Cytotoxicity evaluation, antibacterial effect, and degree of conversion of QAM-containing adhesives

Abstract The aim of this study was to evaluate the influence of adding quaternary ammonium methacrylates (QAMs) to experimental adhesives by assessing the degree of conversion (DC), cytotoxicity against keratinocytes and fibroblasts, and antibacterial activity against biofilm formation. Two QAMs were added to an experimental adhesive: dimethylaminododecyl methacrylate bromododecane (DMADDM) or dimethylaminododecyl methacrylate bromohexadecane (DMAHDM) at three concentrations each: 1, 2.5, and 5 wt.%. Experimental adhesive without QAMs (control group) and commercially available Transbond XT Primer (3M Unitek, Monrovia, California, USA) were used for comparisons. The adhesives were tested for DC, cytotoxicity against keratinocytes and fibroblasts, and antibacterial activity against biofilm formation. DC, cytotoxicity against fibroblasts, and antibacterial activity were analyzed using one-way ANOVA and Tukey's multiple comparisons. Cytotoxicity against keratinocytes was evaluated using the Kruskal Wallis and Dunn's post-hoc (α = 5%) tests. Transbond showed lower DC as compared to 5% DMAHDM, 1% DMADDM, and 5% DMADDM (p < 0.05). However, all groups presented proper DC when compared to commercial adhesives in the literature. In the evaluation of cytotoxicity against keratinocytes, Transbond induced higher viability than 2.5 wt.% groups (p < 0.05). Against fibroblasts, Transbond induced higher viability as compared to 5 wt.% groups (p < 0.05). DMAHDM at 5 wt.% reduced biofilm formation when compared to all the other groups (p < 0.05). Despite their cytotoxic effect against keratinocytes, gingival fibroblasts showed higher viability. DMAHDM at 5 wt.% decreased Streptococcus mutans viability. The incorporation of DMAHDM at 5 wt.% may be a strategy for reducing the development of white spot lesions.

A single-center clinical study of 61 children with ammonium urate stones / 现代泌尿外科杂志

【Objective】 To analyze the clinical characteristics of children with ammonium urate stones in Xinjiang, so as to provide reference for the prevention and treatment of this disease. 【Methods】 The clinical data of all children with ammonium urate stones admitted to the People’s Hospital of Xinjiang Uygur Autonomous Region from 2016 to 2021 were retrospectively analyzed, including age, sex, body mass index, stone site, stone size, stone component, urine pH, urine culture and biochemical examination results. The serum total protein, albumin, sodium, potassium, calcium, magnesium, uric acid and urine pH were compared between the pure and mixed groups. 【Results】 A total of 61 children (31.6%) had ammonium urate stones, their average age was (4.05±3.37) years, and the male to female ratio was 2.21∶1. Among them, there were 37 cases (60.7%) of renal calculi and 50 cases (82.0%) of upper urinary calculi. The most common component of mixed ammonium urate stones was calcium oxalate, including calcium oxalate monohydrate, calcium oxalate monohydrate and calcium oxalate dihydrate. Compared with mixed type, children with pure stone type had a younger age (P=0.001) and a smaller stone size (P=0.003). Positive urine culture was detected in 14 cases (23.0%), 7 of which (50% were infected with Escherichia coli, and 11 (78.6%) with non-urease bacteria. 【Conclusion】 Non-urease bacteria are the main pathogens of urinary tract infection in children with ammonium urate stones. The incidence is higher in boys, and the most common stone location is upper urinary tract. Calcium oxalate is the most common mixed component. Pure type is more common in young children and the stones are relatively small.

Research progress of bacterial biofilm eradication agents based on modifications of natural products / 药学学报

Bacterial biofilm widely exists in all kinds of bacteria, and is related to about 80 percent of bacterial infections. It is one of the main reasons for bacterial tolerance and resistance to existing antibiotics. Therefore, there is unmet clinical need for new anti-biofilm drugs. At present, there are three kinds of anti-biofilm agents under research, including biofilm inhibitors, biofilm dispersal agents and biofilm eradication agents. Among them, the biofilm eradication agent is unique, which can not only kill bacteria in biofilm but also eliminate biofilm as a monotherapy. Based on modifications of natural products with antibacterial activity, a variety of compounds with biofilm eradicating activities have been obtained, such as, acyldepsipeptides, pyrrolomycins, halogenated phenazines and halogenated 8-hydroxyquinolines. In this review, we summarize several major biofilm eradication agents above according to their structures and mechanisms.

