Search

Kinetic resolution of racemates with lipases is the preferred method for producing bioactive compounds. One strategy for obtaining highly enantioselective lipases that are stable in the organic media that are often used in these reactions is to improve existing lipases by protein engineering. In this work, we engineered the lipase LipC12, which has good stability in organic media, but only moderate enantioselectivity. Molecular docking with LipC12 identified V261 as a key position influencing, first, enantioselectivity in the transesterification of (RS)-1-phenylethanol and, second, activity in the hydrolysis of p-nitrophenyl octanoate. Variants were then obtained by site-directed mutagenesis, expressed in Escherichia coli, and their performance in these reactions was evaluated. Enzymes immobilized on Immobead 150 were used in the transesterification while free enzyme was used in the hydrolysis reaction. It was not possible to increase the hydrolytic activity and enantioselectivity simultaneously: some variants had increased enantioselectivity but lower hydrolytic activity, and others had increased hydrolytic activity but lower enantioselectivity. The best result for enantioselectivity was obtained for LipC12V261Q, with an increase of the E-value (for (R)-1-phenylethanol) from 46 to 110, however, its hydrolytic activity decreased 4-fold in comparison to LipC12wt. The highest hydrolytic activity was obtained for LipC12V261F, with a value almost 6-fold higher than that of LipC12wt. This variant also had an inverted enantiopreference (i.e. for (S)-1-phenylethanol), but with a very low E-value of only 4.

Moyamoya disease is a rare cause of stroke, radiologically characterised by progressive stenosis of the terminal portion of the internal carotid arteries and compensatory capillary collaterals. The discovery that RNF213, which encodes an unconventional E3 ubiquitin ligase, is the major susceptibility gene for moyamoya disease in people from east Asia has opened new avenues for investigation into the mechanisms of disease and potential treatment targets. The Arg4810Lys variant of the gene is most strongly associated with moyamoya disease, but the penetrance is lower than 1%, suggesting a synergistic relationship with additional environmental and genetic risk factors. White people carry less common non-Arg4810Lys variants of RNF213, which partly explains the lower prevalence of moyamoya disease in European countries and in the USA than in east Asian countries. Several monogenic moyamoya syndromes possess the radiological characteristics of moyamoya disease and have been associated with multiple genes and pathways involved in moyamoya angiopathy pathogenesis. Further clarification of the genetic and environmental factors that contribute to the emergence of moyamoya angiopathy could enable development of new treatment strategies for moyamoya disease.

Streptococcus mutans, a major cariogenic bacterium, expressing the collagen-binding protein Cnm induces cerebrovascular inflammation, resulting in the impairment of blood brain barrier integrity followed by cerebral bleeding. We here examined the association of Cnm-positive S. mutans with cerebral microbleeds (CMBs) in acute stroke patients selected from a single-center registry database. Of 428 patients who received oral bacterial examinations among 3154 stroke patients, 326 patients who harbored S. mutans were identified. After excluding four patients without imaging data, we compared 72 patients with Cnm-positive S. mutans and 250 with Cnm-negative S. mutans. Deep, lobar and infratentorial CMBs were observed in 46 (63.9%), 36 (50.0%), 25 (34.7%) patients with Cnm-positive S. mutans and 144 (57.6%), 114 (45.6%), 101 (40.4%) with Cnm-negative S. mutans. Possession of Cnm-positive S. mutans was related to higher numbers of both deep and lobar, but not infratentorial, CMBs (risk ratios 1.57 [1.07‒2.30], deep; 5.44 [2.50‒11.85], lobar). Statistical significance persisted after adjusting for age, sex, hypertension, stroke type, National Institutes of Health Stroke Scale score, and cerebral amyloid angiopathy (risk ratios 1.61 [1.14‒2.27], deep; 5.14 [2.78‒9.51], lobar). Our study indicated that reduction of Cnm-positive S. mutans may serve as a therapeutic approach for improving the prognosis of stroke patients.

