. Using this technique in studies developed in our laboratory with adult male Nile tilapias (Oreochromis niloticus), all the necessary procedures were successfully established, allowing the production of functional sperm and healthy progeny approximately 2months after allogeneic transplantation. In the present study, we evaluated the viability of the adult Nile tilapia testis to generate sperm after xenogeneic transplant of germ cells from sexually mature Jundia catfish (Rhamdia quelen) that belong to a different taxonomic order. Therefore, in order to investigate at different time-periods post-transplantation, the presence and development of donor PKH26 labeled catfish germ cells were followed in the tilapia seminiferous tubules. From 7 to 20days post-transplantation, only PKH26 labeled spermatogonia were observed, whereas spermatocytes at different stages of development were found at 70days. Germ cell transplantation success and progression of spermatogenesis were indicated by the presence of labeled PKH26 spermatids and sperm on days 90 and 120 post-transplantation, respectively. Confirming the presence of the catfish genetic material in the tilapia testis, all recipient tilapias evaluated (n=8) showed the genetic markers evaluated. Therefore, we demonstrated for the first time that the adult Nile tilapia testis offers the functional conditions for development of spermatogenesis with sperm production from a fish species belonging to a different order, which provides an important new venue for aquaculture advancement.

Functionalized nanomaterials: are they effective to perform gene delivery to difficult-to-transfect cells with no cytotoxicity?

Nanodiamonds (NDs), multiwalled carbon nanotubes (MWCNTs) and gold nanorods (NRs) can be functionalized to promote gene delivery to hard-to-transfect cells with higher transfection efficiency than cationic lipids, and inducing less cell death.

Germ cell transplantation as a potential biotechnological approach to fish reproduction.

Although the use of germ cell transplantation has been relatively well established in mammals, the technique has only been adapted for use in fish after entering the 2000s. During the last decade, several different approaches have been developed for germ cell transplantation in fish using recipients of various ages and life stages, such as blastula-stage embryos, newly hatched larvae and sexually mature specimens. As germ cells can develop into live organisms through maturation and fertilization processes, germ cell transplantation in fish has opened up new avenues of research in reproductive biotechnology and aquaculture. For instance, the use of xenotransplantation in fish has lead to advances in the conservation of endangered species and the production of commercially valuable fish using surrogated recipients. Further, this could also facilitate the engineering of transgenic fish. However, as is the case with mammals, knowledge regarding the basic biology and physiology of germline stem cells in fish remains incomplete, imposing a considerable limitation on the application of germ cell transplantation in fish. Furthering our understanding of germline stem cells would contribute significantly to advances regarding germ cell transplantation in fish.

Post-traumatic stress disorders, mood disorders, and quality of life in transplant recipients with acute liver failure.

OBJECTIVE: The objective of this study was to assess post-traumatic stress disorder (PTSD), depression, anxiety, and quality of life among acute hepatitis patients undergoing liver transplantation. METHOD: Twenty-four patients underwent transplantation due to acute liver failure. After a brief anamnestic interview to gather demographic and clinical information, each participant underwent a psychological assessment using the SF-36 Health Survey (SF-36), Impact of Events Scale-Revised (IES-R), Hospital Anxiety and Depression (HAD) Scale, as well as Beck Depression Inventory (BDI). RESULTS: The average age of the patients was 41.17 years (±17.03); the mean time after transplantation was 13.50 ± 13.80 months; 83% of the subjects were women. Anxiety symptoms were observed in 33.2% of the participants and depression in 16.7% by HAD and 45.8% by BDI. By IES-R 46.2% of patients presented high to severe levels of stress. The majority of participants (54.2%) showed good quality of life scores. A correlation analysis indicated a significant negative association between SF-36 score and other scales (from -0.514 to -0.681). Upon a comparative analysis, a significant difference between groups was only noted in SF-36 score (P = .032). CONCLUSION: The results led us to conclude that anxiety, depression, and PTSD were negatively associated with quality of life in this population. These symptoms also contributed directly to determine well-being among these patients.

Preparation and characterization of carnauba wax nanostructured lipid carriers containing benzophenone-3.