Research progress on root canal irrigation disinfection drugs / 口腔疾病防治

@#Endodontic infection control is crucial to successful root canal treatment. Irrigation is the key step in endodontic procedures, and the application of root canal irrigation and disinfection medications play an important role. How to enhance antibacterial effects and functions in removing tissues while maintaining biocompatibility is a hot topic in endodontics. Currently, insights to address this issue can be split into two categories: one, the modification or combination of conventional endodontic irrigation solutions, and two, the development of novel endodontic irrigation solutions with new technologies and materials, for instance, nanomaterials and natural exacts. However, conventional endodontic irrigation solutions, such as sodium hypochlorite and chlorhexidine, are still the first choice in clinical practice. Most novel endodontic irrigation solutions remain at the pre-clinical laboratory stage. Clinical research and relevant data are required to determine whether various methods can improve endodontic irrigation. From basic research to clinical application is the direction for advancing to the next stage. The present article focuses on research progress on endodontic irrigation, especially concerning its antibacterial mechanism, characteristics and efficacy, to provide a reference for future clinical translation.

Establecimiento de biofloc a tres relaciones carbono/nitrógeno, tendiente a la producción de zooplancton / Establishment of biofloc at three carbon/nitrogen ratios, tending to the production of zooplankton

RESUMEN El objetivo de esta investigación fue establecer el biofloc a tres relaciones carbono/ nitrógeno (C/N): 10/1, 15/1 y 20/1, determinando la secuencia de remoción de N, el perfil de sólidos y la caracterización del zooplancton, para tal fin se dispuso de tres tanques con volumen de 7000 L, incorporando oxígeno al agua a través de un aireador tipo soplador. Se utilizó como fuente de N balanceado, fuente de carbono melaza y bicarbonato de sodio como fuente alcalinizante. Al inicio se incrementó el nitrógeno amoniacal total NAT a 2 mg/L, la alcalinidad total (AT) a 120 mg/L y se adicionó como inóculo 10 litros/tanque de agua proveniente de un estanque de cultivo, al sexto y décimo días se adicionó balanceado incrementando teóricamente el NAT en 4 mg/L y a partir del día 12 en 1 mg/L. En las tres relaciones C/N se evidenciaron procesos de nitrificación durante la estabilización del biofloc, hasta llegar en el tiempo a concentraciones no letales de amonio y nitrito para peces, menores a 1 mg/L. En cuanto a los sólidos volátiles, se encontró una mayor concentración en la relación 20/1, lo cual puede atribuirse a la mayor adición de melaza, con la consecuente producción de SSV a partir de la dominancia de comunidades heterotróficas, en los tres macrocosmos se presentaron comunidades del zooplancton, no obstante, el T2 presentaron la mayor abundancia y riqueza de organismos. Las tres relaciones C/N en biofloc establecieron condiciones de calidad de agua y alimento vivo.

ABSTRACT The objective of this research was to establish the biofloc at three carbon/nitrogen (C/N) relationships: 10/1, 15/1 and 20/1, determining the N removal sequence, the solids profile and the characterization of the zooplankton, for this purpose three tanks with a volume of 7000 L were available, incorporating oxygen into the water through a blower-type aerator. It was used as a source of balanced N, a source of carbon molasses and sodium bicarbonate as an alkalizing source. At the beginning, the total ammoniacal nitrogen NAT was increased to 2 mg/L, the total alkalinity (AT) to 120 mg/L and 10 liters / tank of water from a culture pond was added as inoculum, on the sixth and tenth days it was he added balanced, theoretically increasing the NAT by 4 mg/L and from day 12 by 1 mg/L. In the three C / N relationships, nitrification processes were evidenced during the stabilization of the biofloc, until reaching non-lethal concentrations of ammonium and nitrite for fish, less than 1 mg/L in time. Regarding volatile solids, a higher concentration was found in the 20/1 ratio, which can be attributed to the greater addition of molasses, with the consequent production of SSV from the dominance of heterotrophic communities, in the three macrocosms there were Zooplankton communities, however, T2 presented the highest abundance and richness of organisms. The three C / N relationships in biofloc established conditions of water quality and live food.

Determination of arsenic in urine by hydride generation atomic fluorescence spectrometry with ammonium persulfate digestion / 中华地方病学杂志