Background: Cerebral amyloid angiopathy is a cerebrovascular disease directly implicated in Alzheimer’s disease pathogenesis through amyloid-β deposition. Growing evidence has shown a pivotal role of chronic neuroinflammation both in cerebral amyloid angiopathy and Alzheimer’s disease. Objective: The aim of this study was to investigate whether circulating levels of the complement 3, a crucial component of the innate immune system, are increased in patients with cerebral amyloid angiopathy. Methods: Serum complement 3 levels were retrospectively measured by a sandwich enzyme-linked immunosorbent assay in a single-center cohort of patients with mild cognitive impairment. The diagnosis of cerebral amyloid angiopathy was based on the modified Boston criteria. Logistic regression analysis was performed to identify the predictive factors for cerebral amyloid angiopathy. Results: We analyzed 55 mild cognitive impairment patients (mean age [standard deviation]: 76.3 [6.8] years; 33 [60% ] men). Complement 3 levels were significantly increased in cerebral amyloid angiopathy patients (n = 16) compared with those without cerebral amyloid angiopathy (n = 39) (median [interquartile range]: 0.43 [0.34–0.65] versus 0.35 [0.25–0.45], respectively; p = 0.040). Univariate and multivariate logistic regression analysis revealed that increased complement 3 levels were significantly associated with cerebral amyloid angiopathy. After selection of the best predictive model using stepwise selection, complement 3 was preserved as a significant independent predictive factor for cerebral amyloid angiopathy (odds ratio per 0.1 unit/mL increase [95% confidence interval]: 1.407 [1.042–1.899]; p = 0.026). Conclusion: Complement activation may play a pivotal role in cerebral amyloid angiopathy. Complement 3 may be a novel diagnostic biomarker for cerebral amyloid angiopathy.

Objective Treatment of mandibular lesions sometimes leads to the formation of large defects in the bone. Mandible reconstruction using grafted tissue, bone, and/or medical implants is required following the resection of such lesions. However, such grafts may not completely restore the continuity of the mandible. Facial transplantations using allografts are common, but mandibular allotransplantation has not been performed in humans, due to the unstable immune reaction of the complex tissue. In this study, mandibular transplantations performed under the effect of the immunosuppressive agent, FK506, were evaluated in dogs. Methods Mandibular allotransplantations were performed in 21 pairs of sex-blind beagle dogs. The left side of the mandible in the donor dog was transplanted to the left side of the mandible in the recipient dog. The FK506 doses were 0 mg for 3 dog pairs, 0.08 mg/kg/day for 3, 0.12 mg/kg/day for 5, 0.16 mg/kg/day for 5, and 0.32 mg/kg/day for 5 pairs. Twelve weeks after transplantation, the dogs were euthanized. Computed tomography (CT) and histological examinations of the transplanted sides were performed. Results Successful graft acceptance was dependent on immunosuppressive agent doses, and dogs administered more than 0.16 mg/kg/day FK506 showed 100 % survival. However, dogs administered 0.32 mg/kg/day FK506 showed significant weight loss 12 weeks after the surgery. CT findings showed thick mandibular cortical bone in the transplanted sites. Conclusions These results indicate that 0.16 mg/kg/day FK506 was the ideal dose for allotransplantations. Bone hyperplasia was observed in the transplanted sides, but it is not a disadvantage for allotransplantations.

AimsOcrelizumab is a humanized monoclonal antibody that selectively targets CD20-positive B cells and is indicated for treatment of patients with relapsing forms of multiple sclerosis (RMS) or primary progressive multiple sclerosis (PPMS). The pharmacokinetics and pharmacodynamics of ocrelizumab in patients with RMS or PPMS were assessed.MethodsA population pharmacokinetic model was developed based on data from the Phase II study and the Phase III studies OPERA I and OPERA II in patients with RMS. Data from the ORATORIO Phase III study in patients with PPMS became available after model finalization and was used for external model evaluation.ResultsThe ocrelizumab serum concentration vs time course was accurately described by a 2-compartment model with time-dependent clearance. Body weight was found to be the main covariate. The area under the concentration-time curve over the dosing interval was estimated to be 26% higher for patients with RMS weighing 90kg when compared with the 60-90kg group. The terminal half-life of ocrelizumab was estimated as 26days. The extent of B-cell depletion in blood, as the pharmacodynamic marker, was greater with increasing ocrelizumab exposure.ConclusionThe pharmacokinetics of ocrelizumab was described with pharmacokinetic parameters typical for an immunoglobulin G1 monoclonal antibody, with body weight as the main covariate. The pharmacokinetics and B-cell depletion in blood were comparable across the RMS and PPMS trials, and the extent of blood B-cell depletion was greater with higher exposure.