Nanostructured lipid carriers (NLCs) are potential active delivery systems based on mixtures of solid lipids and liquid oil. In this paper, aqueous dispersions of NLCs were prepared by a hot high-pressure homogenization technique using carnauba wax as the solid lipid and isodecyl oleate as the liquid oil. The preparation and stability parameters of benzophenone-3-loaded NLCs have been investigated concerning particle size, zeta potential and loading capacity to encapsulate benzophenone-3, a molecular sunscreen. The current investigation illustrates the effect of the composition of the lipid mixture on the entrapment efficiency, in vitro release and stability of benzophenone-3-loaded in these NLCs. A loading capacity of approximately 5% of benzophenone-3 (m(BZ-3) /m(lipids) ) was characteristic of these systems.

[System of evaluation, application manual and cut-off score for the cube copying test]. / Sistema de evaluación, manual de aplicación y puntuación de corte para la copia del cubo.

INTRODUCTION: One of the difficulties that arises in copying figures in clinical practice and in research is that of how to evaluate performance quantitatively. AIM: To implement a system for evaluating the copying of a solid cube in adults that is sensitive, simple to apply, easily reproducible and which establishes a cut-off score that distinguishes between a well-copied cube and others with alterations. SUBJECTS AND METHODS: The system proposed here is descriptive and is based on the compulsory features a set of nine lines (three groups of three parallel horizontal, vertical and sloping lines) must have in order to form a cube that offers the visual effect of having three dimensions when put together. The elements that are taken into account are the number of faces, the relation between the sizes of the faces, the size of the copy and the characteristics of the groups of horizontal, vertical and sloping lines. The maximum possible score is 9 points (perfect). Four scorers performed a blind analysis of copies of cubes drawn by 215 subjects; copies ranged from normal to others with serious alterations. RESULTS: In all a total of 860 evaluations were obtained (215 x 4). The kappa coefficient was 0.6013 and the Cronbach's alpha coefficient was 0.991. Cubes with a score equal to or above 6 were considered to have been copied correctly. CONCLUSIONS: The system succeeded in evaluating all kinds of cubes and the data set was reliable and consistent. Taking into consideration the extent to which the copies can vary as well as the subjectivity inherent in any clinical method, we believe that the system displayed a high degree of inter-evaluator reliability. The cut-off score can help researchers and doctors with little experience in evaluating the results of the cube copying test.

Neuropsychological assessment in kidney and liver transplantation candidates.

INTRODUCTION: Chronic renal and liver diseases are associated with cognitive and intellectual impairment, which can be irreversible even after kidney or liver transplantation. OBJECTIVE: We sought to investigate the presence of cognitive deficits in organ transplantation candidates. METHODS: From May 2005 to March 2006, 35 organ transplantation candidates, of mean age 46.71 (+/- 13.01) years, 54.3% including females and 7.29 (+/- 4.22) years mean formal schooling. Of those, 27 (77%) were renal and 8 (23%), liver transplantation candidates. All subjects underwent a neuropsychological assessment battery designed to evaluate attention performance, executive functions, memory, language, visuaospatial, and intellectual skills. RESULTS: We found impairments in attention performance (attention span [34.3%], sustained attention [76.5%], and divided attention [77.8%]), executive functions (category formation [58.3%], errors [61.5%], and perseverative errors [30.4%]), memory (working memory [57.1%], verbal [37.1%] and visual short-term memory [31.4%], verbal [25.7%] and visual long-term memory [51.4], verbal learning [42.9%], interference susceptibility [42.9%], and verbal recognition memory [20.6%]), language (comprehension [38.1%], and vocabulary [30.8%]), visuaospatial (45.8%), and intellectual skills (50.0%). CONCLUSION: Neuropsychological (cognitive) deficits in transplant candidates are frequent, regardless of the kind of transplantation. The deficits involve several cognitive skills, such as attentional processes, executive functions, memory, language, visuaospatial, and intellectual skills. Therefore, we concluded that a pretransplant neuropsychological assessment is an important measure to detect impairments and to help understand how these difficulties can interfere with patient self-care before and after transplantation.

Psychosocial profile of patients on the liver transplant list.

