Functional chromophores synthesized via multicomponent Reactions: A review on their use as cell-imaging probes.

This review aims to provide a comprehensive overview of recent advancements and applications of fluorescence imaging probes synthesized via MCRs (multicomponent reactions). These probes, also known as functional chromophores, belong to a currently investigated class of fluorophores that are presently being successfully applied in bioimaging experiments, especially in various living cell lineages. We describe some of the MCRs that have been employed in the synthesis of these probes and explore their applications in biological imaging, with an emphasis on cellular imaging. The review also discusses the challenges and future perspectives in the field, particularly considering the potential impact of MCR-based fluorescence imaging probes on advancing this field of research in the coming years. Considering that this area of research is relatively new and nearly a decade has passed since the first publication, this review also provides a historical perspective on this class of fluorophores, highlighting the pioneering works published between 2011 and 2016.

Posterior Circulation Endovascular Thrombectomy for Large-Vessel Occlusion: Predictors of Favorable Clinical Outcome and Analysis of First-Pass Effect.

BACKGROUND AND PURPOSE: Successful vessel recanalization in posterior circulation large-vessel occlusion is considered crucial, though the evidence of clinical usefulness, compared with the anterior circulation, is not still determined. The aim of this study was to evaluate predictors of favorable clinical outcome and to analyze the effect of first-pass thrombectomy. MATERIALS AND METHODS: A retrospective, multicenter, observational study was conducted in 10 high-volume stroke centers in Europe, including the period from January 2016 to July 2019. Only patients with an acute basilar artery occlusion or a single, dominant vertebral artery occlusion ("functional" basilar artery occlusion) who had a 3-month mRS were included. Clinical, procedural, and radiologic data were evaluated, and the association between these parameters and both the functional outcome and the first-pass effect was assessed. RESULTS: A total of 191 patients were included. A lower baseline NIHSS score (adjusted OR, 0.77; 95% CI, 0.61-0.96; P = .025) and higher baseline MR imaging posterior circulation ASPECTS (adjusted OR, 3.01; 95% CI, 1.03-8.76; P = .043) were predictors of better outcomes. The use of large-bore catheters (adjusted OR, 2.25; 95% CI, 1.08-4.67; P = .030) was a positive predictor of successful reperfusion at first-pass, while the use of a combined technique was a negative predictor (adjusted OR, 0.26; 95% CI, 0.09-0.76; P = .014). CONCLUSIONS: The analysis of our retrospective series demonstrates that a lower baseline NIHSS score and a higher MR imaging posterior circulation ASPECTS were predictors of good clinical outcome. The use of large-bore catheters was a positive predictor of first-pass modified TICI 2b/3; the use of a combined technique was a negative predictor.

A ten-color tube with dried antibody reagents for the screening of hematological malignancies.

INTRODUCTION: The workflow in clinical flow cytometry laboratories must constantly be reviewed to develop technical procedures that improve quality and productivity and reduce costs. Using the Beckman Coulter dry coating technology, we customized a ten-color tube with dried antibody reagents, designated the Duraclone screening tube (DST), for screening hematological malignancies. Here, we compared the applicability, clinical and numerical equivalence, and cost and time required for the technical procedures between the liquid reagents and the DST. METHODS: The DST contains CD4 + Kappa-FITC, CD8 + Lambda-PE, CD3 + CD14-ECD, CD33-PE-Cy5.5, CD20 + CD56-PE-Cy7, CD34-APC, CD19-APC-AlexaFluor700, CD10-APC-AlexaFluor750, CD5-Pacific Blue, and CD45-Krome Orange. We evaluated 20 bone marrow samples, 13 peripheral blood samples, 6 lymph node biopsy samples, 5 fine-needle aspirate samples, 5 cerebrospinal fluid samples, and 1 pleural fluid sample. RESULTS: The DST was useful for more than 60% of our samples. It was able to enumerate the majority of the populations in all types of samples with a statistically acceptable correlation with the liquid reagents. The use of the DST translated into significant time and cost savings of 15.8% and 12.3%, respectively, compared with the use of the liquid reagent. The cost was reduced by $14.36 per sample. CONCLUSIONS: The DST is an efficient solution for screening hematological malignancies with improved quality, productivity, standardization, and sustainability. These improvements could benefit patients by providing faster diagnoses using a higher quality and lower cost reagent.

