bluuebunny/crossref_metadata_2025 huggingface dataset:
Assessing the implementation of food defense requirements in industrial meat-based food processors
Authors: Praia Elisabete Filipa , Henriques Ana Rita
Published: April 2021

Abstract Food defense is the effort to protect food from causing harm to the consumer, including security procedures to mitigate intentional acts of adulteration. To assure entry in new markets, food companies need to develop and implement food defense strategies through third-party certification. Although there are some programs designed to assist food business operators in creating effective food defense strategies, this is still not regarded as a priority by food companies. As a first aim of this work, a first-party audit of two meat-producing industries was performed to verify the implementation of food defense requirements. The second purpose of the work was to compare vulnerabilities identified in those two food industries with the ones detected in other previously certified food units in Portugal. For such, a food defense requirements checklist was prepared for the audit and a private certification database was consulted to compare audit results with those from other food business operators certified by at least one international food defense standard. Audit results revealed that both industries were above 50% in overall compliance regarding food defense requirements; still the main vulnerabilities were related to the lack of a food defense plan, the failure to identify critical areas, ineffective warning systems and no training in food defense. Similar vulnerabilities were detected in other certified national food business operators, leading to the conclusion that implementation of food defense requirements seems to be underestimated. As an intervention strategy proposal, food defense training would be of upmost importance to get staff and managers acquainted with the concept.
Effect of microencapsulation and mango peel powder on probiotics survival in ice cream
Authors: Hayayumi-Valdivia María , Márquez-Villacorta Luis Francisco , Pretell-Vásquez Carla Consuelo
Published: April 2021

Abstract This study evaluated the effect of microencapsulation and addition of mango peel powder on the survival of Lactobacillus acidophilus and Bifidobacterium lactis, overrun, apparent viscosity, and overall acceptability of symbiotic ice cream during storage at -20 °C for 180 days. Six formulations of vanilla-flavored ice cream were prepared: three with addition of probiotic cultures at a concentration of 108 CFU/g and 0, 2%, and 3% mango peel powder microencapsulated in a sodium alginate matrix, and three with free addition. Analytical evaluations were performed after 1, 30, 60, 90, 120 and 180 days of storage. The results showed that microencapsulation of probiotics and prebiotics statistically influenced (p < 0.05) the characteristics evaluated. The formulation with microencapsulated probiotics and 2% mango peel powder was considered as the best product. This formulation is promising for future commercial application as a functional food because, at the end 180 days of storage, it showed probiotics population >106 CFU/g, 72.97% overrun, 292 mPA apparent viscosity, and good overall acceptance (7.6 points) equivalent to “I like it very much”.
Texture profile of surimi-like material from mechanically deboned turkey meat
Authors: Massingue Armando Abel , Paula Mariele Maria de Oliveira , Rocha Ana Paula , Haddad Gabriela de Barros Silva , Carmo Eloá Lourenço do , Ramos Alcinéia de Lemos Souza , Ramos Eduardo Mendes
Published: April 2021

Abstract The study aimed to evaluate the textural properties of turkey surimi-like (TSL) gels from mechanically deboned turkey meat (MDTM) by washing procedures. The effects of water, sodium chloride, sodium bicarbonate, and potassium phosphate buffer washing solutions, cycles, and salt addition were investigated. Sodium bicarbonate washing solution produced lightened and whitened cooked gels. Addition of salt reduced (p < 0.05) cooking loss and increased (p < 0.05) hardness and chewiness. TSL gels from sodium bicarbonate had the lowest (p < 0.05) cooking loss and hardness, and increased (p < 0.05) adhesiveness after two washing cycles. Scanning electron micrographs showed fine-stranded gel structures formed at low ionic strength and coarsely aggregated gel structures at high ionic strength. Salt addition improved the TSL gelling properties. Two washing cycles of MDTM with sodium bicarbonate solution generate high-quality colorless gels of TSL with proper textural properties and could be suggested as an alternative meat source in the manufacturing of surimi-based foodstuffs.
Espiridião, o carnaval e uma estética do moderno na Florianópolis de Salim Miguel
Authors: Kremer Natan Schmitz , Vaz Alexandre Fernandez
Published: April 2021

Resumo Neste artigo, analisamos algumas chaves provocadas pelo conto “Carnaval: casos de Espiridião”, narrativa que compõe o livro de estreia de Salim Miguel, Velhice e outros contos, para construir um quadro que se ocupa tanto da imagem de Florianópolis que nele está amalgamada quanto da própria experiência modernista na qual a obra se insere. Assim, discutimos alguns aspectos da cidade que se moderniza e em como esses elementos aparecem, primeiro, em um plano discursivo, o que aponta para uma série de contradições que nos faz atentar à forma. Nessa análise, apontamos para como o conto mimetiza a estrutura dual das elites da cidade e, ao mesmo tempo, impõe, pela montagem, a presença do modernismo como possibilidade de diferenciação.
O livro como forma de arte: a contribuição de Julio Plaza na produção e teorização do livro de artista no Brasil
Authors: Mattar Marina Ribeiro
Published: April 2021

Resumo Este artigo busca reunir e analisar a contribuição de Julio Plaza na construção de duas importantes questões para o entendimento da produção editorial: a noção de “independente” voltada à produção e circulação de livros e o campo teórico e conceitual do livro de artista, visto como produto de certa prática independente de criação. Para tanto, investigamos a produção teórica de Plaza voltada a essa temática, veiculada em revistas e jornais nos anos 1980, assim como a literatura crítica que investiga o campo editorial brasileiro nos dias atuais. Fez-se relevante também abordar os conceitos do livro de artista na tentativa de compreender como o termo vem sendo desenhado em suas linhas teóricas. Notou-se que os livros de artista se mostraram como produtos vinculados a modos independentes de produção, sendo, geralmente, frutos da arte e poesia de vanguarda, onde estava inserido Plaza. Desta forma, compreendeu-se, neste resgate histórico da atuação do artista e teórico no campo editorial brasileiro, que a autopublicação e autoedição foi - e continua sendo - uma maneira de “furar” a bolha da dita “edição sistema”.
Efficacy of speech therapy in post-intubation patients with oropharyngeal dysphagia: a randomized controlled trial
Authors: Turra Giovana Sasso , Schwartz Ida Vanessa Doederlein , Almeida Sheila Tamanini de , Martinez Chenia Caldeira , Bridi Maristela , Barreto Sérgio Saldanha Menna
Published: April 2021