To better understand the cancer risk posed by radiation and the development of radiation therapy resistant cancer cells, we investigated the involvement of the cancer risk factor, APOBEC3B, in the generation of radiation-induced mutations. Expression of APOBEC3B in response to irradiation was determined in three human cancer cell lines by real-time quantitative PCR. Using the hypoxanthine-guanine phosphoribosyl transferase (HPRT) mutation assay, mutations in the HPRT gene caused by irradiation were compared between APOBEC3B-deficient human hepatocellular carcinoma (HepG2) cells [APOBEC3B knocked out (KO) using CRISPR-Cas9 genome editing] and the parent cell line. Then, HPRT-mutated cells were individually cultured to perform PCR and DNA sequencing of HPRT exons. X-Irradiation induced APOBEC3B expression in HepG2, human cervical cancer epithelial carcinoma (HeLa) and human oral squamous cell carcinoma (SAS) cells. Forced expression of APOBEC3B increased spontaneous mutations. By contrast, APOBEC3B KO not only decreased the spontaneous mutation rate, but also strongly suppressed the increase in mutation frequency after irradiation in the parent cell line. Although forced expression of APOBEC3B in the nucleus caused DNA damage, higher levels of APOBEC3B tended to reduce APOBEC3B-induced γ-H2AX foci formation (a measure of DNA damage repair). Further, the number of γ-H2AX foci in cells stably expressing APOBEC3B was not much higher than that in controls before and after irradiation, suggesting that a DNA repair pathway may be activated. This study demonstrates that irradiation induces sustained expression of APOBEC3B in HepG2, HeLa and SAS cells, and that APOBEC3B enhances radiation-induced partial deletions.

Patients with myelodysplastic syndromes (MDS) with progenitors expressing CD41 (CD41+ MDS) showed a poor prognosis in a previous study but their detailed characteristics remain unclear.One hundred thirty-seven subjects at our institution were diagnosed with excess blasts (EB)-1, EB-2, and acute myeloid leukemia with a low blast count (20%-30%). The immunophenotypes of progenitor cells in their bone marrow (BM) were determined by CD45-gating flow cytometry. A false-positive reaction to CD41 was eliminated by examining the flow cytometry data of lymphocytes and monocytes in addition to progenitors and by examining CD42b in histological sections. The characteristics were compared between CD41+ and CD41- MDS patients.Forty-three patients (31%) were CD41+. Additionally, 91% of the CD41+ MDS patients were very high-risk defined by the Revised International Prognostic Score System, which was higher than in patients with CD41- MDS (p = 0.015). Approximately 60% of the CD41+ MDS patients had a monosomal karyotype and very poor cytogenetics, which was higher than in CD41- MDS patients (p 0.001). Normal cytogenetics was less common in CD41+ patients (p = 0.0016). Blasts with bleb formation were more abundant in CD41+ MDS patients (p = 0.026). All CD41+ MDS patients were positive for CD13 and were mostly positive for CD33. The frequency of aberrant expression of other antigens on progenitors was similar between CD41+ and CD41- MDS patients.We determined clinical, immunophenotypic, and cytogenetic characteristics of CD41+ MDS patients. Further studies are needed to improve the survival of these patients.