Hemograma e proteínas de fase aguda de bezerros sadios do nascimento aos 30 dias de idade / Hemogram and acute phase proteins of healthy calves from birth to one month old

O conhecimento da dinâmica das alterações nos parâmetros hematológicos e na cinética das proteínas de fase aguda em animais saudáveis nas primeiras semanas de vida é essencial para a interpretação correta dessas avaliações em situações de morbidez e para diferenciar animais sadios e enfermos de forma confiável. Com o intuito de avaliar a cinética desses parâmetros no primeiro mês de vida de bezerros de corte sadios, filhos de vacas primíparas ou pluríparas, amostras de sangue foram coletadas antes da ingestão de colostro e 1, 2, 7, 15 e 30 dias após o nascimento. Os parâmetros eritrocitários foram influenciados pelo número de partos das vacas e o leucograma mostrou alterações características de influência do cortisol fetal liberado por ocasião do nascimento. O teor sérico de proteína total aumentou significativamente após a ingestão do colostro. As concentrações de ceruloplasmina, haptoglobina e proteínas de pesos moleculares 33 kDa e 23 kDa aumentaram significativamente no primeiro dia de vida, seja pela resposta ao nascimento ou pela ingestão do colostro, enquanto os teores de transferrina, albumina e α1-glicoproteína ácida mantiveram-se relativamente estáveis nos primeiros dias de vida, aumentando gradualmente até os 30 dias de idade.

The knowledge of the dynamic in changes of hematologic parameters and the acute phase proteins kinetics in healthy animals in the first weeks of life is essential for the accurate interpretation of these evaluations in morbidity situations, and to reliably differentiate healthy from sick animals. The aim of the study was to evaluate the dynamic of these parameters in the first month of life of healthy beef calves, born from primiparous or multiparous cows, and so blood samples were collected before colostrum intake and 1, 2, 7, 15 and 30 days thereafter to evaluate hemogram and serum proteinogram. The red cell parameters were influenced by the parturition number of cows, and the leukogram showed characteristic changes of the release of fetal cortisol at birth. The total protein level significantly increased after colostrum intake. The concentrations of acute phase proteins ceruloplasmin, haptoglobin and proteins of molecular weight 33 kDa and 23 kDa significantly increased in the first day of life, influenced either by response to birth or colostrum intake, while the levels of transferrin, albumin and α1-acid glycoprotein were relatively stable in the first days, increasing gradually until 30 days of life.

Passive immunity transfer and serum constituents of crossbred calves / Transferência de imunidade passiva e constituintes séricos de bezerros mestiços

Passive immunity transfer (PIT) evaluation is an essential tool for the maintenance of healthy calves during the first months of life. Since lactation number and breed have been proven to influence immunoglobulin levels in colostrum, the aim of this study was to evaluate PIT from primiparous and multiparous Canchim cows to their calves. Blood samples were collected from the calves before colostrum intake and 1, 2, 7, 15 and 30 days thereafter, while colostrum samples from the cows were taken immediately after parturition. Activities of gamma-glutamyl transferase (GGT), alkaline phosphatase (ALP), and concentrations of total protein, albumin, globulins, immunoglobulin A (IgA), immunoglobulin G (IgG), total and ionized calcium, inorganic phosphorus, magnesium, sodium and potassium were evaluated in calves' serum and activities of GGT and ALP and concentrations of total protein, IgA and IgG were assessed in cow's colostrum whey. Immunoglobulins concentrations were evaluated by electrophoresis in polyacrylamide gels. Serum biochemistry evaluations revealed an increase in gamma-glutamyl transferase and alkaline phosphatase activities and in total protein, globulins, immunoglobulin A and immunoglobulin G levels in calves' serum after colostrum intake. Only total protein and light chain immunoglobulin G levels in colostrum whey were affected by the cows' lactation number. Phosphorus and magnesium levels in blood serum increased after colostrum intake, while sodium and potassium levels oscillated in the experimental period. PIT was influenced by the cows' lactation number but was efficient in both groups.