ABSTRACT Purpose to verify the efficacy of speech therapy in the early return of oral intake in patients with post-orotracheal intubation dysphagia. Methods It was a double-blinded randomized controlled trial for two years with patients of intensive care units of a hospital. Study inclusion criteria were orotracheal intubation>48hours, age≥18 years old, clinical stability, and dysphagia. Exclusion criteria were tracheotomy, score 4 to 7 in the Functional Oral Intake Scale (FOIS), neurological disorders. Patients were randomized into speech treatment or control group (ten days of follow-up). The treated group (TG) received guidance, therapeutic techniques, airway protection and maneuvers, orofacial myofunctional and vocal exercises, diet introduction; the control group (CG) received SHAM treatment. Primary outcomes were oral intake progression, dysphagia severity, and tube feeding permanence. Results In the initial period of study, 240 patients were assessed and 40 (16.6%) had dysphagia. Of this, 32 patients met the inclusion criteria, and 17 (53%) received speech therapy. Tube feeding permanence was shorter in TG (median of 3 days) compared to CG (median of 10 days) (p=0.004). The size effect of the intervention on tube feeding permanence was statistically significant between groups (Cohen's d=1.21). TG showed progress on FOIS scores compared to CG (p=0.005). TG also had a progression in severity levels of Dysphagia protocol (from moderate to mild dysphagia) (p<0.001). Conclusion Speech therapy favors an early progression of oral intake in post-intubation patients with dysphagia. Clinical Trial Registration: RBR-9829jk.
Terapia ocupacional e educação: as proposições de terapeutas ocupacionais na e para a escola no Brasil1
Authors: Pereira Beatriz Prado , Borba Patrícia Leme de Oliveira , Lopes Roseli Esquerdo
Published: April 2021

Resumo Terapeutas ocupacionais têm contribuído e ambicionam dar continuidade à criação de soluções para problemáticas no âmbito do setor da Educação. Assim, o objetivo da pesquisa da qual decorre este artigo foi conhecer e analisar as proposições que terapeutas ocupacionais, no Brasil, vêm fazendo para e na Escola. Para tanto, lançou-se mão de uma revisão de mapeamento na biblioteca SciELO e nos periódicos Cadernos Brasileiros de Terapia Ocupacional e Revista de Terapia Ocupacional da Universidade de São Paulo, em período aberto até o fim de 2018. Com esse levantamento e sua sistematização, foram reunidos 36 textos que articulavam “terapia ocupacional e escola/educação”. Paralelamente, durante 2017, foram realizadas seis entrevistas com pesquisadoras brasileiras que atuavam na interface terapia ocupacional e educação. O conjunto de dados reunidos e sua análise levaram à elaboração de quatro eixos temáticos que configuraram e delimitaram a argumentação utilizada para sustentar e problematizar proposições terapêutico-ocupacionais em diálogo com o setor da Educação, especificamente nas escolas. Ficou evidente uma prevalência de práticas voltadas às crianças com deficiência, em uma perspectiva individualizada e relacionada à produção da/na saúde. Paripassu, há o crescimento de proposições voltadas a crianças em geral na educação infantil e a jovens em situação de vulnerabilidade social. Pontua-se que o estofo para designar a educação como um campo particular de intervenção do terapeuta ocupacional, com um recorte metodológico próprio, um saber técnico e populações específicas, ainda está em construção e em disputa no interior da própria terapia ocupacional.
Sensitization with Eustrongylides sp. (Nematoda: Dioctophymatidae) antigens induce production of specific IgG and IgE in murine model
Authors: Kuraiem Bianca Porto , Verícimo Maurício Afonso , Knoff Marcelo , Mattos Danuza Pinheiro Bastos Garcia de , São Clemente Sérgio Carmona de
Published: April 2021

Abstract Fish consumption plays an important role in human diet. Hoplias malabaricus, commonly known as traíra, is a freshwater fish widely appreciated in several Brazilian states and frequently infected by Eustrongylides sp. fourth-instar larvae (L4). The aim of the present study was to evaluate allergenic potential of Eustrongylides sp. L4 crude extract (CEE). BALB/c mice were immunized intraperitoneally (IP) by 10 μg CEE with 2 mg of aluminum hydroxide on days 0 and 35. Specific IgG and IgE antibody levels were determined after immunization and cellular immunity was evaluated by assessing intradermal reaction in ear pavilion. Epicutaneous sensitization was performed in dorsal region by antigen exposure using a Finn-type chamber containing 50 μg of CEE or saline solution, followed by evaluation of specific antibody levels. IP immunization resulted in a gradual increase in IgG antibody levels and transitory IgE production. Significant increase in ear thickness was observed in cellular hypersensitivity reaction. In case of antigen exposure by epicutaneous route, CEE was able to induce meaningfully increased levels of specific IgG and IgE antibodies as well as heightened cellular immunity. Both intraperitoneal immunization and epicutaneous contact with Eustrongylides sp. larval antigens were observed for first time to be capable of inducing immunological sensitization in mice.
Upper Paraná River floodplain: synthesizing knowledge over time
Authors: Osório Nicolli Cristina , Rodrigues Liliana
Published: April 2021