Objective:To establish a hydride generation atomic fluorescence method using ammonium persulfate as the digestion reagent for determination of arsenic in urine (hereinafter referred to as this method).Methods:The collected urine samples with ammonium persulfate were heated and digested on the tubular electric heating automatic control constant temperature digester (60 holes), with 5% hydrochloric acid solution as reaction medium and current carrier and 1.5% potassium borohydride solution as reducing agent. Arsenic content was determined with a four-channel atomic fluorescence spectrometer. The arsenic standard solution of 0 - 10 μg/L was prepared to determine the standard curve of this method, and the method was evaluated from the detection limit, linear range, correlation coefficient, precision, standard addition recovery experiment, and urine arsenic quality control sample detection. The standard method "Determination of Arsenic in Urine by Hydride Generation Atomic Fluorescence Spectrometry" (WS/T 474-2015, referred to as the standard method) was used for comparison experiments.Results:When the sampling volume was 1 ml, the detection limit of this method (digest with 1 ml 1.5 mol/L ammonium persulfate) was 0.03 μg/L. In the range of arsenic content from 0 - 10 μg/L, the linear relationship between arsenic content and fluorescence intensity was good, and the correlation coefficients ( r) were all 0.999 9. The relative standard deviations( RSD) of the three replicates of urine samples with different concentrations were 1.00%, 0.89% and 0.49%, respectively. Urine arsenic quality control samples were tested, and the test results were all within the range of public values; the overall average recovery was 102.29%, and the recovery range was 92.10% - 108.15%. Compared with the standard method in the determination results of 20 urine samples, the difference was not statistically significant ( t = - 0.40, P > 0.05). Conclusions:The hydride generation atomic fluorescence spectrometry using ammonium persulfate as digestion reagent for the determination of arsenic in urine has the advantages of low detection limit, good precision, high accuracy, small amount of sampling and digestion reagent, simple operation, and less harmful gas generation in sample pretreatment. It is suitable for rapid determination of arsenic in urine in large quantities.

Evaluation of current disinfection practice of transvaginal ultrasound probes in a Philippine Tertiary Referral Hospital: A comparative study on the performance of manual reprocessing methods

Objectives@#There are no standard infection control regulations in transvaginal ultrasound probe disinfection followed in the most prominent local public tertiary referral hospital. Likewise, no studies have evaluated the efficacy of the current method that uses an inexpensive multipurpose antiseptic spray solution. This study aims to evaluate the efficacy of the current practice of manual disinfection of TVS probes and compare it with the performance of an acceptable manual reprocessing method. @*Methods@#A prospective, randomized, controlled study was carried out using a crossover, quasi-experimental design, collecting 119 total samples from the ultrasound transducers before (35 samples) and after disinfection with two manual reprocessing methods, either a locally manufactured multipurpose antiseptic spray (A-Septic® Multipurpose Antiseptic Spray) that is currently used for disinfection or Mikrozid Sensitive®, a ready to use impregnated wipes (42 samples each arm). Disinfection efficacy was evaluated based on microbial culture results. @*Results@#Before disinfection, bacterial growth was observed in 77.1% (27/35) of the probes. After disinfection, 80.95% (34/42) remained contaminated with the antiseptic spray and 21.43% (9/42) with the wipes. The cultures revealed many environmental and pathogenic bacterial isolates, including Burkholderia, Staphylococcus, Acinetobacter, Diphtheroids, and Pseudomonas. @*Conclusions@#The currently used method for disinfecting transvaginal transducers in the division is not adequate for decontamination and decreasing the risk of cross contamination among patients. The results call for aggressive disinfection measures and highlight the need to update local standards and formulate and institutionalize these recommendations.

Resistencia antimicrobiana y daño al medio ambiente, en el contexto COVID-19: Una revisión sistemática / Antimicrobial resistance and damage to the environment, in the context of COVID-19: A systematic review

Las pautas de abordaje de los pacientes con COVID-19, al inicio de la pandemia, se realizó por ensayo, ya que se desconocía la fisiopatología de esta nueva enfermedad, entre las acciones médicas se describió el uso de antiboticoterapias de manera indiscriminada, también se instó a realizar desinfección profundas, muchas veces, con sustancias químicas con impacto negativo en la salud ambiental, tanto a nivel de la microbiota encargada del equilibrio ecológico, como en la contaminación ambiental, principalmente del agua. Aunado a esto, debido a la emergencia sanitaria, se requirió usar en mayor cantidad equipos de protección personal de un solo uso y, como consecuencias el aumento de desechos sólidos peligrosos, cuya composición son de degradación tardía. Por tanto, la respuesta sanitaria ante la pandemia, probablemente fue escenario para acentuar la resistencia antimicrobiana y el riesgo de daño ambiental. En este, artículo se realizó una revisión sistemática de la literatura científicas en torno a estos tópicos, las evidencias demostraron el aumento de mecanismos de resistencias de bacterias, principalmente, patógenas del tracto respiratorio. De igual manera, el impacto negativo por uso irracional de desinfectantes químicos, traducido en bacterias resistentes especialmente a compuestos de amonio cuaternario. La bioacumulación y biomagnificación de estas sustancias ha provocado toxicidad, mutaciones, propagación de genes de resistencia. Por lo tanto, se sugiere que se priorice las estrategias que mitiguen el rastro que ha venido extendiéndose al paso de la pandemia por el SAR- CoV-2(AU)