A avaliação da transferência de imunidade passiva (TIP) é uma ferramenta essencial para manutenção de bezerros saudáveis nos primeiros meses de vida. Uma vez que há influência do número de lactações e da raça de vacas nos teores de imunoglobulinas do colostro, o presente estudo foi conduzido com o objetivo de avaliar a TIP de vacas Canchim primíparas e pluríparas aos seus bezerros. Amostras de sangue dos bezerros foram coletadas antes da ingestão de colostro e 1, 2, 7, 15 e 30 dias após o nascimento e amostras de colostro das vacas foram coletadas imediatamente após o parto. As atividades de gamaglutamiltransferase (GGT), fosfatase alcalina (ALP) e as concentrações de proteína total, albumina, globulinas, imunoglobulina A (IgA) e imunoglobulina G (IgG), cálcio total e ionizado, fósforo, magnésio, sódio e potássio foram avaliadas no soro dos bezerros e as atividades de GGT e ALP e as concentrações de proteína total, IgA e IgG foram avaliadas no soro colostral. A concentração de imunoglobulinas foi avaliada por meio de eletroforese em gel de poliacrilamida. As avaliações bioquímicas do soro sanguíneo dos bezerros revelaram aumento nas atividades das enzimas gamaglutamiltransferase e fosfatase alcalina e nos teores de proteína total, globulinas, imunoglobulina A e imunoglobulina G após a ingestão do colostro. Apenas os teores de proteína total e imunoglobulina G de cadeia leve no soro colostral foram influenciados pelo número de lactações das vacas. Os teores de fósforo e magnésio aumentaram após a ingestão de colostro, enquanto as concentrações de sódio e potássio oscilaram no decorrer do período experimental. A TIP foi influenciada pelo número de lactações das vacas, no entanto mostrou-se eficiente em ambos os grupos.

Charge-tagged acetate ligands as mass spectrometry probes for metal complexes investigations: applications in Suzuki and Heck phosphine-free reactions.

An acetate anion bearing an imidazolium cation as its charge tag was reacted with M(OAc)(2) complexes (where M = Ni, Cu, and Pd; in situ reaction) to form members of a new class of charge-tagged metal complexes. The formation of these unprecedented precatalysts with potential for cross-coupling reactions was confirmed by electrospray ionization (and tandem) mass spectrometry. The catalytic performance of the palladium complex was tested in Heck and Suzuki cross-coupling reactions, often with superior activity and yields as compared with Pd(OAc)(2).

Modic changes: anatomy, pathophysiology and clinical correlation.

Studying discovertebral complex anatomy is extremely important for the understanding of the pathophysiology of disc degeneration which leads to vertebral endplates signal changes, also known as Modic changes.The sequelae of disc degeneration are among the leading causes of functional incapacity in both sexes and are one of the most common sources of chronic disability in the working years. Even if the presence of degenerative changes in MRI of the spine is by no means an indicator of symptoms, we are concordant in a positive association between Modic changes and low back pain, above all as a relatively specific but insensitive sign of discogenic low back pain.

Treatment of symptomatic lumbar spinal degenerative pathologies by means of combined conservative biochemical treatments.