Abstract: Aim Considering the great environmental and social impact that floodplains have, several studies have been performed in the upper Paraná River floodplain. In this study, we applied a scientometric analysis to understand the evolution of scientific knowledge generated in the Upper Paraná River floodplain, based on studies published in this area. Methods We searched the literature published from 1995 to 2018 using the indexed database - Thomson ISI Web of Science. The following information was obtained for each scientific article: a) year of publication; b) place of research; c) research institution; d) study approach; e) environments; f) subject matter; g) theoretical approach; h) taxonomic groups. Results We found 435 published articles from different study areas (such as: exact and earth sciences, environmental sciences, social and human sciences, health sciences) that used this floodplain as a study area. The linear regression model showed an increase in the number of articles throughout the years. Studies with ecological themes were most prominent, ensuring research with various theoretical approaches and different biological communities used as proxies. Conclusions With this study, we conclude that long-term research was fundamental in generating data to address ecological issues and describe the dynamics of this plain. Studies such as these must have more political and financial support to develop basic scientific studies that can subsidize data for conservation programs and government policies, in order to maintain ecosystem services such as fishing and the availability of quality water.
Lack of In Vivo Function of Osteopontin in Experimental Anti-GBM Nephritis
Authors: BONVINI JOHN MICHAEL , SCHATZMANN URSULA , BECK-SCHIMMER BEATRICE , SUN LI KANG , RITTLING SUSAN R. , DENHARDT DAVID T. , LE HIR MICHEL , WÜTHRICH RUDOLF P.
Published: April 2021

Osteopontin (Opn) is a potent chemoattractant for mononuclear cells that is upregulated in various inflammatory states of the kidney. Opn is believed to contribute to mononuclear cell infiltration and renal injury. The importance of Opn was examined in vivo in rapidly progressive glomerulonephritis in Opn knockout mice. Glomerulonephritis was induced by intravenous injection of rabbit anti-mouse glomerular basement membrane antiserum in mice that had been presensitized to rabbit IgG. Immunologic responsiveness to rabbit IgG (assessed by cutaneous delayed-type hypersensitivity and antibody titers) showed no significant difference between wild-type and Opn -/- mice. Proteinuria was also similar in both groups. Glomerular crescent formation was not different in Opn +/+ and -/- groups (26 ± 6% versus 29 ± 7%). Tubulointerstitial infiltration was assessed qualitatively and showed no significant difference between the two genotypes. Formation of thrombi in the glomerular capillaries on a scale from 0 to 3 also showed no significant difference (1.3 ± 0.3 for Opn +/+ and 1.4 ± 0.3 for Opn -/- mice). Northern blot analysis of total kidney RNA showed a 5.1-fold increase of Opn expression in Opn +/+ mice compared with untreated controls and the absence of expression in Opn -/- mice, as expected. Regulated upon activation, normal T cell expressed and secreted (RANTES) and monocyte chemoattractant protein-1 mRNA levels were also markedly upregulated with no significant difference between the two strains, excluding compensatory overexpression of these two chemokines in Opn -/- mice. It is concluded that the known upregulation of Opn in murine anti-glomerular basement membrane nephritis does not significantly contribute to the glomerular and tubulointerstitial mononuclear cell infiltration in this model.
A Locus for Adolescent and Adult Onset Familial Focal Segmental Glomerulosclerosis on Chromosome 1q25-31
Authors: TSUKAGUCHI HIROYASU , YAGER HENRY , DAWBORN JOHN , JOST LUIS , COHLMIA JERRY , ABREU PATRICIA F. , PEREIRA APARECIDO B. , POLLAK MARTIN R.
Published: April 2021

Focal segmental glomerulosclerosis is a nonspecific renal lesion observed both as a primary (idiopathic) entity and in a secondary form, typically in association with reduced functional renal mass. Familial forms have been observed and two loci for autosomal dominant FSGS have been mapped. This study shows that an adolescent/adult form of recessive FSGS maps to a locus on chromosome 1q25-31, which overlaps with a region previously identified as harboring a locus for an early childhood onset recessive form of nephrotic syndrome (SRN1). Evaluation of a large family demonstrated linkage with a maximum two-point lod score of 3.98 at D1S254 and D1S222. Lod score calculations support the conclusion of linkage in four of five additional families. Haplotype analysis suggests that this FSGS gene is located in a 19-cM region flanked by D1S416 and D1S413, of which 6 cM overlaps with SRN1, suggesting that these distinct clinical subsets of kidney disease may be allelic. These regions may also overlap with the syntenic region of the glomerulosclerosis susceptibility locus in the BUF/Mna rat. Because the presentation of FSGS may be subtle, inherited FSGS may be much more common than generally realized and grossly underestimated because of the absence of clear familial patterns. This result increases the suspicion that polymorphisms at this locus may contribute to sporadic FSGS.
Induction of CD14 in Tubular Epithelial Cells During Kidney Disease
Authors: MORRISSEY JEREMIAH , GUO GUANGJIE , MCCRACKEN RUTH , TOLLEY TIMOTHY , KLAHR SAULO
Published: April 2021