The guidelines for the approach of patients with COVID-19, at the beginning of the pandemic, were carried out by trial, since the pathophysiology of this new disease was unknown, among the medical actions the use of antibiotic therapies indiscriminately was described, also the urged to carry out deep disinfection, many times, with chemical substances with a negative impact on environmental health, both at the level of the microbiota responsible for the ecological balance, and in environmental pollution, mainly of water. In addition to this, due to the health emergency, it was required to use more single-use personal protective equipment and, as a consequence, the increase in hazardous solid waste, whose composition is delayed degradation. Therefore, the health response to the pandemic was probably the setting to accentuate antimicrobial resistance and the risk of environmental damage. In this article, a systematic review of the scientific literature on these topics was carried out, the evidence demostrated the increase in resistance mechanisms of bacteria, mainly pathogens of the respiratory tract. Anyways, the negative impact due to the irrational use of chemical disinfectants, translated into resistant bacteria, especially to quaternary ammonium compounds. The bioaccumulation and biomagnification of these substances has caused toxicity, mutations, propagation of resistance genes. Therefore, it is suggested that strategies be prioritized that mitigate the trail that has been spreading as the pandemic passed by SAR-CoV-2(AU)

Use of ammonium chloride to prevent urolithiasis in sheep / [Utilização do cloreto de amônio na prevenção de urolitíase em ovinos]

Urolithiasis has a high incidence among confined sheep. It is multifactorial and may cause economic damage. Our aim was to determine the capacity of urinary acidification using ammonium chloride in sheep. Twenty-five 3-month-old male sheep were confined and randomly divided into three groups; the G200 and G500 groups received 200mg/kg/GW and 500mg/kg/GW of ammonium chloride daily for 56 consecutive days, respectively, whereas the CG group did not receive ammonium chloride. Sampling times and clinical evaluation were performed weekly, starting from the 14th day of confinement (M1 or immediately before administering ammonium chloride) until the 17th day (M9) of the feedlot. Hemogasometry, biochemical examination of serum urea and creatinine concentration and ultrasound evaluation of the urinary tract were performed. The urinalysis indicated a higher incidence of ammonium magnesium phosphate crystals at the beginning of the study, showing a migration to urate crystal formation, mainly in the G500 group because of urinary acidification. No hemogasometric, serum biochemistry, ruminal fluid, or ultrasonographic changes were observed. Urinary acidification was achieved and maintained after M7 during the administration of ammonium chloride in the G500 group, but not in the other study groups.(AU)

A urolitíase apresenta alta incidência em ovinos confinados, etiologia multifatorial, e pode causar prejuízo econômico. O objetivo do presente estudo foi determinar a capacidade da acidificação urinária mediante o uso de cloreto de amônio em ovinos. Foram utilizados 25 ovinos de três meses de idade, confinados e divididos aleatoriamente em três grupos: grupo CG (controle) não recebeu cloreto de amônio; grupo G200 (200mg/kg/PV) recebeu cloreto de amônio por 56 dias consecutivos; grupo G500 (500mg/kg/PV) recebeu cloreto de amônio por 56 dias consecutivos, administrados diariamente por via oral. Os momentos (M) de colheita de amostras e de avaliação clínica foram realizados com intervalo de sete dias, sendo M1 (imediatamente antes do cloreto de amônio), M2 (sete dias após) até M9, totalizando 70 dias de confinamento. Foram realizadas hemogasometria, concentração sérica de ureia e creatinina e avaliação ultrassonográfica do trato urinário. Na urinálise, houve uma maior incidência de cristais de fosfato amônio magnesiano no início do estudo, com migração para formação de cristais de urato, principalmente no G500, devido à acidificação urinária. Não houve alterações hemogasométricas, na bioquímica sérica, no líquido ruminal, ou alterações ultrassonográficas. A acidificação urinária foi obtida e mantida a partir do M7 durante a administração do cloreto de amônio no grupo G500, não ocorrendo nos outros grupos de estudo.(AU)

Polifluoruro de amonio como alternativa de acondicionamiento para laminados ultradelgados de disilicato de litio. Reporte de caso / Ammonium polyfluoride as an alternative for the surface conditioning of ultrathin lithium disilicate veneers. A case report