Research in spine surgery has proposed new soft and less invasive techniques. These are the results of our experience with oxygen-ozone therapy, which we could experiment within the Italian National Health System over 3 years. A total of 1,920 patients were admitted on the basis of unselected enrolment because of lumbosciatic pain. Patients were divided into three groups: (A) Patients with degenerative disc disease and arthropathy: 509 (26.5%), (B) Patients with failed back surgery syndrome (FBSS): 1,027 (53.489%), and (C) Patients with pure herniated lumbar disc: 384 (20%). The rationale of the treatment for all these different pathologies we have taken into consideration is the biochemical mechanism by which they can engender pain and dysfunction. Treatment for group A: paravertebral injection and phleboclysis (two cycles of 6 sessions, one each 3 days) +endoscopic neurolysis. Treatment for group B: paravertebral injection and phleboclysis (two cycles of 6 sessions, one each 3 days) + endoscopic neurolysis with intradiscal procedure (named percutaneous peridurodiscolysis). Treatment for group C: paravertebral injection (two cycles of 6 sessions, one each 3 days) + percutaneous discolysis.The perceived quality of result for this minimally invasive procedure makes oxygen-ozone therapy an interesting weapon in the hands of doctors. Furthermore, if the technique loses its clinical effectiveness, it can be repeated without harm for the patient, and costs for the health organization are notably very low, above all if compared to surgical procedures.We underline the need that this treatment should be performed in protected structures, in operative rooms, under anesthesiologic control, and in the hands of specialists.

Considerations on the treatment of anterior interosseous nerve syndrome.

Anterior interosseous syndrome (Kiloh-Nevin syndrome) refers to that constellation of signs and symptoms referable to weakness of the pronator quadratus, the flexor pollicis longus, and the flexor digitorum profundus to the index finger.We present our series of 9 patients, affected by AIN Syndrome, and a group of 4 patients affected by pseudo-AIN neuropathies.In the literature there is considerable controversy concerning the treatment, but we agree that understanding of anatomical variants of innervation combined with a thorough physical examination can provide important clues as to where pathology resides. Proper treatment needs a precise and accurate diagnosis; in fact medical treatment which we present is effective for nerve dysfunction and may avoid surgery, but surgical exploration is mandatory when EMG is suggestive of a severe lesion, and localizes the specific site on anterior interosseous nerve entrapment.

On the use of 2,1,3-benzothiadiazole derivatives as selective live cell fluorescence imaging probes.

Newly designed 2,1,3-benzothiadiazole-containing fluorescent probes with four excited state intramolecular proton transfer (ESIPT) sites were successfully tested in live cell-imaging assays using a confluent monolayer of human stem-cells (tissue). All tested dyes were compared with the commercially available DAPI and gave far better results.

On the selective detection of duplex deoxyribonucleic acids by 2,1,3-benzothiadiazole fluorophores.

A series of 4,7-pi-extended 2,1,3-benzothiadiazoles (BTDs) with different molecular architectures, in particular, the organic dyes based on the 4-(arylethynyl)-7-(4-methoxy)-2,1,3-benzothiazole skeleton, can be used at very low concentrations (down to 10 microM) to detect DNA at 1 ppm in phosphate buffer solutions. Upon binding to DNA, these dyes showed an exponential increase in the fluorescence intensity (hyperchromic effect) and a red shift (1-5 nm) in the long-wavelength emission maxima. Pre-steady state kinetic experiments (stopped-flow) demonstrated the fast dye interaction with the biomacromolecules of DNA with an increase in fluorescence, especially with non-symmetrical BTDs containing an ethynyl spacer. An intercalation model could be proposed based on the photophysical properties, X-ray analysis, and theoretical calculations (ab initio). In this model, the intercalation occurs on the ethynyl side of the BTD, and as a consequence, the PhOMe portion is free to perform the ICT process with the BTD core and stabilizes it in the excited state.

Recent developments in the chemistry of deoxyribonucleic acid (DNA) intercalators: principles, design, synthesis, applications and trends.