The role of commensal microbiota in systemic diseases, including brain diseases, has attracted increasing attention. Oral infectious diseases, such as dental caries and periodontitis, are also involved in cerebrovascular diseases and cognitive impairment. Cerebral microbleeds (CMBs) and intracerebral hemorrhage due to small vessel disease (SVD), are presumably associated with a high risk of vascular cognitive impairment and stroke. We previously reported thatFifteen domestic institutes will be enlisted to enroll 230 patients who have at least one CMB in the deep brain area and develop a stroke within the past year. The prevalence of Cnm-positiveThe 2-year longitudinal prospective cohort study is expected to establish the role of Cnm-positive

Made available in DSpace on 2022-05-01T05:29:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-09-01 Background: Antimicrobial photodynamic therapy (aPDT) is used as an adjunct to endodontic treatment to enhance microbial reduction in the root canal system. However, studies evaluating the impact of laser ablation with Indocyanine Green (ICG) are scarce. Thus, this in vitro study aimed to evaluate the efficiency of laser ablation with ICG using different laser parameters compared with aPDT using photosensitizer methylene blue (MB) and curcumin (CUR) on the reduction of E. faecalis biofilms on root canals. Methods: Forty-nine human premolars were used after biomechanical instrumentation for standardization. The root canals were contaminated with E. faecalis for 10 days to form biofilms, and divided into 7 groups (n = 7): 0.01% MB activated by red laser - MB+RL; 0.05% CUR activated by blue LED – CUR+BL; 0.05% ICG activated by infrared diode laser (2.5 W power, 30 ms interval, and 30 ms duration) - ICG+DL 2.5/30/30; 0.05% ICG activated by infrared diode laser (2.5 W power, 300 ms interval, and 100 ms duration) - ICG+DL 2.5/300/100; 0.05% ICG activated by infrared diode laser (3 W power, 300 ms interval, and 100 ms duration) - ICG+DL 3/300/100; sterile saline solution (negative control) - NC; and 2.5% NaOCl (positive control) - PC. Root canal sampling was performed prior to and immediately after the different treatment protocols. Data were submitted to One- and Two-Way ANOVA, followed by Student-Newman-Keuls test or Fisher LSD's test (α = 0.05). Results: All aPDT protocols promoted significant CFU reductions compared with the NC; the highest CFU reduction was observed for PC (p < 0.05). Among the protocols the highest CFU reduction was promoted by laser ablation with ICG+DL 3/300/100 (p < 0.05) except compared with aPDT using CUR+BL (p > 0.05). Conclusions: Laser ablation protocol using ICG+DL proved to be efficient in reducing E. faecalis biofilms, especially when activated at 3/300/100 configuration. Department of Preventive and Restorative Dentistry São Paulo State University (UNESP) School of Dentistry Department of Restorative Dentistry São Paulo State University (UNESP) School of Dentistry Department of Preventive and Restorative Dentistry São Paulo State University (UNESP) School of Dentistry Department of Restorative Dentistry São Paulo State University (UNESP) School of Dentistry

Evidence-based analgosedation in severe pediatric traumatic brain injury (pTBI) management is lacking, and improved pharmacological understanding is needed. This starts with increased knowledge of factors controlling the pharmacokinetics (PK) of unbound drug at the target site (brain) and related drug effect(s). This prospective, descriptive study tested a pediatric physiology-based pharmacokinetic software model by comparing actual plasma and brain extracellular fluid (brainECF) morphine concentrations with predicted concentration-time profiles in severe pTBI patients (Glasgow Coma Scale [GCS], ≤8). Plasma and brainECF samples were obtained after legal guardian written consent and were collected from 8 pTBI patients (75% male; median age, 96 months [34.0–155.5]; median weight, 24 kg [14.5–55.0]) with a need for intracranial pressure monitoring (GCS, ≤8) and receiving continuous morphine infusion (10–40 μg/kg/h). BrainECF samples were obtained by microdialysis. BrainECF samples were taken from “injured” and “uninjured” regions as determined by microdialysis catheter location on computed head tomography. A previously developed physiology-based software model to predict morphine concentrations in the brain was adapted to children using pediatric physiological properties. The model predicted plasma morphine concentrations well for individual patients (97% of data points within the 90% prediction interval). In addition, predicted brainECF concentration-time profiles fell within a 90% prediction interval of microdialysis brainECF drug concentrations when sampled from an uninjured area. Prediction was less accurate in injured areas. This approach of translational physiology-based PK modeling allows prediction of morphine concentration-time profiles in uninjured brain of individual patients and opens promising avenues towards evidence-based pharmacotherapies in pTBI.