INTRODUCTION: Researchers have shown that psychosocial factors influence adherence to treatment and, consequently, prognosis. Psychosocial assessment during the preoperative outpatient phase has allowed physicians to make better treatment decisions. OBJECTIVES: We sought to describe the psychosocial profile of patients on the liver transplant list referred for psychological assessment. METHOD: We reviewed all files of patients referred for pretransplant psychological assessment over a 14-month period. RESULTS: The sample included 73 patients: mean age 49.46 +/- 11.18 years and; mean formal schooling 7.35 +/- 4.65 years. Of those, 56.1% reported a cognitive complaint; 33%, symptoms of anxiety, and 17%, depression. Of the sample, 45.8% were referred for alcohol use, and 37.5% for mood changes. During the assessment, 15.9%, 34.4%, and 1.6%, of patients reported using alcohol, tobacco, or some other type of drug, respectively. Of patients, 53.6% reported having used alcohol at some point in their life-26% tobacco and 10.9% some other type of drug. After the assessment, the main referrals were 35.6% to family counseling, 32.1% to cognitive assessment, and 28.5% to psychological counseling. CONCLUSION: This study highlighted the prevalence of cognitive complaints and current/previous use of chemical substances. These findings were identified through psychological inquiry, which was able to direct the individual treatments. Although these conditions prevailed among this population, their early detection allowed for early intervention to improve adherence and minimize possible intervening problems.

Evaluation of efficacy in a liver pretransplantation orientation group.

BACKGROUND: The medical context recognizes the efficiency of working with groups of patients. Group interventions can intensify the understanding, ability, and notion of recognizing the patient's own condition, increasing the responsibility for him- or herself. This survey sought to evaluate the efficacy of an interdisciplinary orientation group for hepatic transplantation preoperatively. MATERIALS AND METHODS: The opinions of all patients on a waiting list for liver transplantation and their accompanying persons were evaluated from August to December 2005 through a questionnaire with 17 relevant items concerning the transplantation process. The group efficacy was evaluated according to the percentage of correct answers from the subjects before and after attending the group. RESULTS: The results showed a 59% increase in correct answers for the evaluated items after group attendance. The items which showed significant improvement were: what should I do after being called for transplantation; average time of admission to hospital and ICU; use of immunosuppressive drugs; clinical conditions for transplantation; frequency of appointments with the surgeon within the first month; physical activities; diet; blood transfusion; and forgetting medication. A ceiling effect was observed upon reevaluation of the previous conditions for transplantation item. CONCLUSIONS: The percentage of health improvement after attending the group demonstrated an impact of the interdisciplinary orientation intervention on the instruction of patients and their accompanying persons, thus representing an important step in their training process.

Evaluation of efficacy in a pancreas and pancreas-kidney pretransplantation orientation group.

BACKGROUND: There is evidence of benefits from psycho-educational groups in the compliance of patients undergoing complex procedures. Psycho-educational groups provide information, elucidate doubts and realities, fade out fantasies, and help lessen patients' anxieties, thus minimizing the chances of complications or irregular behavior. The objective of this study was to evaluate the efficacy of an interdisciplinary orientation group for pretransplantation preparation for pancreas/pancreas-kidney grafting. MATERIALS AND METHODS: All patients and their accompanying persons who attended information groups from February to August 2005 completed a questionnaire with 15 relevant items about the transplantation process. The efficiency of the orientation group was evaluated according to the percentage of correct answers before and after attending the group. RESULTS: Twenty-seven subjects were evaluated demonstrating an increased number of right answers in 78% of the evaluated items after group attendance. An important improvement was observed in the following items: function of serum sent to the Central Laboratory; serum replacement period; kind of renal donor; blood transfusion; using medicaments; and how often should the patient return for an appointment with the surgeon within the first month. Further items such as surgery risks, using immunosuppressive drugs, and forgetting the medication showed 100% correct answers before and after attending the group. CONCLUSIONS: Results suggest that the pretransplantation orientation group is an efficient way to provide information. Applying a knowledge verification questionnaire before and after the group helps to understand the difficulties of participants, thereby guiding the team and elucidating questions that need more consideration.

Radiolanthanide complexes with tetraazamacrocycles bearing methylphosphonate pendant arms as bone seeking agents.