RESUMEN: Los laminados vitrocerámicos ultradelgados constituyen una alternativa conservadora para la resolución de alteraciones estéticas. Sin embargo, su acondicionamiento con ácido fluorhídrico suele no estar exento de complicaciones, por lo que el uso de un sistema autograbante en base a polifluoruro de amonio (Monobond Etch&Prime, MEP) permite disminuir el riesgo de sobregrabado, simplificando la técnica. El presente reporte presenta el seguimiento a dieciocho meses de un caso clínico resuelto mediante laminados vitrocerámicos acondicionados únicamente con MEP. Caso. Paciente joven con alteraciones estéticas en el sector anterosuperior. Se realizaron preparaciones conservadoras para la confección de laminados ultradelgados en disilicato de litio (e.maxPress). Las vitrocerámicas fueron acondicionadas sólo con MEP y se cementaron con Variolink Esthetic LC. Al año y medio las restauraciones se observaron indemnes, manteniendo un buen ajuste marginal y ausencia de tinciones. Conclusiones. MEP aparece como una alternativa prometedora para reemplazar al ácido fluorhídrico en la cementación de laminados vitrocerámicos.

ABSTRACT: Ultrathin glass-ceramic veneers are a conservative approach for the restorative treatment of aesthetic disorders in the anterior region. However, surface conditioning with hydrofluoric acid of the thin structures can be usually a challenging step. Therefore, the use of a self-etching ceramic primer containing ammonium polyfluoride (Monobond Etch&Prime, MEP) offers a simplified technique with a reduced risk of overetching. Here we present an 18-month follow-up of ultrathin glass-ceramic veneers conditioned with MEP only. Case. Young female patient with an aesthetic disorder in the front region. The four upper incisors were conservatively prepared for ultrathin lithium disilicate (e.maxPress) veneers. Pre-treatment of the glass-ceramics consisted only of MEP application, followed by adhesive cementation with Variolink Esthetic LC. At the 1.5-year recall, the restorations appeared undamaged, with no marginal gap or staining. Conclusions. MEP constitutes a promising alternative to hydrofluoric acid for the adhesive cementation of glass-ceramic veneers.

Physicochemical properties and biological effects of quaternary ammonium methacrylates in an experimental adhesive resin for bonding orthodontic brackets

Abstract Fixed orthodontic appliances may lead to biofilm accumulation around them that may increase caries risk. This study aimed to evaluate the influence of quaternary ammonium methacrylates (QAMs) on the physicochemical properties, cytotoxicity, and antibacterial activity of adhesive resins for orthodontic purposes. Methodology: A base resin was prepared with a comonomer blend and photoinitiator/co-initiator system. Two different QAMs were added to the base adhesive: dimethylaminododecyl methacrylate at 5 wt.% (DMADDM) or dimethylaminohexadecyl methacrylate (DMAHDM) at 10 wt.%. The base adhesive, without QAMs, (GC) and the commercial Transbond™ XT Primer 3M (GT) were used as control. The resins were tested immediately and after six months of aging in the water regarding the antibacterial activity and shear bond strength (SBS). The antibacterial activity was tested against Streptococcus mutans via metabolic activity assay (MTT test). The groups were also tested for the degree of conversion (DC) and cytotoxicity against keratinocytes. Results: The resins containing QAM showed antibacterial activity compared to the commercial material by immediately reducing the metabolic activity by about 60%. However, the antibacterial activity decreased after aging (p<0.05). None of the groups presented any differences for SBS (p>0.05) and DC (p>0.05). The incorporation of DMADDM and DMAHDM significantly reduced the keratinocyte viability compared to the GT and GC groups (p<0.05). Conclusion: Both adhesives with QAMs showed a significant reduction in bacterial metabolic activity, but this effect decreased after water aging. Lower cell viability was observed for the group with the longer alkyl chain-QAM, without significant differences for the bonding ability and degree of conversion. The addition of QAMs in adhesives may affect the keratinocytes viability, and the aging effects maybe decrease the bacterial activity of QAM-doped materials.

Improved synthetic process of apremilast / 中国药科大学学报

@#To optimize the process of hydrogenation reduction in the synthesis of apremilast (APST), 3-nitrophthalic anhydride (4) and (S)-1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethanamine-(S)-2-acetamido-4-methylpentanoate (7) were used as starting materials to synthesize (S)-2-(1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl)ethyl)-4-nitroisoindoline-1,3-dione (8) by amination.Then compound 8 was reduced to (S)-4-amino-2-(1-(3-ethoxy-4-methoxyphenyl)-2-(methylsulfonyl) ethyl) isoindoline-1,3-dione (9) with ammonium formate as hydrogen source and palladium hydroxide as catalyst.Finally, apremilast was obtained by the acetylation reaction with acetic anhydride.The structure of the products were verified by optical rotation, 1H NMR, 13C NMR, MS and elemental analysis.And the total yield of three steps was increased to 67.0%.The improved reduction process can avoid the special reaction of hydrogenation and pressurization, and reduce the safety risk and production costs with high commercial value.