Superconductivity in FeSe was investigated under high pressure through the measurements of DC magnetization by using a diamond anvil cell. We successfully observed that the disappearance of the superconductivity as a results of the appearance of the non-superconducting ortho I\hspace{-1pt}I phase above $\sim$7 GPa ($\sim$5 GPa), when Ar (glycerin) is used as the pressure transmitting media. Contrary to this, it has been found that the superconductivity even survives under pressure above 7 GPa, when the thickness ($t$) of a platelet-single crystal specimen is reduced. The survival of the superconductivity above 7 GPa is consistent with a previous observation under hydrostatic pressure by using a cubic anvil apparatus, suggesting that the hydrostaticity of the pressure is improved by reducing $t$. It is also inferred that the appearance of the ortho I\hspace{-1pt}I phase is due to uniaxial stress along the [001] direction., 6 pages, 4 figures

Supplemental Digital Content is available in the text., Background and Purpose: Cerebral microbleeds (CMB) are associated with stroke and cognitive impairment. We previously reported a high prevalence of CMB in people with Streptococcus mutans expressing Cnm, a collagen-binding protein in the oral cavity. S. mutans is a major pathogen responsible for dental caries. Repeated challenge with S. mutans harboring the cnm gene encoding Cnm induced cerebral bleeding in stroke-prone spontaneously hypertensive rats. The purpose of this longitudinal study is to examine the relationship of cnm-positive S. mutans to the development of CMB. Methods: We retrospectively investigated patients with stroke receiving oral microbiological examination and head 3T magnetic resonance imaging evaluations twice in the period 2014 to 2019, allowing >180-day interval. Patients with cnm-positive S. mutans were compared with those without. Quasi-Poisson regression models were used to explore associations between cnm-positive S. mutans and the increase in number of CMB between the 2 magnetic resonance imaging scans. Results: A total of 111 patients were identified; 21 (19%) with cnm-positive S. mutans and 90 (81%) without. Clinical history, including blood pressure and the use of antithrombotic agents, were comparable between the 2 groups. New CMB were more commonly observed in patients with cnm-positive S. mutans (52% versus 23%; P=0.008). The incidence of CMB was significantly higher in the group with cnm-positive S. mutans, especially in deep areas, (incidence rate ratios [95% CI], 5.1 [1.9–13.6] for CMB in any brain region; 15.0 [5.4–42.0] for deep CMB), which persisted after adjusting for age, sex, hypertension, and renal impairment (4.7 [1.8–11.9] for CMB in any brain region; 13.9 [4.3–44.5] for deep CMB). Conclusions: This study demonstrates that cnm-positive S. mutans is associated with an increased incidence of CMB. Treatment for cnm-positive S. mutans infection may be a novel microbiota-based therapeutic approach for stroke and cognitive impairment.