Analysis of gene expression in a mouse model of unilateral ureteral obstruction (UUO) revealed significant induction of CD14 mRNA in kidneys with obstructed ureters. Immunocytochemical analysis indicated that CD14 was upregulated in tubular epithelial cells and this upregulation was not attributable to infiltration of the kidneys by mononuclear cells. This induction of CD14 mRNA was found to occur in BALB/C, C57BL/6, C3H/HeN, and C3H/HeJ mice during UUO. Ischemia/reperfusion of kidneys also induced CD14 mRNA. Mice lacking either of the tumor necrosis factor-α receptor (TNFR) genes were also studied; the induction of CD14 was blunted in TNFR 1-knockout mice but not in TNFR2-knockout mice. Apoptosis of tubular cells in lipopolysaccharide-resistant CH3/HeJ mice was significantly (P < 0.05) less than that in lipopolysaccharide-responsive CH3/HeN mice during UUO. These results suggest that CD14 is acutely induced in tubular epithelial cells in two mouse models of renal injury. This induction is regulated by tumor necrosis factor-α, through TNFR1. CD14 may participate in the apoptosis of tubular epithelial cells on a more chronic basis by activating a pathway that is absent or deficient in C3H/HeJ mice.
Mechanism of the Inhibitory Effect of OPB-9195 [(±)-2-Isopropylidenehydrazono-4-oxo-thiazolidin-5-ylacetanilide] on Advanced Glycation End Product and Advanced Lipoxidation End Product Formation
Authors: MIYATA TOSHIO , UEDA YASUHIKO , ASAHI KOICHI , IZUHARA YUKO , INAGI REIKO , SAITO AKIRA , VAN YPERSELE DE STRIHOU CHARLES , KUROKAWA KIYOSHI
Published: April 2021

The accumulation in uremic plasma of reactive carbonyl compounds (RCO) derived from both carbohydrates and lipids (“carbonyl stress”) contributes to uremic toxicity by accelerating the advanced glycation and lipoxidation of proteins. It was previously demonstrated that OPB-9195 [(±)-2-isopropylidenehydrazono-4-oxo-thiazolidin-5-ylacetanilide] inhibited the in vitro formation of advanced glycation end products (AGE) in uremic plasma. This study was designed to elucidate the mechanism of action of OPB-9195 by further delineating the AGE and advanced lipoxidation end product (ALE) precursors targeted by this drug. The inhibitory effects of OPB-9195 on the formation of two AGE (Nε-carboxymethyllysine and pentosidine) on bovine serum albumin incubated with various AGE precursors were examined. Inhibition of Nε-carboxymethyllysine and pentosidine formation with OPB-9195 was more efficient than with aminoguanidine. OPB-9195 also proved effective in blocking the carbonyl amine chemical processes involved in the formation of two ALE (malondial-dehyde-lysine and 4-hydroxynonenal-protein adduct). The efficiency of OPB-9195 was similar to that of aminoguanidine. When glucose-based peritoneal dialysis fluid was incubated in the presence of OPB-9195, a similar inhibition of AGE formation was observed. The direct effect of OPB-9195 on major glucose-derived RCO in peritoneal dialysis fluids was then evaluated. The effects of OPB-9195 could be accounted for by its ability to trap RCO. The concentrations of three major glucose-derived RCO (glyoxal, methylglyoxal, and 3-deoxy-glucosone) were significantly lower in the presence of OPB-9195 than in its absence. Aminoguanidine had a similar effect. In conclusion, OPB-9195 inhibits both AGE and ALE formation, probably through its ability to trap RCO. OPB-9195 might prove to be a useful tool to inhibit some of the effects of RCO-related uremic toxicity.
Increased Albumin and Fibrinogen Synthesis in Hemodialysis Patients with Normal Nutritional Status
Authors: GIORDANO MAURO , FEO PIERPAOLO DE , LUCIDI PAOLA , dePASCALE EMANUELA , GIORDANO GELSOMINA , INFANTONE LORENA , ZOCCOLO ANNA MARIA , CASTELLINO PIETRO
Published: April 2021

This study compared the rates of whole-body proteolysis and of albumin and fibrinogen synthesis of seven hemodialysis patients (HD) with those of seven normal matched control subjects (C). HD patients had a normal nutritional and inflammatory status and serum albumin levels >3.5 g/dl. Endogenous leucine flux, albumin and fibrinogen fractional synthesis rate (FSR), and absolute intravascular synthesis rate (ASR) of albumin and fibrinogen all were evaluated by a primed/continuous infusion of 5,5,5-D3-L-leucine. Plasma volume was determined by the Evans blue dye dilution method. Endogenous leucine flux was significantly increased in HD (2.64 ± 0.08 μmol/kg per min) compared with C (2.17 ± 0.07 μmol/kg per min, P < 0.05). Serum albumin concentrations were similar in HD and C. Plasma fibrinogen levels were significantly increased in HD compared with C (P < 0.05). Plasma volume was greater in HD than in C (P < 0.05). As a result, total intravascular pool of both albumin (141 ± 7 versus 114 ± 3 g/1.73 m2, P < 0.05) and fibrinogen (11.7 ± 1 versus 6.7 ± 0.5 g/1.73 m2, P < 0.05) were greater in HD than in C. Albumin FSR was not statistically different in HD and C. However, albumin ASR was significantly increased in HD than in C (13.7 ± 2 versus 10.3 ± 1 g/1.73 m2 per d, P < 0.05). Similarly, FSR of fibrinogen did not differ in HD and C groups, whereas ASR of fibrinogen was significantly higher in HD than in C (3.31 ± 0.6 versus 1.94 ± 0.3 g/1.73 m2 per d, P < 0.05). In summary, normoalbuminemic HD patients have an increased intravascular pool with a greater absolute synthesis rate of both albumin and fibrinogen and an increased rate of whole-body leucine flux.
Limitations in Anthropometric Calculations of Total Body Water in Patients on Peritoneal Dialysis
Authors: JOHANSSON ANN CATHRINE , SAMUELSSON OLA , ATTMAN PER OLA , BOSAEUS INGVAR , HARALDSSON BO[Combining Diaeresis]RJE
Published: April 2021