Action of benzalkonium chloride in different pH

Ammonium quaternary compounds are widely used in poultry and swine production as disinfectants in the control of pathogens. They act on gram-positive bacteria, gram-negative bacteria, enveloped fungi and viruses. However, in some conditions of pH and presence of organic matter can be inactivated. This study evaluated the action of ammonium quaternary compounds at 1:1,000 and 1:2,000 dilutions against Salmonella enterica serovarTyphimurium and Salmonella enterica serovar Enteritidis in the presence of three different organic matter simulators, fetal bovine serum, skim milk and whole milk concentration of 1, 3, 5, and 7% and at pH 6 and 9, with 15 min of contact. It was possible to verify that the organic matter simulators adjusted in the same conditions of contact time and percentage, in the in vitro tests, presented different results and the fetal bovine serum did not inactivate the disinfectant. However, the best result against S. Typhimurium and S. Enteritidis was obtained at pH 6 at the dilution of 1:1,000 in all organic matter simulators.

Antagonistic microorganisms and nitrogen fertilization in control of tomato southern blight

The present study assessed the efficacy of formulated biocontrol agents and nitrogen fertilization on southern blight control. Antagonism test in vitro was performed to assess the inhibitory activity of Bacillus methylotrophicus and Trichoderma asperellum against the growth of Sclerotium rolfsii. Tomato seedlings were transplanted into the substrate added with ammonium nitrate doses and inoculated with the formulated biocontrol agents Ônix (B. methylotrophicus) or Quality (T. asperellum). Subsequently, seedlings were inoculated with S. rolfsii. Plant mortality, shoot and root weight were assessed 11 days after the last inoculation. Agents had effective inhibitory activity against S. rolfsii; thus, they could reduce southern blight severity when combined with ammonium nitrate. However, plant mortality was not reduced by them.(AU)

Comparison of methods for enriching urine proteins / 生物工程学报

The abundance of proteins in human urine is low and easily to be masked by high-abundance proteins during mass spectrometry analysis. Development of efficient and highly selective enrichment methods is therefore a prerequisite for achieving deep coverage of urine protein markers. Notably, different experimental methods would affect the urine protein enrichment efficacy and the coverage of urine proteome. In this study, ultrafiltration, nitrocellulose membrane enrichment and saturated ammonium sulfate precipitation were used to process 10 mL urine samples from five healthy volunteers and five bladder cancer patients. The urine proteins were enriched and separate by SDS-PAGE to compare the purification efficiency of different methods. Moreover, the peptide identification effects of different purification methods were analyzed by mass spectrometry to determine the best method for enriching urine protein histones. Saturated ammonium sulfate precipitation method outperformed the ultrafiltration and the nitrocellulose membrane enrichment methods in terms of the protein enrichment efficacy and quality. The interference of highly abundant albumin was reduced, whereas the amount of low-abundance protein was increased, and the sensitivity of mass spectrometry identification was increased. The saturated ammonium sulfate precipitation method may be applied for large-scale urine processing for screening clinical diagnostic markers through proteomics.

Extraction, Partial Purification and Characterization of Bromelain from Pineapple (Ananas Comosus) Crown, Core and Peel Waste

Abstract Ananas Comosus (also known as pineapple) is a part of Bromeliaceae family and it is consumed as food as well as folk medicine for the treatment of various diseases. It is reported that pineapple is a rich source of bromelain, a cysteine protease and it is considered as an important enzyme in different industries due to its significant therapeutic and industrial applications such as anticancer, anti-inflammatory and meat tenderizing. Bromelain is mostly present in fruit and stem of pineapple, but it is reported that crown, core, and peels, which constitute the waste of the pineapple plant, also contain bromelain but limited data is available. Therefore, the proposed study aimed at utilizing pineapple waste for the extraction and characterization of bromelain. Firstly, crude bromelain was extracted with phosphate buffer (pH 7), then it was subjected to partial purification using different fractions of ammonium sulphate (NH4)2SO4 such as 30, 40, 50 and 60% followed by desalting and concentration. Enzyme activity was calculated by using casein digesting unit (CDU) method. The results demonstrated that the crown bromelain showed highest purification of 4.34-fold at 30% (NH4)2SO4 saturation, whereas core and peel bromelain showed highest purification of 2.75 and 2.59-fold at 40% (NH4)2SO4 saturation. The molecular weight of crude and partially purified bromelain was determined by SDS-PAGE analysis and found to be 26 KDa. The pH and thermal stability of all the parts of pineapple showed maximum stability at pH 7 and at 35oC temperature.