O objetivo deste artigo é fazer uma revisão de literatura que engloba os achados neurológicos do SARS-CoV-2, principalmente relacionados com a perda de memória, para auxiliar no maior entendimento da doença, e nos possíveis aspectos fisiopatológicos que a originam. Foram pesquisados artigos publicados de 2020 a 2021, através da base de dados PubMed, CAPES, UpToDate e Scielo, usando as palavras chave ‘Covid-19’, ‘Memory’, ‘Brain Fog’, ‘Long-Hauler’, ‘SARS-CoV-2’, ‘Nervous System’, ‘Cognitive Deficit’, ‘Coronavirus’, Memory-Loss’, ‘Alzheimer’, ‘Neuro-Covid’, ‘Neuro Invasion’. Entre os relatos de perda de memória associados ao COVID-19, a fisiopatologia mais comum encontrada foi a alteração indireta por meio de mecanismos inflamatórios no SNC, seguida de alterações diretas no hipocampo e regiões corticais. Houve também a descrição de “brain frog”, alterações cognitivas, cefaleia, distúrbios do sono, alteração de humor, depressão e ansiedade. As mudanças descritas atingiram principalmente os pacientes a longo prazo após o processo de recuperação da doença, mas também foram relatadas alterações agudas. Os achados neurológicos da COVID-19 são semelhantes aos encontrados em outras doenças de etiologia viral. Existe ainda a possibilidade de agravamento de condições clínicas pré existentes como parkinsonismos e alzheimer, além de comorbidades como diabetes, hipertensão e doença renal possibilitarem a piora do quadro clínico dos pacientes. Devemos também nos atentar a possibilidade da manifestação neural inicial da doença ser “brain-frog”. Dessa forma, a correlação entre perda de memória e SARS-CoV-2 demanda mais estudos e é importante para a pesquisa científica sobre os achados neurológicos da pandemia atual.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common form of hereditary cerebral small vessel disease. Previous neuroimaging studies have suggested loss of hippocampal volume is a pathway for cognitive impairment in CADASIL. We used unbiased stereological methods to estimate SMI32-positive and total numbers and volumes of neurons in the hippocampal formation of 12 patients with CADASIL and similar age controls (young controls) and older controls. We found densities of SMI32-positive neurons in the entorhinal cortex, layer V, and cornu ammonis CA2 regions were reduced by 26%–50% in patients with CADASIL compared with young controls (p < 0.01), with a decreasing trend observed in older controls in the order of young controls> older controls ≥ CADASIL. These changes were not explained by any hippocampal infarct or vascular pathology or glial changes. Our results suggest notable loss of subsets of projection neurons within the hippocampal formation that may contribute to certain memory deficits in CADASIL, which is purely a vascular disease. It is likely that the severe arteriopathy leads to white matter damage which disconnects cortico-cortical and subcortical-cortical networks including the hippocampal formation., Highlights • Hippocampal volume loss was associated with cognitive dysfunction in CADASIL. • SMI32+ neurons in entorhinal cortex (V) and CA2 regions were reduced in CADASIL. • Hippocampal cellular changes were not explained by any infarct pathology. • Neuronal volumes or glial cell numbers per neuron were not changed. • Selective loss of projection neurons within the hippocampal formation in CADASIL.

Objectives Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is characterised by strategic white matter (WM) hyperintensities on MRI. Pathological features include WM degeneration, arteriolosclerosis, lacunar infarcts and the deposition of granular osmiophilic material. Based on the hypothesis that the gliovascular unit is compromised, we assessed the nature of astrocyte damage in the deep WM of CADASIL subjects. Methods We evaluated post‐mortem brains from CADASIL, cerebral small vessel disease, similar age cognitively normal and older control subjects. Standard immunohistochemical, immunofluorescent and unbiased stereological methods were used to evaluate the distribution of astrocytes, microvessels and autophagy markers in five different brain regions. Results Compared to the controls, the deep WM of CADASIL subjects overall showed increased numbers of glial fibrillary acidic protein (GFAP)‐positive clasmatodendritic astrocytes (P=0.037) and a decrease in the percentage of normal appearing astrocytes (P=0.025). In accord with confluent WM hyperintensities , the anterior temporal pole contained abundant clasmatodendritic astrocytes with displaced aquaporin 4 immunoreactivity. Remarkably, we also found strong evidence for the immunolocalisation of autophagy markers including microtubule associated protein 1, light chain 3 (LC3) and sequestosome 1/p62 and Caspase‐3 in GFAP‐positive clasmatodendritic cells, particularly within perivascular regions of the deep WM. LC3 was co‐localised in more than 90% of the GFAP‐positive clasmatodendrocytes. Conclusions Our novel findings show astrocytes undergo autophagy‐like cell death in CADASIL, with the anterior temporal pole being highly vulnerable. We propose astrocytes transform from normal appearing type A to hypertrophic type B and eventually to clasmatodendritic type C cells. These observations also suggest the gliovascular unit of the deep WM is severely impaired in CADASIL.