Having an accurate estimation of total body water (TBW) is essential for the evaluation of dialysis efficacy in peritoneal dialysis (PD) patients. In this study, TBW volumes were measured by tritium dilution (TBWTHO) in 165 PD patients and compared with TBW calculations according to the Watson formulas. An alternative anthropometric formula based on the present PD population was also developed and validated in an independent sample of 29 PD patients. Furthermore, the relation between TBWTHO and body surface area (BSA) according to the formula of Gehan was analyzed. Body composition was assessed by a four-compartment model, based on measurements of TBWTHO and total body potassium. Mean values of TBW by the Watson formulas were almost identical to TBWTHO, and the correlation coefficient for the relationship of calculated to measured volumes was 0.89 (P = 0.001). However, both anthropometric formulas—the Watson formulas as well as the new, alternative one—overestimated TBW in obese patients and vice versa in lean patients. Similarly, TBW was underestimated in patients who were overhydrated. The correlation coefficient between TBWTHO and BSA was 0.708 for males and 0.797 for females (P = 0.0001 for both). In obese patients, the relationship was even closer (r = 0.924 and 0.911, respectively). In conclusion, anthropometric formulas to calculate TBW showed a considerable intraindividual variability compared with measured values. This was related to body composition features, such as degree of obesity and hydration. In contrast, BSA correlated closely to TBW in obese individuals. TBW as estimated by anthropometric formulas must be analyzed with caution, especially in the very obese or very lean patient.
Lowering of Microalbuminuria in Diabetic Patients by a Sympathicoplegic Agent
Authors: STROJEK KRZYSZTOF , GRZESZCZAK WLADYSLAW , GÓRSKA JUTA , LESCHINGER MONIKA I. , RITZ EBERHARD
Published: April 2021

There is convincing evidence for a specific BP-independent effect of angiotensin-converting enzyme inhibitors and angiotensin II receptor blockers on albuminuria in glomerular disease. Because progression of glomerular disease is not consistently halted by these agents, there is a need to explore potential renoprotective effects of other drugs. Recent animal work documented that nonhypotensive doses of moxonidine, a sympathicoplegic agent, reduce albuminuria and development of glomerulosclerosis in a BP-independent manner. A randomized, crossover design was used to assess the human relevance of the experimental data in 15 normotensive, nonsmoking type 1 diabetic mellitus patients with good glycemic control (age, 37.3 ± 6.6 yr; 9 men/6 women; duration of diabetes, 23.6 ± 5.1 yr) with baseline urinary albumin excretion rates (AER) >20 μg/min in the run-in phase. AER was assessed in overnight timed urine collections. The patients were assigned to a 3-wk placebo and a 3-wk moxonidine (0.2 mg twice a day) period, respectively, in random order. This dose causes modest BP lowering in hypertensive individuals but does not affect BP in normotensive individuals. There was no significant effect on ambulatory BP (mean arterial pressure, 91.8 ± 7.1 mmHg in the third week of placebo and 91.1 ± 8.7 mmHg on moxonidine). There was a significant (P < 0.006) difference of the treatment effects between placebo and moxonidine, respectively, on AER; median AER at the end of the placebo period was 39.8 μg/min (range, 15.9 to 117 μg/min) versus 29.0 (range, 9.03 to 85.8 μg/min) at the end of the moxonidine period. The data document an antialbuminuric effect of nonhypotensive doses of moxonidine. Diminished sympathetic traffic to the kidney is the most plausible explanation for the finding.
Induction of Cyclooxygenase-2 in Thick Ascending Limb Cells by Adrenalectomy
Authors: VIO CARLOS P. , AN SHAO-JIAN , CÉSPEDES CARLOS , MCGIFF JOHN C. , FERRERI NICHOLAS R.
Published: April 2021

Adrenalectomized (ADX) and sham-operated rats received either dexamethasone (DEX) or vehicle. Renal tissue was used for morphologic analysis, assessment of cyclooxygenase-2 (COX-2) protein expression and mRNA accumulation, and quantitation of COX-2 activity. In untreated or shamoperated rats, COX-2 protein was observed in a subset of tubular epithelial cells (<2%), which were located mainly in the cortex. All COX-2-positive cells also expressed Tamm-Horsfall glycoprotein, a highly selective marker for thick ascending limb (TAL) cells. After ADX, >30% of TAL cells expressed COX-2 in a manner consistent with recruitment of COX-2-positive TAL cells toward the medulla. Treatment of ADX rats with DEX reduced the number of COX-2-positive cells to that observed in sham-operated or intact rats. COX-2 mRNA accumulation was increased by ADX and partially attenuated by treatment with DEX. Western blot analysis of cortical microsomes revealed a substantial increase in COX-2 expression in ADX rats, compared with ADX/DEX-treated, sham-operated, or intact rats. The increase in COX-2 protein expression was associated with a twofold increase in prostaglandin E2 formation by cortical microsomes obtained from ADX rats, compared with sham-operated rats. It is concluded that ADX induces expression of enzymatically active COX-2, such that expression occurs in the cortical TAL and proceeds in a defined pattern toward the outer medullary TAL. It is suggested that ADX induces expression of TAL cells that, in the basal state, do not express COX-2 protein.
The Membrane-Associated Guanylate Kinase Protein MAGI-1 Binds Megalin and Is Present in Glomerular Podocytes
Authors: PATRIE KEVIN M. , DRESCHER ANDREW J. , GOYAL MEERA , WIGGINS ROGER C. , MARGOLIS BEN
Published: April 2021