In the present overview, we describe the bases of intercalation of small molecules (cationic and polar neutral compounds) in DNA. We briefly describe the importance of DNA structure and principles of intercalation. Selected syntheses, possibilities and applications are shown to exemplify the importance, drawbacks and challenges in this pertinent, new, and exciting research area. Additionally, some clinical applications (molecular processes, cancer therapy and others) and trends are described.

Steady-state kinetics of indole-3-glycerol phosphate synthase from Mycobacterium tuberculosis.

Indole-3-glycerol phosphate synthase (IGPS) catalyzes the irreversible ring closure of 1-(o-carboxyphenylamino)-1-deoxyribulose 5-phosphate (CdRP), through decarboxylation and dehydration steps, releasing indole-3-glycerol phosphate (IGP), the fourth step in the biosynthesis of tryptophan. This pathway is essential for Mycobacterium tuberculosis virulence. Here we describe the cloning, expression, purification, and kinetic characterization of IGPS from M. tuberculosis. To perform kinetic studies, CdRP was chemically synthesized, purified, and spectroscopically and spectrometrically characterized. CdRP fluorescence was pH-dependent, probably owing to excited-state intramolecular proton transfer. The activation energy was calculated, and solvent isotope effects and proton inventory studies were performed. pH-rate profiles were carried out to probe for acid/base catalysis, showing that a deprotonated residue is necessary for CdRP binding and conversion to IGP. A model to describe a steady-state kinetic sequence for MtIGPS-catalized chemical reaction is proposed.

Ionic liquid supported acid/base-catalyzed production of biodiesel.

The transesterification (alcoholysis) reaction was successfully applied to synthesize biodiesel from vegetable oils using imidazolium-based ionic liquids under multiphase acidic and basic conditions. Under basic conditions, the combination of the ionic liquid 1-n-butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)imide (BMINTf2), alcohols, and K2CO3 (40 mol %) results in the production of biodiesel from soybean oil in high yields (>98%) and purity. H2SO4 immobilized in BMINTf2 efficiently promotes the transesterification reaction of soybean oil and various primary and secondary alcohols. In this multiphase process the acid is almost completely retained in the ionic liquid phase, while the biodiesel forms a separate phase. The recovered ionic liquid containing the acid could be reused at least six times without any significant loss in the biodiesel yield or selectivity. In both catalytic processes (acid and base), the reactions proceed as typical multiphasic systems in which the formed biodiesel accumulates as the upper phase and the glycerol by-product is selectively captured by the alcohol-ionic liquid-acid/base phase. Classical ionic liquids such as 1-n-butyl-3-methylimidazolium tetrafluoroborate and hexafluorophosphate are not stable under these acidic or basic conditions and decompose.

New sensitive fluorophores for selective DNA detection.

4,7-Disubstituted benzothiadiazoles containing 1-arylethynyl and 4-methoxyphenyl groups are selective photoluminescent "light up" probes to duplex DNA with unprecedented sensibility in both spectrophotometric and spectrofluorimetric measurements.

Aspectos à tomografia de coerência óptica em retinosquise juvenil ligada ao cromossomo-X / Optical coherence tomography features in X-linked juvenile retinoschisis

Objetivos: Demonstrar a utilização e importância da tomografia de coerência óptica em dois casos de retinosquise ligada ao cromossomo X. Local: Departamento de Oftalmologia da Universidade de São Paulo, São Paulo (SP), Brasil. Método: Descrição dos achados oftalmoscópicos e tomografia de coerência óptica em dois casos de retinosquise ligada ao cromossomo X. Resultado: A tomografia de coerência óptica dos olhos com retinosquise ligada ao cromossomo X possibilitou a visibilização da esquise retiniana dentro da camada de fibras nervosas, confirmando a suspeita diagnóstica. Conclusão: A tomografia de coerência óptica é um método de imagem não invasivo, de não contato, e um poderoso método de exame para a detecção e monitorização de uma variedade de doenças maculares.