Genome-wide association and candidate gene studies have identified COL4A1 and COL4A2 as risk genes for intracerebral hemorrhage (ICH), suggesting that the disrupted collagen architecture could be a contributory factor in disease onset. Environmental factors that disrupt the vascular collagen architecture may therefore bring about gene–environmental interactions. Certain oral strains of Streptococcus mutans expressing Cnm, a collagen-binding protein, have been found to be responsible for ICH in a preclinical study. In support of this finding, a population-based study showed a close association between Cnm-positive Streptococcus mutans with cerebral microbleeds, a precursor of ICH, and a hospital-based study between such bacteria with cerebral microbleeds and ICH. Taken together, these findings suggest that Cnm-positive Streptococcus mutans serve as an important environmental factor in ICH.

Diagnosis of myelodysplastic syndromes (MDS) is not straightforward when objective data, such as blast excess and abnormal cytogenetics, are lacking. Expert laboratories use flow cytometry (FCM) to help diagnose MDS. However, most of FCM protocols for MDS are complex, requiring a high level of expertise and high cost. We have reported a FCM mini-panel consisting of four FCM parameters (so-called Ogata score), which is simple to conduct and inexpensive. In this paper, to refine this mini-panel, we have introduced a new FCM parameter, which quantifies CD33 expression on CD34+ cells (called Granulocyte/CD34 cell CD33 ratio). Bone marrow cells from MDS without blast excess (low-grade MDS) and controls were stained with CD34, CD45, and CD33 and analyzed for five parameters ("Granulocyte/CD34 cell CD33 ratio" plus four parameters in the Ogata score). By a multivariate logistic regression model, only three parameters, including "Granulocyte/CD34 cell CD33 ratio" had statistically significant power for diagnosing low-grade MDS. Based on the results, we constructed a new scoring system, which showed approximately 50% sensitivity and more than 95% specificity in diagnosing low-grade MDS. Our revised mini-panel is suitable for screening samples suspected for MDS and provides a basis for further improvement in diagnostic FCM protocols for MDS.

Excessive intake of a high-calorie diet has been implicated in the development of non-alcoholic fatty liver disease (NAFLD). Several studies have investigated the effect of NAFLD on liver regeneration, but the effects of simple steatosis have been found to be inconsistent. We aimed to assess whether the initial phase of diet-induced lipid accumulation, induced by a diet containing moderate levels of fat, impairs liver regeneration after partial hepatectomy (PHx) of mice.Male ddY mice are prone to obesity, even when fed a relatively low-fat diet (FD). A model of early simple steatosis was created by feeding a 10% FD for 6 weeks to male ddY mice. Liver regeneration rate, DNA synthesis in hepatocytes, and DNA damage were then assessed.FD-fed mice had a slightly higher body mass (44.5 ± 2.6 grams vs. 48.1 ± 3.6 grams, P 0.05), but not liver mass (2.55 ± 0.37 grams vs. 2.69 ± 0.26 grams). Lipid droplets appeared in FD-fed mouse hepatocytes and Oil Red O staining was five times as intense as in control mice. In FD-fed mice, liver regeneration rate after two-thirds PHx was lower and impaired DNA replication was also observed. FD induced the expression of the cyclin-dependent kinase inhibitor p27KIP, but not p21CIP. Moreover, greater histone 2A.X phosphorylation was observed, indicating that FD caused DNA damage.Even short-term feeding of a moderately high FD to male ddY mice results in lipid accumulation in hepatocytes, DNA damage, and greater expression of p27KIP, implying lower DNA synthesis.

The objective of this paper is to describe the experiential marketing through sports sponsorship strategy used by a sporting goods trade company that act in a business-to-business relationship (B2B) context. This paper is based on authors who argue that in the relationships of the B2B segment, can be influenced by emotional attributes and the evidence that sports sponsorship is a way to communicate these attributes. From the Content Analysis of semistructured interviews, four categories were identified: 1) Organizational Culture of the Company; 2) Context and History of Sponsorship actions, 3) Sports Sponsorship; 4) Returns with Experiential Marketing, where the considerations point out that Experiential Marketing through a sport sponsorship in the B2B context must count on professionalism, consistency between the corporate culture and sponsored object, as well as establishing action plans so that the desired return is won.