The transmembrane endocytic receptor glycoprotein 330/megalin (hereafter referred to as megalin) is localized to the apical membrane domain of epithelial cells, where it is involved in the uptake of proteins from extracellular sources. The cytoplasmic domain of megalin contains amino acid motifs that have the potential to bind to other proteins, which may influence its localization or function. The yeast two-hybrid system was used to search for proteins that bind to the cytoplasmic tail of megalin, and a protein fragment from a mouse embryonic cDNA library that contained a single PDZ domain was identified. This protein, which was named glycoprotein 330-associated protein (GASP), appears to be a truncated mouse counterpart of the human and rat proteins atrophin-1-interacting protein-1 and synaptic scaffolding molecule, respectively. The interaction of GASP with megalin is mediated by the PDZ domain of GASP binding to the DSDV motif found at the carboxyl-terminus of megalin. A mutant version of megalin that lacks the terminal valine is unable to bind to GASP, illustrating the PDZ domain-dependent interaction between these two proteins. A close homolog of GASP, i.e., membrane-associated guanylate kinase with inverted orientation-1 (MAGI-1), is more ubiquitous in its tissue distribution (including kidney) and is also able to specifically bind to megalin via its fifth PDZ domain. Immunofluorescence studies of adult kidney revealed that MAGI-1 is expressed in the glomerulus of the kidney, in a manner that parallels the expression of the podocyte-specific protein glomerular epithelial protein 1. Western analysis of endogenous MAGI-1 from glomerular preparations suggests that it is associated with the cytoskeleton and seems to be expressed in a different form, compared with cell line-derived endogenous MAGI-1. The association of megalin with MAGI-1 may allow the assembly of a multiprotein complex, in which megalin may serve a nonendocytic function in glomerular podocytes.
Acetylcholine Increases the Free Intracellular Calcium Concentration in Podocytes in Intact Rat Glomeruli via Muscarinic M5 Receptors
Authors: NITSCHKE ROLAND , HENGER ANNA , RICKEN SIGRID , MU[Combining Diaeresis]LLER VICTORIA , KO[Combining Diaeresis]TTGEN MICHAEL , BEK MARTIN , PAVENSTA[Combining Diaeresis]DT HERMANN
Published: April 2021

The effects of acetylcholine (ACh) on the free intracellular calcium concentration ([Ca2+]i) of microdissected glomeruli were investigated using fura-2 fluorescence digital imaging and two-photon confocal microscopy. ACh caused a concentration-dependent [Ca2+]i increases with an initial peak followed by a sustained plateau, which was suppressed by reduced extracellular Ca2+ concentrations. The [Ca2+]i plateau was not affected by the L-type Ca2+ channel blocker nicardipine, whereas gadolinium and lanthanum (both at 1 μM) blocked the plateau. Diphenylacetoxy-N-methylpiperidine methiodide (100 nM), an M3/M5 receptor antagonist, and pirenzepine (1 μM), an M1 receptor antagonist, completely inhibited the effect of ACh. [Ca2+]i measurements using two-photon excitation of fluo-3 and staining of the cells with calcein/acetoxymethyl ester, for observation of the capillary network together with the glomerular cells, showed that [Ca2+]i was increased in single podocytes. Immunohistochemical studies did not demonstrate M3 receptor expression in glomerular cells. M1 receptors could be detected only in the parietal sheet of Bowman's capsule, whereas M5 receptors were found only in podocytes. The data show that ACh increases [Ca2+]i in podocytes of intact glomeruli, most likely via muscarinic M5 receptors.
Novel Nonsense Mutation in the Na+/HCO3 - Cotransporter Gene (SLC4A4) in a Patient with Permanent Isolated Proximal Renal Tubular Acidosis and Bilateral Glaucoma
Authors: IGARASHI TAKASHI , INATOMI JUN , SEKINE TAKASHI , SEKI GEORGE , SHIMADZU MITSUNOBU , TOZAWA FUMIKO , TAKESHIMA YASUHIRO , TAKUMI TORU , TAKAHASHI TOSHIKAZU , YOSHIKAWA NORISHIGE , NAKAMURA HAJIME , ENDOU HITOSHI
Published: April 2021

Permanent isolated proximal renal tubular acidosis (pRTA) with ocular abnormalities is a systemic disease involving short stature, isolated pRTA, mental retardation, and ocular abnormalities. Kidney Na+/HCO3
- cotransporter (kNBC1) cDNA from peripheral lymphocytes from a patient with permanent isolated pRTA and bilateral glaucoma was screened, and a novel homozygous mutation, namely a cytosine-to-thymine transition at nucleotide 234, which resulted in the formation of a stop codon at codon 29, was identified. This homozygous mutation, Q29X, was identified in the unique 5′-end of the kNBC1 gene (SLC4A4) of the patient. Cosegregation of this Q29X mutation with the disease and heterozygosity in the parents of the affected patient were observed. The absence of this mutation in 156 alleles from 78 Japanese individuals indicates that this mutation is directly related to the disease and is not a common DNA sequence polymorphism. This nonsense mutation predicts a truncated kNBC1 protein that lacks the 1007 amino acids of the carboxyl-terminus, and the effect on kNBC1 cotransport activity is likely to be a loss of function. In contrast, the pancreatic Na+/HCO3
- cotransporter of the patient is not likely to be affected by this nonsense mutation. These results have implications for understanding the role of kNBC1 in the pathophysiologic processes of pRTA associated with ocular abnormalities and mental retardation.
Mutation Analysis of the Entire Replicated Portion of PKD1 Using Genomic DNA Samples
Authors: PHAKDEEKITCHAROEN BUNYONG , WATNICK TERRY J. , GERMINO GREGORY G.
Published: April 2021

The replicated portion of PKD1, which comprises nearly 70% of the length of the gene, is predicted to harbor at least 85% of the mutations present in affected autosomal dominant polycystic kidney disease type 1 pedigrees. The relative paucity of reported mutations involving this segment is attributable to the significant technical challenges posed by the genomic structure of the gene. Previous genomic DNA-based strategies were unable to evaluate exons 1 and 22 and relied on the use of 10- to 13-kb PCR products. In this report, a set of six novel primer pair combinations, which can be used with previously reported reagents to analyze all of the exons in the replicated region (exons 1 to 34), are described. No product is greater than 5.8 kb in length, and various primer combinations can be used to reduce this length in half. Using this approach, two new pathogenic mutations, four novel disease-associated missense substitutions, and six new normal variants were identified. These new reagents should prove useful to investigators interested in performing DNA testing for this disorder.
Role of p53-Dependent Activation of Caspases in Chronic Obstructive Uropathy: Evidence from p53 Null Mutant Mice
Authors: CHOI YEONG-JIN , MENDOZA LEONARDO , RHA SUK-JOO , SHEIKH-HAMAD DAVID , BARANOWSKA-DACA ELZBIETA , NGUYEN VINH , SMITH WAYNE C. , NASSAR GEORGE , SUKI WADI N. , TRUONG LUAN D.
Published: April 2021