AIM: Radiolanthanide complexes with ligands bearing phosphonate groups have demonstrated their usefulness as bone seeking agents. Herein, we report on the synthesis of 153Sm and 166Ho complexes with 12- to 14-membered macrocycles containing different number of methylphosphonate pendant arms and their in vitro and in vivo evaluation in order to assess the effect of the cavity size and type of appended arms on their biological behavior. METHODS: Radioactive macrocycle complexes were prepared by reaction of (153)Sm/(166)Ho nitrates with four different tetraazamacrocycles bearing methylphosphonate groups. Radiochemical behavior, in vitro stability and charge of complexes were studied by chromatography and electrophoresis. The lipophilicity, plasmatic protein binding and adsorption onto hydroxyapatite (HA) were evaluated by in vitro assays. Biodistribution was assessed in CD-1 mice. Radiolabeling efficiency depends both on radionuclide and ligand structure. All the complexes are hydrophilic with an overall negative charge and relatively low protein binding. High in vitro stability in human serum and adsorption onto HA was found for all the complexes. RESULTS: Biodistribution and in vivo stability studies have demonstrated promising biological profile for targeted radiotherapy, namely a rapid tissue clearance from most organs and rapid total excretion. Additionally, 166Ho-tritp has a high bone uptake, which led to high bone/ blood and bone/muscle ratios. CONCLUSIONS: Our results clearly demonstrate that 12- and 13-membered macrocyclic ligands led to stable complexes with biological profile adequate to radionuclide therapy. The favorable in vivo behavior highlights the interest to further investigate these or closely related complexes to be used as bone seeking agents.

Phytoplankton nyctemeral variation at a tropical river estuary (Itamaracá-Pernambuco-Brazil).

The Botafogo estuary is of socio-economical importance for Pernambuco State. It is located at the north of Santa Cruz Channel, Itamaracá, Pernambuco, Brazil (07 degrees 42' 50 "S and 34 degrees 52' 10" W). There is a critical need to understand its functioning because of the rate at which this area is being converted to land uses. The phytoplankton dynamics was studied to enhance the knowledge of and verify the possible changes which have occurred in this ecosystem. Sampling was carried out with a plankton net 65 micrometers mesh size for qualitative data, and a Van Dorn bottle for quantitative data. Concurrent hydrological and chlorophyll-a data were collected. Samplings were made in one fixed station in July 1996 (rainy season) and December 1996 (dry season), at 3-hour intervals during 24 hours. Eighty-seven specific and infra-specific taxa were identified from net plankton samples. Diatoms were most frequent, mainly Coscinodiscus centralis and Odontella regia in the rainy and dry seasons, respectively. Phytoplankton density varied from 205,000 to 1,210,000 cell.L-1 in the dry season, and from 230,000 to 2,510,000 cell.L-1 in the rainy season, indicating eutrophic conditions. Most numerically abundant were the diatoms Cyclotella meneghiniana and Cylindrotheca closterium and the phytoflagellates. The ecosystem is polluted, and deleterious effects are minimized by the marine influence which allows periodic water renewal.

Phytoplankton nyctemeral variation at a tropical river estuary (Itamaracá - Pernambuco - Brazil)

O estuário do rio Botafogo tem grande importância socioeconômica para o Estado de Pernambuco. Esse estuário está localizado ao norte do Canal de Santa Cruz, em Itamaracá, Pernambuco, Brasil (07º42'50"S e 34º52'10"W). Há uma necessidade crítica de entender seu funcionamento em razão de a área ser convertida em outros usos. A dinâmica do fitoplâncton foi estudada para aumentar o conhecimento sobre o grupo e verificar as possíveis mudanças ocorridas no ecossistema. Para o estudo qualitativo foram realizadas amostragens com uma rede de plâncton com abertura de malha de 65 micrômetros e, para o quantitativo, utilizou-se uma garrafa de Van Dorn. Paralelamente foram coletados dados de hidrologia e clorofila-a. As amostragens foram feitas em uma estação fixa em julho/1996 (período chuvoso) e dezembro de 1996 (período seco), em intervalos de 3 horas, durante 24 horas. Foram identificados 87 táxons específicos e infra-específicos no plâncton de rede. As diatomáceas foram as mais frequentes, principalmente Coscinodiscus centralis e Odontella regia, respectivamente, nos períodos chuvoso e seco. A densidade fitoplanctônica variou de 205.000 a 1.200.000 céls.Lû1 no período seco, e de 230.000 a 2.510.000 céls.Lû1 no chuvoso, indicando condições eutróficas. A maior abundância numérica foi das diatomáceas Cyclotella meneghiniana e Cylindrotheca closterium e dos fitoflagelados. O ecossistema é poluído e os efeitos negativos são minimizados pela influência marinha, permitindo renovação periódica.