Prepartum anionic diet induces hyperchloremic acidosis in high-producing dairy cows without preventing subclinical hypocalcemia / Dieta aniônica pré-parto provoca acidose hiperclorêmica em vacas leiteiras de alta produção mas não evita a hipocalcemia subclínica

In this study we evaluated the effects of the prepartum anionic diet on the electrolyte balance and calcemia of high producing dairy cows in the first days of lactation, and investigated the impact on the frequency of subclinical hypocalcemia (SCH). Sixty healthy Holstein cows, producing 30 kg of milk/day, handled in intensive system (compost barn), were distributed in groups (n=15) according to lactation order: first, second, third, and fourth to sixth. In the last three weeks before calving they received a diet with negative DCAD (-6mEq/100g DM) and high chloride content. After calving, they received a diet with positive DCAD (18mEq/100g DM). Urine pH was measured before calving. Serum Na+, Cl-, K+, and total Ca concentrations, and the strong ion difference (SID3) were determined in samples taken soon after calving (0h), 24, 48, 72 and 96h after. The frequencies of SCH were determined considering the critical value of 2.125mmol/L (8.5mg/dL). Two-way repeated measures ANOVA and chi-square test were used for comparisons. The cows eliminated acidic urine before calving. Na+, K+, Cl-, and SID3 values did not differ between groups. Na+ and K+ did not vary between days; Cl- was elevated at calving and decreased until 72h; and SID3 was reduced at calving and increased up to 48h. The Ca levels were reduced until 24h and increased up to 72h. Cows of third and fourth to sixth lactations presented lower values up to 24h. SCH was observed in almost half of the cows (43.3% to 55%) until 48h. The maintenance of hypocalcemia for three or more consecutive days occurred in 53.3% of third and fourth to sixth lactations cows. Ingestion of a high chloride prepartum anionic diet led to hyperchloremic acidosis and this imbalance was reversed on the second postpartum day. The induced effects on electrolyte and acid-base balances were not able to prevent the occurrence of SCH in the first days of lactation.(AU)

Os objetivos do estudo foram avaliar os efeitos que a dieta aniônica pré-parto provoca sobre o equilíbrio eletrolítico e sobre a calcemia de vacas leiteiras de alta produção nos primeiros dias de lactação, e verificar o impacto sobre a frequência da hipocalcemia subclínica (HSC). Sessenta fêmeas hígidas HPB, com produção de 30 kg de leite/dia, manejadas em sistema intensivo (compost barn), foram distribuídas por grupos (n=15) de acordo com a ordem de lactação: primeira, segunda, terceira e quarta a sexta. Nas três semanas pré-parto receberam dieta com DCAD negativa (-6mEq/100g MS) e teor de cloreto elevado. Após o parto receberam dieta com DCAD positiva (18mEq/100g MS). O pH da urina foi mensurado antes do parto. As concentrações séricas de Na+, Cl-, K+ e Ca total e a diferença de íons fortes (SID3) foram determinadas em amostras colhidas ao parto (0h), 24, 48, 72 e 96h após. As frequências de HSC foram determinadas considerando-se o valor crítico de 2,125mmol/L (8,5mg/dL). ANOVA de medidas repetidas e teste de qui-quadrado foram empregados para as comparações. As vacas eliminavam urina ácida antes do parto. Os valores de Na+, K+, Cl- e SID3 não diferiram entre os grupos. Na+ e K+ não variaram entre os dias; Cl- era elevado ao parto e diminuiu até 72h; e SID3 era reduzida ao parto e aumentou até 48h. A calcemia era reduzida até 24h e se elevou até 72h. Vacas de terceira e de quarta a sexta lactações apresentaram valores mais baixos até 24h. A HSC foi observada em quase metade das vacas (43,3% a 55%) até 48h. A manutenção de hipocalcemia por três ou mais dias seguidos ocorreu em 53,3% das vacas de terceira e de quarta a sexta lactações. A ingestão de dieta aniônica pré-parto com alto teor de cloreto provocou acidose hiperclorêmica e este desequilíbrio se reverteu no segundo dia pós-parto. Os efeitos induzidos sobre os equilíbrios eletrolítico e ácido base não foram capazes de prevenir a ocorrência de HSC nos primeiros dias da lactação.(AU)

Spatial Variability and Associated Risk Assessment of Nitrate and Ammonium Concentration in Hail Haor Wetland, Northeastern Bangladesh