Chronic obstructive uropathy (COU) created by unilateral ureteric ligation is associated with increased renal cell apoptosis and p53 expression. Genetically engineered mice were used to examine the role of p53 in renal cell apoptosis in COU and the involved molecular pathways. Obstructed kidneys in p53+/+, p53+/-, and p53-/- mice were examined at days 4, 7, 15, 20, and 30 for apoptosis, and mRNA were examined for p53, members of the bcl-2 family, the death receptor family, and the common effectors of apoptosis. Obstructed kidneys in p53+/- and p53-/- mice exhibited equal attenuation of tubular and interstitial cell apoptosis (70 and 50%, respectively), compared with p53+/+ mice. However, p53 gene deficiency did not confer complete protection from apoptosis. Obstructed kidneys from p53-/- mice did not express p53 mRNA, whereas those from p53+/- and p53+/+ mice displayed mild and marked increase in their expression, respectively. Obstructed kidneys in p53+/+, p53+/-, and p53-/- mice displayed upregulation of mRNA for members of the bcl-2 family and most of the death receptor family, except for a lower level of tumor necrosis factor receptor-1, TRAIL, and FAP in p53+/+ mice. Obstructed kidneys in p53-/- and p53+/- mice showed virtual absence of caspase 11 and marked attenuation of caspases 1 and 12, contrasted with their strong expression in p53+/+ kidneys. These data suggest that apoptosis in obstructed kidneys involves p53-dependent as well as p53-independent pathways. The p53-dependent pathway may involve activation of caspases 1, 11, and 12, whereas the p53-independent pathway may involve activation of members of the bcl-2 and death receptor families.
Glomerular Injury and Tubular Loss in Adriamycin Nephrosis
Authors: JAVAID BASIT , OLSON JEAN L. , MEYER TIMOTHY W.
Published: April 2021

Glomerular injury manifested by sustained proteinuria usually leads to tubule injury and reduction of the GFR. The current study explored the link between these processes in rats with adriamycin nephrosis. One group of nephrotic rats received a vasopressin V2 receptor blocker (V2X) from 4 to 16 wk after injection of adriamycin, whereas a second group received no treatment (NoRx). V2 receptor blockade increased urine volume without affecting protein excretion. At 16 wk, both groups of nephrotic rats exhibited a marked reduction in GFR in comparison with normal controls (V2X, 0.22 ± 0.19 ml/min; NoRx, 0.20 ± 0.11 ml/min; control, 1.23 ± 0.11 ml/min). Morphologic studies revealed that the majority of glomeruli in nephrotic rats were no longer connected to normal tubule segments (V2X, 81 ± 21%; NoRx, 85 ± 18%; control, 1 ± 2%). Glomeruli without tubules were not, however, globally sclerosed. Disruption of the glomerular tubular junction was associated with the presence of amorphous material separating damaged tubule cells from the basement membrane. Serial sections revealed that this material spread from extensive areas of adhesion between the glomerular tuft and capsule to invest the tubular neck. Reduction of the GFR was strongly correlated with the fraction of glomeruli not connected to normal tubules (r
2 = 0.82; P < 0.0001). V2 receptor blockade did not preserve renal function or structure. These findings suggest that local extension of glomerular injury to destroy the tubule neck is an important cause of loss of renal function in adriamycin nephrosis.
The Potential of Bone Marrow-Derived Cells to Differentiate to Glomerular Mesangial Cells
Authors: IMASAWA TOSHIYUKI , UTSUNOMIYA YASUNORI , KAWAMURA TETSUYA , ZHONG YU , NAGASAWA RYUJI , OKABE MASARU , MARUYAMA NAOKI , HOSOYA TATSUO , OHNO TSUNEYA
Published: April 2021

Bone marrow stem cells (BMC) develop into hematopoietic and mesenchymal lineages but have not been known to differentiate into glomerular cells. To investigate whether such differentiation is possible, a search was made for donor glomerular cells in lethally irradiated C57BL/6j (B6) mice given transplants of BMC from syngeneic mice transgenic for green fluorescence protein (GFP) ([GFP→B6] mice). After the recipients of donor BMC manifested GFP-positive cells in their glomeruli, the numbers of such cells increased markedly, in a time-dependent manner, from 2 wk to 24 wk after bone marrow transplantation. Immunohistochemical analyses revealed that most GFP-positive cells in the glomeruli were neither macrophages nor T cells. With the use of a laser-scanning confocal microscope, GFP-positive cells were observed within the mesangium of [GFP→B6] mice. Furthermore, indirect immunofluorescence assays demonstrated that desmin-positive cells in the glomeruli of these chimeric mice were also positive for GFP. Among glomerular cells isolated from [GFP→B6] mice 24 wk after bone marrow transplantation and then cultured, the majority of cells (approximately 84%) stained for desmin and approximately 60% of the desmin-positive cells expressed GFP. In addition, these GFP-positive cells in the cultures contracted in response to angiotensin II stimulation. These results suggest that bone marrow-derived cells may have the potential to differentiate into glomerular mesangial cells.
Impaired Angiogenesis in the Remnant Kidney Model
Authors: KANG DUK-HEE , HUGHES JEREMY , MAZZALI MARILDA , SCHREINER GEORGE F. , JOHNSON RICHARD J.
Published: April 2021

Impaired angiogenesis and decreased vascular endothelial growth factor (VEGF) expression were recently documented in the remnant kidney (RK) model of progressive renal failure. VEGF (50 μg/kg, twice daily) was administered to RK rats between weeks 4 and 8 after surgery, and rats were euthanized at week 8 for histologic study. During the administration of VEGF (n = 7) or vehicle (n = 6), systemic BP was comparable in the two groups. VEGF treatment resulted in improved renal function and lower mortality rates, compared with the vehicle-treated group. Renal histologic analyses confirmed a 3.5-fold increase in glomerular endothelial cell proliferation (0.14 ± 0.03 versus 0.04 ± 0.02 proliferating endothelial cells/glomerulus, VEGF versus vehicle, P < 0.05), a twofold increase in peritubular capillary endothelial cell proliferation (1.60 ± 0.30 versus 0.78 ± 0.17 cells/mm2, VEGF versus vehicle, P < 0.01), a threefold decrease in peritubular capillary rarefaction (P < 0.01), and a twofold increase in endothelial nitrix oxide synthase expression (P < 0.05) in the VEGF-treated group; an eightfold increase in urinary nitrate/nitrite levels (P < 0.05) was also noted. Although the difference in glomerulosclerosis scores did not reach statistical significance (0.67 ± 0.42 versus 1.22 ± 0.63, VEGF versus vehicle; range, 0 to 4; P = NS), VEGF-treated rats exhibited less interstitial collagen type III deposition (9.32 ± 3.26 versus 17.45 ± 7.50%, VEGF versus vehicle, P < 0.01) and reduced tubular epithelial cell injury, as manifested by osteopontin expression (5.57 ± 1.60 versus 9.58 ± 3.45%, VEGF versus vehicle, P < 0.01). In conclusion, VEGF treatment reduces fibrosis and stabilizes renal function in the RK model. The use of angiogenic factors may represent a new approach to the treatment of kidney disease.
Response to Single and Divided Doses of Shiga Toxin-1 in a Primate Model of Hemolytic Uremic Syndrome
Authors: SIEGLER RICHARD L. , PYSHER THEODORE J. , TESH VERNON L. , TAYLOR FLETCHER B.
Published: April 2021

Postdiarrheal hemolytic uremic syndrome is caused by Shiga toxin (Stx)-producing Escherichia coli. It was shown previously that the baboon, like the human, has glycolipid receptors for Stx in the gut and the kidney and that a single 50- to 200-ng/kg intravenous dose of purified Stx-1 results in thrombocytopenia, hemolytic anemia, and renal thrombotic microangiopathy. For further characterization of factors that modulate disease expression, the baboon's response to the intravenous administration of 100 ng/kg Stx-1 given either rapidly as a single bolus or slowly as four 25-ng/kg doses at 12-h intervals was compared. Animals that received the Stx-1 as a single dose developed thrombocytopenia, schistocytosis, and acute renal failure. Urinary but not plasma tumor necrosis factor-α concentrations rose significantly by 6 h and then declined rapidly. Urinary and plasma interleukin-6 concentrations rose later. Glomeruli showed reduced patency of capillary loops, fragmented red blood cells, fibrin and platelet microthrombi, necrosis and detachment of endothelial cells, and accumulation of flocculent material in subendothelial spaces. Damage to tubular epithelium and peritubular capillary endothelium also was seen. Animals that received four divided doses of Stx-1 developed no clinical or histologic features of hemolytic uremic syndrome. It is concluded that in the primate model, disease expression is modulated by the rate of Stx administration, and it is speculated that in the human, the rate of Stx absorption from the gut is one determinant of disease severity.
Effect of Ibandronate on Bone Loss and Renal Function after Kidney Transplantation
Authors: GROTZ WOLFGANG , NAGEL CHRISTIAN , POESCHEL DARIA , CYBULLA MARKUS , PETERSEN KARL-GEORG , UHL MARKUS , STREY CHRISTOPH , KIRSTE GU[Combining Diaeresis]NTER , OLSCHEWSKI MANFRED , REICHELT ACHIM , RUMP LARS CHRISTIAN
Published: April 2021

Severe osteoporosis frequently is observed after organ transplantation. In kidney transplantation, it adds to pre-existing renal bone disease and strategies to prevent osteoporosis are not established. Eighty kidney recipients were included in a randomized controlled prospective intervention trial. Treated patients (n = 40) received an injection of ibandronate, a bisphosphonate, immediately before and at 3, 6, and 9 mo after transplantation. The primary outcome measured was the change in bone mineral density. Secondary measures included graft outcome, spinal deformities, fracture rate, body height, and hormonal and metabolic data. Loss of spongy and cortical bone after transplantation was prevented by ibandronate. Changes of bone mineral density (ibandronate versus controls) were as follows: lumbar spine, -0.9 ± 6.1% versus -6.5 ± 5.4% (P < 0.0001); femoral neck, +0.5 ± 5.2% versus -7.7 ± 6.5% (P < 0.0001); and midfemoral shaft, +2.7 ± 12.2% versus -4.0 ± 10.9% (P = 0.024). Fewer spinal deformities developed with ibandronate (7 patients with 7 deformities versus 12 patients with 23 deformities; P = 0.047). Loss of body height was 0.5 ± 1.0 cm versus 1.1 ± 1.0 cm in control subjects (P = 0.040). Two bone fractures occurred in each group. There were fewer acute rejection episodes with ibandronate (11 versus 22; P = 0.009). Graft function after 1 yr was comparable. Bone loss, spinal deformation, and loss of body height during the first year after kidney transplantation are prevented by injection of ibandronate at intervals of 3 mo. The smaller number of rejection episodes of the ibandronate-treated group should be confirmed and its mechanism should be explored in additional studies.