Nitrate-nitrogen and ammonium-nitrogen concentration were evaluated to assess the current nutrient condition of an ecologically as well as economically important wetland of Bangladesh. The contamination condition of surface water nitrate and ammonium of Hail Haor wetland was assessed to understand its probable risk to human health using the water samples collected from total-fifty monitoring stations, 25 each for two different seasons within 2018 to 2019. Nitrate concentration was measured using the spectrophotometer by colorimetric method, whereas ammonium was quantified using the micro Kjeldahl's distillation method. Statistical and geo-spatial analysis revealed an extensive understanding of the temporal and spatial variability as well as possible source identification of the nutrients in the studied area. Medium to a low level of nitrate ranging from 0.95 up to 9.25 mg/L and high ammonium with values from 0.32 up to 1.92 mg/L was a sporadic trend observed in wetland water, with low water season having more concentration than that of wet high water season. Hazard quotients (HQ) obtained from wetland water for four individual age groups recognized wetland water to be safe for consumption (HQ<1) based on its nitrate level. Wetlands can provide valuable ecosystem services, conserve the native and exotic wetland inhabitants and mitigate flood along with their purifying properties of nutrient removal to lessen the public health concerns about nitrate-nitrogen in drinking water. Therefore, appropriate rein in measures, incentivizing water use efficiency, and continuous monitoring of water quality will ensure safety for wetland habitats, also may aware of the risks or opportunities associated with using wetland water for both drinking and agricultural use.

Glyphosate-resistant soybeans yield in function of glyphosate salts, doses and stadium phenological / Produtuvidade de cultivares de soja em função dos sais de glyphosate, doses e épocas de aplicação

Given the importance of soybean to the global economy and as a food source, improving crop management techniques is integral to obtaining higher yields. As such, this study aimed to assess the yield of soybean cultivars as a function of different glyphosate salt formulations, doses and application times. Two field experiments were conducted simultaneously using a randomized block design and 3x4x2 factorial scheme, with three repetitions. Factor A consisted of three glyphosate formulations: isopropylamine salt (Roundup Original®), ammonium salt (Roundup WG®), and potassium salt (Zapp Qi®), factor B four application times: 0, 14, 28, 42 days after planting (DAP), and factor C two doses: the recommended dose of 720 grams of acid equivalent per hectare and twice that (1440 grams of acid equivalent per hectare). There was no difference between the glyphosate salts for the BMX Ativa RR® cultivar, but yields of BMX Apolo RR® submitted to ammonium salt were 28% higher than those recorded for the other salts studied. Glyphosate potassium salt resulted in lower soybean yields. Potassium salt application between 14 and 28 DAP caused greater toxicity in both the soybean cultivars analyzed.

Diante da importância da cultura da soja tanto na economia mundial quanto para a alimentação, a melhoria das técnicas de cultivo torna-se um ponto fundamental na obtenção de maiores produtividades. Desta forma o objetivo do trabalho foi avaliar a produtividade de cultivares de soja sob a influência da aplicação de diferentes sais de glyphosate, doses e épocas de aplicação. Dois experimentos de campo foram realizados simultaneamente em delineamento de blocos casualizados com arranjo trifatorial (3x4x2) com três repetições. O fator A consistiu de três formulações de glyphosate: sal de isopropilamina (Roundup Original®), sal de amônio (Roundup WG®), sal de potássio (Zapp Qi®). O fator B consistiu de quatro épocas de aplicação: 0, 14, 28, 42 dias após a semeadura (DAS). O fator C consistiu em duas doses: a dose recomendada de 720 gramas de equivalente ácido por hectare e duas vezes a dose recomendada com 1440 gramas de equivalente ácido por hectare. Para a cultivar BMX Ativa RR® não se observou diferença quanto aos sais, porém a cultivar BMX Apolo RR® submetida a aplicação de sal de amônio demostrou uma produtividade 28% superior em relação aos demais sais. O glyphosate na formulação sal de potássio apresentou redução da produtividade da cultura da soja. Aplicações de glyphosate na formulação sal de potássio realizadas entre os 14 e 28 dias após a semeadura causam maior toxidez em ambas cultivares de soja utilizadas.

Kinetic characterization and modeling of a microalgae consortium isolated from landfill leachate under a high CO2 concentration in a bubble column photobioreactor

BACKGROUND: The determination of kinetic parameters and the development of mathematical models are of great interest to predict the growth of microalgae, the consumption of substrate and the design of photobioreactors focused on CO2 capture. However, most of the models in the literature have been developed for CO2 concentrations below 10%. RESULTS: A nonaxenic microalgal consortium was isolated from landfill leachate in order to study its kinetic behavior using a dynamic model. The model considered the CO2 mass transfer from the gas phase to the liquid phase and the effect of light intensity, assimilated nitrogen concentration, ammonium concentration and nitrate concentration. The proposed mathematical model was adjusted with 13 kinetic parameters and validated with a good fit obtained between experimental and simulated data. CONCLUSIONS: Good results were obtained, demonstrating the robustness of the proposed model. The assumption in the model of DIC inhibition in the ammonium and nitrate uptakes was correct, so this aspect should be considered when evaluating the kinetics with microalgae with high inlet CO2 concentrations.